Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-asso...Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.展开更多
Momordica antiviral protein 30 kD(MAP30)is a type I ribosome-inactivating protein(RIP)with antibacterial,anti-HIV and antitumor activities but lacks the ability to target tumor cells.To increase its tumor-targeting ab...Momordica antiviral protein 30 kD(MAP30)is a type I ribosome-inactivating protein(RIP)with antibacterial,anti-HIV and antitumor activities but lacks the ability to target tumor cells.To increase its tumor-targeting ability,the arginine-glycine-aspartic(RGD)peptide and the epidermal growth factor receptor interference(EGFRi)peptide were fused with MAP30,which was named ELRL-MAP30.The efficiency of targeted therapy for triple-negative breast cancer(TNBC)MDA-MB-231 cells,which lack the expression of estrogen receptor(ER),Progesterone receptor(PgR)and human epidermal growth factor receptor-2(HER2),is limited.In this study,we focus on exploring the effect and mechanism of ELRL-MAP30 on TNBC MDA-MB-231 cells.First,we discovered that ELRL-MAP30 significantly inhibited the migration and invasion of MDA-MB-231 cells and induced MDA-MB-231 cell apoptosis.Moreover,ELRL-MAP30 treatment resulted in a significant increase in Bax expression and a decrease in Bcl-2 expression.Furthermore,ELRL-MAP30 triggered apoptosis via the Fak/EGFR/Erk and Ilk/Akt signaling pathways.In addition,recombinant ELRL-MAP30 can inhibit chicken embryonic angiogenesis,and also inhibit the tube formation ability of human umbilical vein endothelial cells(HUVECs),indicating its potential therapeutic effects on tumor angiogenesis.Collectively,these results indicate that ELRL-MAP30 has significant tumor-targeting properties in MDA-MB-231 cancer cells and reveals potential therapeutic effects on angiogenesis.These findings indicate the potential role of ELRL-MAP30 in the targeted treatment of the TNBC cell line MDA-MB-231.展开更多
Ferroptosis, an iron-dependent type of cell death, is being considered for new clinical treatments of malignant tumors that are difficult to treat with apoptosis inducers. Although several reports have attempted to in...Ferroptosis, an iron-dependent type of cell death, is being considered for new clinical treatments of malignant tumors that are difficult to treat with apoptosis inducers. Although several reports have attempted to increase the sensitivity of cells to cell death by combining ferroptosis and apoptosis inducers using a single treatment, detailed elucidation of the respective mechanisms of ferroptosis and apoptosis during cell death remains unclear. Here, we evaluated combined treatment effectiveness using the apoptosis-sensitive rat insulinoma INS-1 cell lines. DNA laddering, an indicator of camptothecin (CPT)-induced apoptosis, was abolished by adding RSL3 and ML-162, but not erastin. We found that when the cells were treated with the apoptosis inducer CPT or the ferroptosis inducer RSL3, respectively, the degree of cytotoxicity observed increased dose-dependently. However, a combined CPT and RSL3 treatment did not show a synergistic decrease in cell viability. Camptothecin did not significantly affect increases in intracellular lipid peroxidation and reactive oxygen species or increases in mitochondrial and cytoplasmic free iron levels that were induced by treatment with RSL3 alone. Moreover, deferoxamine and α-tocopherol were found to inhibit RSL3-induced cytotoxicity but did not protect against CPT or CPT and RSL3-induced cytotoxicity. Finally, the exogenous addition of tert-butyl hydroperoxide inhibited DNA ladder formation that is induced by CPT, while the addition of hydrogen peroxide or ferrous ammonium sulfate had no effect. Taken together, these results suggest that lipid peroxides generated during ferroptosis may suppress cell death induced by apoptotic mechanisms.展开更多
Objective:To investigate the protective effects of Lepidium draba L.(L.draba)on cyclophosphamide(CP)-induced hepatotoxicity and nephrotoxicity in rats.Methods:A total of 36 rats were divided into six groups as follows...Objective:To investigate the protective effects of Lepidium draba L.(L.draba)on cyclophosphamide(CP)-induced hepatotoxicity and nephrotoxicity in rats.Methods:A total of 36 rats were divided into six groups as follows:the sham control group,the CP group(CP 100 mg/kg i.p.on days 1,7,14,21,28,and 35),the CP groups treated with L.draba extract(100,200 and 400 mg/kg of L.draba extract for 28 d),and the L.draba extract alone group(400 mg/kg of L.draba extract for 28 d).Serum parameters of renal and hepatic function,as well as pro-inflammatory and anti-inflammatory cytokines associated with liver and kidney damage were measured.Moreover,Bax,Bcl-2,and caspase-3 gene expression and histopathological changes were assessed.Results:L.draba extract alleviated CP-induced hepatotoxicity and nephrotoxicity by decreasing nitric oxide,TBARS,IL-6,TNF-α,and IL-1βlevels,as well as increasing superoxide dismutase,catalase and glutathione peroxidase activities,and FRAP,MIF,and TGF-βlevels.In addition,the extract downregulated the expression of pro-apoptotic genes(Bax and caspase-3)and mitigated the destruction of glomeruli and renal tubules as well as the degeneration of hepatocytes.Conclusions:L.draba extract can protect hepatic and renal structure and function against CP-induced toxicities,and may be used as a therapeutic agent for CP-induced hepatotoxicity and nephrotoxicity.展开更多
Background Ochratoxin A(OTA)is a toxin widely found in aquafeed ingredients,and hypoxia is a common prob-lem in fish farming.In practice,aquatic animals tend to be more sensitive to hypoxia while feeds are contaminate...Background Ochratoxin A(OTA)is a toxin widely found in aquafeed ingredients,and hypoxia is a common prob-lem in fish farming.In practice,aquatic animals tend to be more sensitive to hypoxia while feeds are contaminated with OTA,but no studies exist in this area.This research investigated the multiple biotoxicities of OTA and hypoxia combined on the liver of grass carp and explored the mitigating effect of curcumin(CUR).Methods A total of 720 healthy juvenile grass carp(11.06±0.05 g)were selected and assigned randomly to 4 experi-mental groups:control group(without OTA and CUR),1.2 mg/kg OTA group,400 mg/kg CUR group,and 1.2 mg/kg OTA+400 mg/kg CUR group with three replicates each for 60 d.Subsequently,32 fish were selected,divided into nor-moxia(18 fish)and hypoxia(18 fish)groups,and subjected to hypoxia stress for 96 h.Results CUR can attenuate histopathological damage caused by coming to OTA and hypoxia by reducing vacu-olation and nuclear excursion.The alleviation of this damage was associated with the attenuation of apoptosis in the mitochondrial pathway by decreasing the expression of the pro-apoptotic proteins Caspase 3,8,9,Bax,and Apaf1 while increasing the expression of the anti-apoptotic protein Bcl-2,and attenuation of endoplasmic reticulum stress(ERS)by reducing Grp78 expression and chop levels.This may be attributed to the fact that the addi-tion of CUR increased the levels of catalase(CAT)and glutathione reductase(GSH),increased antioxidant capacity,and ensured the proper functioning of respiratory chain complexes I and II,which in turn reduced the high produc-tion of reactive oxygen species(ROS),thus alleviating apoptosis and ERS.Conclusions In conclusion,our data demonstrate the effectiveness of CUR in attenuating liver injury caused by the combination of OTA and hypoxia.This study confirms the feasibility and efficacy of adding natural products to mitigate toxic damage to aquatic animals.展开更多
Premature ovarian insufficiency(POI)caused by chemotherapy is a common complication in female cancer survivors of childbearing age.Traditional methods,including mesenchymal stem cell(MSC)transplant and hormone replace...Premature ovarian insufficiency(POI)caused by chemotherapy is a common complication in female cancer survivors of childbearing age.Traditional methods,including mesenchymal stem cell(MSC)transplant and hormone replacement therapy,have limited clinical application because of their drawbacks,and more methods need to be developed.In the current study,the potential effects and underlying mechanisms of human umbilical cord MSC-derived extracellular vesicles(h UCMSC-EVs)were investigated in a cisplatin(CDDP)-induced POI mouse model and a human granulosa cell(GC)line.The results showed that h UCMSC-EVs significantly attenuated body weight loss,ovarian weight loss,ovary atrophy,and follicle loss in moderate-dose(1.5 mg/kg)CDDP-induced POI mice,similar to the effects observed with h UCMSCs.We further found that the h UCMSCEVs inhibited CDDP-induced ovarian GC apoptosis by upregulating anti-apoptotic mi RNA levels in GCs,thereby downregulating the m RNA levels of multiple pro-apoptotic genes.In general,our findings indicate that the moderate-dose chemotherapy may be a better choice for clinical oncotherapy,considering effective rescue of the oncotherapy-induced ovarian damage with h UCMSC-EVs.Additionally,multiple mi RNAs in h UCMSC-EVs may potentially be used to inhibit the chemotherapy-induced ovarian GC apoptosis,thereby restoring ovarian function and improving the life quality of female cancer patients.展开更多
BACKGROUND Acute lung injury(ALI)is a fatal and heterogeneous disease.While bone marrow mesenchymal stem cells(BMSCs)have shown promise in ALI repair,their efficacy is compromised by a high apoptotic percentage.Prelim...BACKGROUND Acute lung injury(ALI)is a fatal and heterogeneous disease.While bone marrow mesenchymal stem cells(BMSCs)have shown promise in ALI repair,their efficacy is compromised by a high apoptotic percentage.Preliminary findings have indicated that long noncoding RNA(lncRNA)-ENST expression is markedly downregulated in MSCs under ischemic and hypoxic conditions,establishing a rationale for in vitro exploration.AIM To elucidate the role of lncRNA-ENST00000517482(lncRNA-ENST)in modulating MSC apoptosis.METHODS Founded on ALI in BEAS-2B cells with lipopolysaccharide,this study employed a transwell co-culture system to study BMSC tropism.BMSCs were genetically modified to overexpress or knockdown lncRNA-ENST.After analyzing the effects on autophagy,apoptosis and cell viability,the lncRNA-ENST/miR-539/c-MYC interaction was confirmed by dual-luciferase assays.RESULTS These findings have revealed a strong correlation between lncRNA-ENST levels and the apoptotic and autophagic status of BMSCs.On the one hand,the overexpression of lncRNA-ENST,as determined by Cell Counting Kit-8 assays,increased the expression of autophagy markers LC3B,ATG7,and ATG5.On the other hand,it reduced apoptosis and boosted BMSC viability.In co-cultures with BEAS-2B cells,lncRNA-ENST overexpression also improved cell vitality.Additionally,by downregulating miR-539 and upregulating c-MYC,lncRNA-ENST was found to influence mitochondrial membrane potential,enhance BMSC autophagy,mitigate apoptosis and lower the secretion of proinflammatory cytokines interleukin-6 and interleukin-1β.Collectively,within the in vitro framework,these results have highlighted the therapeutic potential of BMSCs in ALI and the pivotal regulatory role of lncRNA-ENST in miR-539 and apoptosis in lipopolysaccharide-stimulated BEAS-2B cells.CONCLUSION Our in vitro results show that enhanced lncRNA ENST expression can promote BMSC proliferation and viability by modulating the miR-539/c-MYC axis,reduce apoptosis and induce autophagy,which has suggested its therapeutic potential in the treatment of ALI.展开更多
Objective Emerging evidence suggests that exposure to ultrafine particulate matter(UPM,aerodynamic diameter<0.1μm)is associated with adverse cardiovascular events.Previous studies have found that Shenlian(SL)extra...Objective Emerging evidence suggests that exposure to ultrafine particulate matter(UPM,aerodynamic diameter<0.1μm)is associated with adverse cardiovascular events.Previous studies have found that Shenlian(SL)extract possesses anti-inflammatory and antiapoptotic properties and has a promising protective effect at all stages of the atherosclerotic disease process.In this study,we aimed to investigated whether SL improves UPM-aggravated myocardial ischemic injury by inhibiting inflammation and cell apoptosis.Methods We established a mouse model of MI+UPM.Echocardiographic measurement,measurement of myocardialinfarct size,biochemical analysis,enzyme-linked immunosorbent assay(ELISA),histopathological analysis,Transferase dUTP Nick End Labeling(TUNEL),Western blotting(WB),Polymerase Chain Reaction(PCR)and so on were used to explore the anti-inflammatory and antiapoptotic effects of SL in vivo and in vitro.Results SL treatment can attenuate UPM-induced cardiac dysfunction by improving left ventricular ejection fraction,fractional shortening,and decreasing cardiac infarction area.SL significantly reduced the levels of myocardial enzymes and attenuated UPM-induced morphological alterations.Moreover,SL significantly reduced expression levels of the inflammatory cytokines IL-6,TNF-α,and MCP-1.UPM further increased the infiltration of macrophages in myocardial tissue,whereas SL intervention reversed this phenomenon.UPM also triggered myocardial apoptosis,which was markedly attenuated by SL treatment.The results of in vitro experiments revealed that SL prevented cell damage caused by exposure to UPM combined with hypoxia by reducing the expression of the inflammatory factor NF-κB and inhibiting apoptosis in H9c2 cells.Conclusion Overall,both in vivo and in vitro experiments demonstrated that SL attenuated UPMaggravated myocardial ischemic injury by inhibiting inflammation and cell apoptosis.The mechanisms were related to the downregulation of macrophages infiltrating heart tissues.展开更多
Objectives:Epibrassinolide(EBR)is a steroid hormone with anti-tumor properties.Nevertheless,its potential to inhibit gastric cancer(GC)cells remains unknown.The aim of this research was to examine the effects of EBR o...Objectives:Epibrassinolide(EBR)is a steroid hormone with anti-tumor properties.Nevertheless,its potential to inhibit gastric cancer(GC)cells remains unknown.The aim of this research was to examine the effects of EBR on GC cells and to investigate the specific mechanism of EBR.Methods:A cell counting kit-8(CCK-8)assay was utilized to determine cell survival rates.The investigation of apoptosis,cell cycle progression,and reactive oxygen species(ROS)levels was performed using flow cytometry.To detect cell migration,a wound-healing assay was performed on AGS cells.Furthermore,western blotting assay was utilized to determine protein expression levels.Results:The CCK-8 assay demonstrated that EBR reduced the survival rates of AGS,KATO-3,and MKN-45 cells,while causing only minor toxicity to normal cells.The apoptosis assay indicated that EBR induced AGS cell apoptosis through a mitochondria-mediated pathway.Western blotting results demonstrated that EBR induced AGS cell apoptosis via mitogen-activated protein kinase(MAPK)/signal transducer and activator of transcription 3(STAT3)/nuclear factor kappa B(NF-κB)signaling pathway.Further,after treating AGS cells with EBR,the accumulation of intracellular ROS markedly increased.EBR also induced G2/M phase cell cycle arrest in AGS cells by downregulating phospho-protein kinase B(p-AKT),cyclin-dependent kinase 1/2(CDK1/2),and cyclin B1 expression levels,while simultaneously upregulating p21 and p27 expression levels.EBR inhibited AGS cell migration by downregulating p-AKT,phosphorylated-glycogen synthase kinase 3β(p-GSK-3β),andβ-catenin expression levels and upregulating E-cadherin expression levels.However,these effects were reversed by pretreatment with N-acetylcysteine(NAC).Conclusion:EBR regulates AGS cells by inducing apoptosis and G2/M phase arrest,while also inhibiting cell migration,all of which are mediated through ROS-mediated signaling pathways.Ultimately,these effects suggest a significant role for EBR in regulating cellular processes within AGS cells.展开更多
BACKGROUND Colon cancer is one of the most common malignancies worldwide,and chemo-therapy is a widely used strategy in colon cancer clinical therapy.Chemotherapy resistance is the main cause of recurrence and progres...BACKGROUND Colon cancer is one of the most common malignancies worldwide,and chemo-therapy is a widely used strategy in colon cancer clinical therapy.Chemotherapy resistance is the main cause of recurrence and progression in colon cancer.Thus,novel drugs for treatment are urgently needed.Tetramethylpyrazine(TMP),a component of the traditional Chinese medicine Chuanxiong Hort,has been proven to exhibit a beneficial effect in tumors.AIM To investigate the potential anticancer activity of TMP in colon cancer and the underlying mechanisms.METHODS Colon cancer cells were incubated with different concentrations of TMP.Cell viability was evaluated by crystal violet staining assay,and cell apoptosis was assessed by flow cytometry.Apoptosis-associated protein expression was measured using Western blot analysis.Intracellular reactive oxygen species(ROS)levels were assessed by flow cytometry using DCF fluorescence intensity.Xeno-grafts were established by the subcutaneous injection of colon cancer cells into nude mice;tumor growth was monitored and intracellular ROS was detected in tumors by malondialdehyde assay.RESULTS TMP induced apoptosis of colon cancer cells via the activation of the mitochon-drial pathway.TMP increased the generation of intracellular ROS and triggered mitochondria-mediated apoptosis in a caspase-dependent manner.CONCLUSION Our study demonstrates that TMP induces the apoptosis of colon cancer cells and increases the generation of intracellular ROS.TMP triggers mitochondria-mediated apoptosis in a caspase-dependent manner.The accumu-lation of intracellular ROS is involved in TMP-induced apoptosis.Our findings suggest that TMP may be a potential therapeutic drug for the treatment of colon cancer.展开更多
Objective: To investigate the specific mechanism of hypoxia-inducible factor 1 alpha (HIF-1α) in the regulation of human sperm apoptosis, and to provide a new theoretical reference and scientific basis for the diagno...Objective: To investigate the specific mechanism of hypoxia-inducible factor 1 alpha (HIF-1α) in the regulation of human sperm apoptosis, and to provide a new theoretical reference and scientific basis for the diagnosis and treatment of asthenospermia and other related conditions. Methods: Semen samples were categorized into the normal group and asthenospermia group based on sperm motility criteria. HIF-1α interfering agent cobalt chloride (CoCl2) and guanylate cyclase activator (Lificiguat, YC-1) were added respectively, with a control group established accordingly. Sperm motility (using anterior viability rate as an index), apoptosis level, ATP level, mitochondrial membrane potential, and reactive oxygen species (ROS) level were measured. The expression levels of HIF-1α, p-PI3K, and Bcl-2 in the samples were analyzed using Western blotting. Results: Following CoCl2 treatment, there was a significant increase in sperm apoptosis compared to the normal control group (12.51% ± 2.50% VS 11.15% ± 2.42%);additionally, sperm motility (45.34% ± 3.37% VS 51.36% ± 11.68%), ATP production (11.51 ± 2.87 nM/µL VS 14.99 ± 2.83 nM/µL), ROS levels, and mitochondrial membrane potential all decreased significantly (all P α and p-PI3K increased significantly while Bcl-2 expression decreased (all P α in the YC-1 treatment group were decreased, and the expression level of Bcl-2 was increased (all P α can influence human sperm apoptosis and motility through the PI3K signaling pathway.展开更多
This letter addresses Wang and Zhang's investigation into the role of tankyrase 2(TNKS2)as a pivotal driver of malignancy in non-small cell lung cancer(NSCLC)through mechanisms including apoptosis inhibition,enhan...This letter addresses Wang and Zhang's investigation into the role of tankyrase 2(TNKS2)as a pivotal driver of malignancy in non-small cell lung cancer(NSCLC)through mechanisms including apoptosis inhibition,enhanced cellular migration,andβ-catenin pathway activation.Their study in NSCLC cell lines demonstrates that TNKS2 overexpression stabilizesβ-catenin,subsequently triggering onco-genic gene expression and facilitating cellular migration-key attributes of meta-static potential.These insights position TNKS2 as a compelling target for therapy and a potential prognostic marker in NSCLC.Nevertheless,translating these in vitro findings to clinical practice requires validation in in vivo models.Addi-tionally,further research should investigate TNKS2 expression in patient samples and assess its implications in therapy resistance and combination treatment strategies.展开更多
BACKGROUND Diabetic foot ulcers(DFU)are estimated to affect about 18.6 million people worldwide annually.The pathogenesis of DFU is complex,and the available drugs are not effective.Dl-3-n-butylphthalide(NBP)is a synt...BACKGROUND Diabetic foot ulcers(DFU)are estimated to affect about 18.6 million people worldwide annually.The pathogenesis of DFU is complex,and the available drugs are not effective.Dl-3-n-butylphthalide(NBP)is a synthetic mixture of racemates used in China for the treatment of ischemic stroke.It was initially isolated from the seeds of Apium graveolens Linn,with studies showing its potential role in treating diabetes and its complications.AIM To predict and validate the mechanism by which NBP treats DFU.METHODS Network pharmacological analysis was performed to identify pharmacological targets and signaling pathways mediating the treatment effect of NBP on DFU.In vivo and in vitro experiments were conducted to validate the therapeutic effects and mechanisms of NBP on DFU.RESULTS Network pharmacology analysis identified 26 pharmacological targets of NBP and predicted that NBP could treat DFU partially by modulating apoptosis and vascular signaling pathways.Results from animal experiments showed that NBP significantly improved DFU by increasing neovascularization and fibroblast proliferation.In vitro tests demonstrated that NBP treatment promoted the migration and proliferation of human umbilical vein endothelial cells and human dermal fibroblasts,while inhibiting the apoptosis of human umbilical vein endothelial cells,human dermal fibroblasts,and human keratinocytes cells.CONCLUSION This study found that NBP could treat DFU by decreasing the rate of apoptosis and increasing angiogenesis via the advanced glycation end products-receptor of advanced glycation end products signaling pathway and binding to the heme oxygenase 1,caspase 3,B cell leukemia/lymphoma 2,brain derived neurotrophic factor,and nuclear factor erythroid 2 L2 genes.展开更多
17α-methyltestosterone(17α-MT)is an emerging pollutant,which is harmful to the endocrine system and reproduction of fish.We investigated the effects of different concentrations of 17α-MT(0,5,30,60,and 100 mg/kg)on ...17α-methyltestosterone(17α-MT)is an emerging pollutant,which is harmful to the endocrine system and reproduction of fish.We investigated the effects of different concentrations of 17α-MT(0,5,30,60,and 100 mg/kg)on endoplasmic reticulum stress(ERS)and apoptosis in the liver of Takifugu fasciatus.Results show that:(1)with the increase of 17α-MT treatment concentration,liver transaminases(alanine aminotransferase;aspartate aminotransferase)and the mRNA expression of ERS marker genes(glucose-regulated protein 78;calreticulin)of T.fasciatus were significantly increased compared with the control group(P<0.05);(2)the activity of succinate dehydrogenase(SDH),Caspase3 and Caspase9 in the liver of T.fasciatus increased with the increase of 17α-MT concentration compared with the control group(P<0.05);(3)by using 4-phenylbutyricacid(4-PBA)inhibitors to stimulate ERS through in vitro experiments,the expression of ERS and apoptosis-related genes significantly decreased(P<0.05),and the apoptosis rate of T.fasciatus hepatocytes was significantly inhibited(P<0.05)under 17α-MT treatment.This study confirmed that ERS played an important role in the induction of apoptosis in the hepatocytes of T.fasciatus,which enriched the ecotoxicological information of environmental androgens.展开更多
AIM:To evaluate the role of reactive oxygen speciesendoplasmic reticulum stress(ROS-ERS)in the cellular protection of G protein-coupled receptor 120(GPR120/FFAR4)against high glucose(HG)induced human retinal vascular ...AIM:To evaluate the role of reactive oxygen speciesendoplasmic reticulum stress(ROS-ERS)in the cellular protection of G protein-coupled receptor 120(GPR120/FFAR4)against high glucose(HG)induced human retinal vascular endothelial cell(HRVEC)injury and its underlying mechanisms.METHODS:HRVECs were divided into the control group,GW9508(an agonist of GPR120)group,HG group,and HG+GW9508 group.The cell proliferation and apoptosis were assessed by cell counting kit-8 and annexin V-FITC/PI apoptosis detection kit,respectively.Western blotting analysis was performed to assess the protein expressions of Bax,Bcl-2,activating transcription factor 6(ATF6),PKRlike endoplasmic reticulum kinase(PERK),and inositolrequiring enzyme 1(IRE1).The ROS assay kit was used for the detection of ROS production.Then the cells were transfected with siRNA of GPR120 and the ROS level and protein levels of ATF6,PERK,and IER1 were compared.RESULTS:GW9508 promoted the proliferation of HRVECs,which was significantly reduced by the stimulation of HG.GW9508 remarkably reduced the apoptosis rate of HRVECs under HG and the expression of proapoptotic protein Bax,while increased the expression of antiapoptotic protein Bcl-2.Under HG condition,a significant increase of ROS production was noticed in HRVECs,and GW9508 treatment greatly decreased it.The over-expressions of ERS-related proteins ATF6,PERK,and IER1 under HG were down-regulated by GW9508 treatment.After successfully transfected with siGPR120,the effects of GW9508 on the production of ROS as well as the expressions of ATF6,PERK,and IER1 were reversed.CONCLUSION:GPR120 protects HRVECs against HG induced apoptosis,and suppressing ROS-ERS pathway is one of the mechanisms involved.Activation of GPR120 may be considered as a potential therapeutic target for diabetic retinopathy.展开更多
BACKGROUND Recently,the identification of cell apoptosis induced by natural products has become research hotspot and frontier in the biopharmaceutical and food industries under the umbrella of global green development...BACKGROUND Recently,the identification of cell apoptosis induced by natural products has become research hotspot and frontier in the biopharmaceutical and food industries under the umbrella of global green development worldwide.Traditionally,cell apoptosis is identified using morphological,biochemical,and cell cycle experiments,which is time consuming,and experimental materials are not from the same group,and it is very hard to ensure the identity and veracity of results of former and latter experiments.AIM To establish a selective,instant,and practical protocol to identify cell apoptosis induced by natural products.METHODS A one transient cell processing procedure(OTCPP)was used to detect human colorectal cancer LoVo cell apoptosis after treatment with Pinus massoniana bark extract(PMBE)at the morphological,biochemical,and cell cycle levels.The methods used included treatment with DNA gel electrophoresis,fluorescence microscopy,and flow cytometry.RESULTS In PMBE-treated LoVo cells,we observed a DNA ladder on gel electrophoresis and fluorescence microscopy revealed"nuclear shrinkage,chromatin condensation or fragmentation".In addition,flow cytometry showed an"obvious apoptosis curve".Thus OTCPP achieved synchronous detection of the morphology,biochemistry,cell cycle,and the DNA content of the cells.CONCLUSION OTCPP can quickly identify apoptosis and measure the apoptosis rate,thereby unifying qualitative and quantitative analysis.展开更多
Diabetes mellitus(DM)is a metabolic disorder characterized by persistent hyperglycemia and other symptoms,which pose significant challenges to individual health,life expectancy,and public healthcare systems.The escala...Diabetes mellitus(DM)is a metabolic disorder characterized by persistent hyperglycemia and other symptoms,which pose significant challenges to individual health,life expectancy,and public healthcare systems.The escalating global prevalence of diabetes underscores the need for innovative therapeutic interventions.In this article,we critically comment on the study by Wang et al,published in the World Journal of Diabetes,which elucidates the therapeutic potential of Plantamajoside(PMS)in type 2 DM(T2DM)management.The authors provide evidence for the mechanism of action of PMS in T2DM models,demonstrating prevention of endoplasmic reticulum stress and apoptosis of pancreaticβ-cells via activation of DNAJC1.This manuscript provides a brief review of the pathogenesis of T2DM,explores the various roles of PMS in disease therapy in addition to the DNAJC-related apoptotic and autophagic functions,critically evaluates the experimental approaches employed by Wang et al,and provides recommendations for advancing future research.展开更多
β-Sitosterol is a type of phytosterol that occurs naturally in plants.Previous studies have shown that it has anti-oxidant,anti-hyperlipidemic,anti-inflammatory,immunomodulatory,and anti-tumor effects,but it is unkno...β-Sitosterol is a type of phytosterol that occurs naturally in plants.Previous studies have shown that it has anti-oxidant,anti-hyperlipidemic,anti-inflammatory,immunomodulatory,and anti-tumor effects,but it is unknown whetherβ-sitosterol treatment reduces the effects of ischemic stroke.Here we found that,in a mouse model of ischemic stroke induced by middle cerebral artery occlusion,β-sitosterol reduced the volume of cerebral infarction and brain edema,reduced neuronal apoptosis in brain tissue,and alleviated neurological dysfunction;moreover,β-sitosterol increased the activity of oxygen-and glucose-deprived cerebral cortex neurons and reduced apoptosis.Further investigation showed that the neuroprotective effects ofβ-sitosterol may be related to inhibition of endoplasmic reticulum stress caused by intracellular cholesterol accumulation after ischemic stroke.In addition,β-sitosterol showed high affinity for NPC1L1,a key transporter of cholesterol,and antagonized its activity.In conclusion,β-sitosterol may help treat ischemic stroke by inhibiting neuronal intracellular cholesterol overload/endoplasmic reticulum stress/apoptosis signaling pathways.展开更多
Traumatic brain injury is a severe health problem leading to autophagy and apoptosis in the brain.3,6-Dibromo-beta-fluoro-N-(3-methoxyphenyl)-9H-carbazole-9-propanamine(P7C3-A20)can be neuroprotective in various disea...Traumatic brain injury is a severe health problem leading to autophagy and apoptosis in the brain.3,6-Dibromo-beta-fluoro-N-(3-methoxyphenyl)-9H-carbazole-9-propanamine(P7C3-A20)can be neuroprotective in various diseases,including ischemic stroke and neurodegenerative diseases.However,whether P7C3-A20 has a therapeutic effect on traumatic brain injury and its possible molecular mechanisms are unclear.Therefore,in the present study,we investigated the therapeutic effects of P7C3-A20 on traumatic brain injury and explored the putative underlying molecular mechanisms.We established a traumatic brain injury rat model using a modified weight drop method.P7C3-A20 or vehicle was injected intraperitoneally after traumatic brain injury.Severe neurological deficits were found in rats after traumatic brain injury,with deterioration in balance,walking function,and learning memory.Furthermore,hematoxylin and eosin staining showed significant neuronal cell damage,while terminal deoxynucleotidyl transferase mediated dUTP nick end labeling staining indicated a high rate of apoptosis.The presence of autolysosomes was observed using transmission electron microscope.P7C3-A20 treatment reversed these pathological features.Western blotting showed that P7C3-A20 treatment reduced microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)autophagy protein,apoptosis-related proteins(namely,Bcl-2/adenovirus E1B 19-kDa-interacting protein 3[BNIP3],and Bcl-2 associated x protein[Bax]),and elevated ubiquitin-binding protein p62(p62)autophagy protein expression.Thus,P7C3-A20 can treat traumatic brain injury in rats by inhibiting excessive autophagy and apoptosis.展开更多
In critical care medicine,sepsis is a dangerous systemic condition that is highly prevalent and is associated with high morbidity and mortality rates^([1]).The high mortality rate associated with sepsis is closely rel...In critical care medicine,sepsis is a dangerous systemic condition that is highly prevalent and is associated with high morbidity and mortality rates^([1]).The high mortality rate associated with sepsis is closely related to multi-organ dysfunction,with heart injury being particularly critical and considered the starting point of multi-organ injury^([2]).展开更多
基金supported by the National Natural Science Foundation of China,Nos.82071008(to BL)and 82004001(to XJ)Medical Science and Technology Program of Health Commission of Henan Province,No.LHGJ20210072(to RQ)Science and Technology Department of Henan Province,No.212102310307(to XJ)。
文摘Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.
文摘Momordica antiviral protein 30 kD(MAP30)is a type I ribosome-inactivating protein(RIP)with antibacterial,anti-HIV and antitumor activities but lacks the ability to target tumor cells.To increase its tumor-targeting ability,the arginine-glycine-aspartic(RGD)peptide and the epidermal growth factor receptor interference(EGFRi)peptide were fused with MAP30,which was named ELRL-MAP30.The efficiency of targeted therapy for triple-negative breast cancer(TNBC)MDA-MB-231 cells,which lack the expression of estrogen receptor(ER),Progesterone receptor(PgR)and human epidermal growth factor receptor-2(HER2),is limited.In this study,we focus on exploring the effect and mechanism of ELRL-MAP30 on TNBC MDA-MB-231 cells.First,we discovered that ELRL-MAP30 significantly inhibited the migration and invasion of MDA-MB-231 cells and induced MDA-MB-231 cell apoptosis.Moreover,ELRL-MAP30 treatment resulted in a significant increase in Bax expression and a decrease in Bcl-2 expression.Furthermore,ELRL-MAP30 triggered apoptosis via the Fak/EGFR/Erk and Ilk/Akt signaling pathways.In addition,recombinant ELRL-MAP30 can inhibit chicken embryonic angiogenesis,and also inhibit the tube formation ability of human umbilical vein endothelial cells(HUVECs),indicating its potential therapeutic effects on tumor angiogenesis.Collectively,these results indicate that ELRL-MAP30 has significant tumor-targeting properties in MDA-MB-231 cancer cells and reveals potential therapeutic effects on angiogenesis.These findings indicate the potential role of ELRL-MAP30 in the targeted treatment of the TNBC cell line MDA-MB-231.
文摘Ferroptosis, an iron-dependent type of cell death, is being considered for new clinical treatments of malignant tumors that are difficult to treat with apoptosis inducers. Although several reports have attempted to increase the sensitivity of cells to cell death by combining ferroptosis and apoptosis inducers using a single treatment, detailed elucidation of the respective mechanisms of ferroptosis and apoptosis during cell death remains unclear. Here, we evaluated combined treatment effectiveness using the apoptosis-sensitive rat insulinoma INS-1 cell lines. DNA laddering, an indicator of camptothecin (CPT)-induced apoptosis, was abolished by adding RSL3 and ML-162, but not erastin. We found that when the cells were treated with the apoptosis inducer CPT or the ferroptosis inducer RSL3, respectively, the degree of cytotoxicity observed increased dose-dependently. However, a combined CPT and RSL3 treatment did not show a synergistic decrease in cell viability. Camptothecin did not significantly affect increases in intracellular lipid peroxidation and reactive oxygen species or increases in mitochondrial and cytoplasmic free iron levels that were induced by treatment with RSL3 alone. Moreover, deferoxamine and α-tocopherol were found to inhibit RSL3-induced cytotoxicity but did not protect against CPT or CPT and RSL3-induced cytotoxicity. Finally, the exogenous addition of tert-butyl hydroperoxide inhibited DNA ladder formation that is induced by CPT, while the addition of hydrogen peroxide or ferrous ammonium sulfate had no effect. Taken together, these results suggest that lipid peroxides generated during ferroptosis may suppress cell death induced by apoptotic mechanisms.
基金supported by the Basic Research Joint Special General Project of Yunnan Provincial Local Universities(part)(No:202301BA070001-029,202301BA070001-044)Yunnan Province High-level Scientific and Technological Talents and Innovation Team Selection Special Young and Middle-aged Academic and Technical Leaders Reserve Talent Project(No:202405AC350067).
文摘Objective:To investigate the protective effects of Lepidium draba L.(L.draba)on cyclophosphamide(CP)-induced hepatotoxicity and nephrotoxicity in rats.Methods:A total of 36 rats were divided into six groups as follows:the sham control group,the CP group(CP 100 mg/kg i.p.on days 1,7,14,21,28,and 35),the CP groups treated with L.draba extract(100,200 and 400 mg/kg of L.draba extract for 28 d),and the L.draba extract alone group(400 mg/kg of L.draba extract for 28 d).Serum parameters of renal and hepatic function,as well as pro-inflammatory and anti-inflammatory cytokines associated with liver and kidney damage were measured.Moreover,Bax,Bcl-2,and caspase-3 gene expression and histopathological changes were assessed.Results:L.draba extract alleviated CP-induced hepatotoxicity and nephrotoxicity by decreasing nitric oxide,TBARS,IL-6,TNF-α,and IL-1βlevels,as well as increasing superoxide dismutase,catalase and glutathione peroxidase activities,and FRAP,MIF,and TGF-βlevels.In addition,the extract downregulated the expression of pro-apoptotic genes(Bax and caspase-3)and mitigated the destruction of glomeruli and renal tubules as well as the degeneration of hepatocytes.Conclusions:L.draba extract can protect hepatic and renal structure and function against CP-induced toxicities,and may be used as a therapeutic agent for CP-induced hepatotoxicity and nephrotoxicity.
基金This research was financially supported by the earmarked fund for CARS(CARS-45)National Natural Science Foundation of China(32273144,32072985)National Key R&D Program of China(2019YFD0900200).
文摘Background Ochratoxin A(OTA)is a toxin widely found in aquafeed ingredients,and hypoxia is a common prob-lem in fish farming.In practice,aquatic animals tend to be more sensitive to hypoxia while feeds are contaminated with OTA,but no studies exist in this area.This research investigated the multiple biotoxicities of OTA and hypoxia combined on the liver of grass carp and explored the mitigating effect of curcumin(CUR).Methods A total of 720 healthy juvenile grass carp(11.06±0.05 g)were selected and assigned randomly to 4 experi-mental groups:control group(without OTA and CUR),1.2 mg/kg OTA group,400 mg/kg CUR group,and 1.2 mg/kg OTA+400 mg/kg CUR group with three replicates each for 60 d.Subsequently,32 fish were selected,divided into nor-moxia(18 fish)and hypoxia(18 fish)groups,and subjected to hypoxia stress for 96 h.Results CUR can attenuate histopathological damage caused by coming to OTA and hypoxia by reducing vacu-olation and nuclear excursion.The alleviation of this damage was associated with the attenuation of apoptosis in the mitochondrial pathway by decreasing the expression of the pro-apoptotic proteins Caspase 3,8,9,Bax,and Apaf1 while increasing the expression of the anti-apoptotic protein Bcl-2,and attenuation of endoplasmic reticulum stress(ERS)by reducing Grp78 expression and chop levels.This may be attributed to the fact that the addi-tion of CUR increased the levels of catalase(CAT)and glutathione reductase(GSH),increased antioxidant capacity,and ensured the proper functioning of respiratory chain complexes I and II,which in turn reduced the high produc-tion of reactive oxygen species(ROS),thus alleviating apoptosis and ERS.Conclusions In conclusion,our data demonstrate the effectiveness of CUR in attenuating liver injury caused by the combination of OTA and hypoxia.This study confirms the feasibility and efficacy of adding natural products to mitigate toxic damage to aquatic animals.
基金Financial support for the current study was provided by the grant from"Blue Engineering"Excellent Young Teacher Foundation in Colleges and Universities of Jiangsu Province(Grant No.KY101R202207 to Xiaojun Chen)。
文摘Premature ovarian insufficiency(POI)caused by chemotherapy is a common complication in female cancer survivors of childbearing age.Traditional methods,including mesenchymal stem cell(MSC)transplant and hormone replacement therapy,have limited clinical application because of their drawbacks,and more methods need to be developed.In the current study,the potential effects and underlying mechanisms of human umbilical cord MSC-derived extracellular vesicles(h UCMSC-EVs)were investigated in a cisplatin(CDDP)-induced POI mouse model and a human granulosa cell(GC)line.The results showed that h UCMSC-EVs significantly attenuated body weight loss,ovarian weight loss,ovary atrophy,and follicle loss in moderate-dose(1.5 mg/kg)CDDP-induced POI mice,similar to the effects observed with h UCMSCs.We further found that the h UCMSCEVs inhibited CDDP-induced ovarian GC apoptosis by upregulating anti-apoptotic mi RNA levels in GCs,thereby downregulating the m RNA levels of multiple pro-apoptotic genes.In general,our findings indicate that the moderate-dose chemotherapy may be a better choice for clinical oncotherapy,considering effective rescue of the oncotherapy-induced ovarian damage with h UCMSC-EVs.Additionally,multiple mi RNAs in h UCMSC-EVs may potentially be used to inhibit the chemotherapy-induced ovarian GC apoptosis,thereby restoring ovarian function and improving the life quality of female cancer patients.
基金Supported by the Peak Supporting Clinical Discipline of Shanghai Health Bureau,No.2023ZDFC0104the National Key R&D Program of China,No.2019YFA0110601.
文摘BACKGROUND Acute lung injury(ALI)is a fatal and heterogeneous disease.While bone marrow mesenchymal stem cells(BMSCs)have shown promise in ALI repair,their efficacy is compromised by a high apoptotic percentage.Preliminary findings have indicated that long noncoding RNA(lncRNA)-ENST expression is markedly downregulated in MSCs under ischemic and hypoxic conditions,establishing a rationale for in vitro exploration.AIM To elucidate the role of lncRNA-ENST00000517482(lncRNA-ENST)in modulating MSC apoptosis.METHODS Founded on ALI in BEAS-2B cells with lipopolysaccharide,this study employed a transwell co-culture system to study BMSC tropism.BMSCs were genetically modified to overexpress or knockdown lncRNA-ENST.After analyzing the effects on autophagy,apoptosis and cell viability,the lncRNA-ENST/miR-539/c-MYC interaction was confirmed by dual-luciferase assays.RESULTS These findings have revealed a strong correlation between lncRNA-ENST levels and the apoptotic and autophagic status of BMSCs.On the one hand,the overexpression of lncRNA-ENST,as determined by Cell Counting Kit-8 assays,increased the expression of autophagy markers LC3B,ATG7,and ATG5.On the other hand,it reduced apoptosis and boosted BMSC viability.In co-cultures with BEAS-2B cells,lncRNA-ENST overexpression also improved cell vitality.Additionally,by downregulating miR-539 and upregulating c-MYC,lncRNA-ENST was found to influence mitochondrial membrane potential,enhance BMSC autophagy,mitigate apoptosis and lower the secretion of proinflammatory cytokines interleukin-6 and interleukin-1β.Collectively,within the in vitro framework,these results have highlighted the therapeutic potential of BMSCs in ALI and the pivotal regulatory role of lncRNA-ENST in miR-539 and apoptosis in lipopolysaccharide-stimulated BEAS-2B cells.CONCLUSION Our in vitro results show that enhanced lncRNA ENST expression can promote BMSC proliferation and viability by modulating the miR-539/c-MYC axis,reduce apoptosis and induce autophagy,which has suggested its therapeutic potential in the treatment of ALI.
基金supported by CACMS Innovation Fund(No CI2021A04611,CI2021A05106)Scientific and technological innovation project of China Academy of Chinese Medical Sciences(CI2021B015)+1 种基金Scientific and technological innovation project of China Academy of Chinese Medical Sciences(CI2023E001TS01)Fundamental research funds for the central public welfare research institutes(L2022035).
文摘Objective Emerging evidence suggests that exposure to ultrafine particulate matter(UPM,aerodynamic diameter<0.1μm)is associated with adverse cardiovascular events.Previous studies have found that Shenlian(SL)extract possesses anti-inflammatory and antiapoptotic properties and has a promising protective effect at all stages of the atherosclerotic disease process.In this study,we aimed to investigated whether SL improves UPM-aggravated myocardial ischemic injury by inhibiting inflammation and cell apoptosis.Methods We established a mouse model of MI+UPM.Echocardiographic measurement,measurement of myocardialinfarct size,biochemical analysis,enzyme-linked immunosorbent assay(ELISA),histopathological analysis,Transferase dUTP Nick End Labeling(TUNEL),Western blotting(WB),Polymerase Chain Reaction(PCR)and so on were used to explore the anti-inflammatory and antiapoptotic effects of SL in vivo and in vitro.Results SL treatment can attenuate UPM-induced cardiac dysfunction by improving left ventricular ejection fraction,fractional shortening,and decreasing cardiac infarction area.SL significantly reduced the levels of myocardial enzymes and attenuated UPM-induced morphological alterations.Moreover,SL significantly reduced expression levels of the inflammatory cytokines IL-6,TNF-α,and MCP-1.UPM further increased the infiltration of macrophages in myocardial tissue,whereas SL intervention reversed this phenomenon.UPM also triggered myocardial apoptosis,which was markedly attenuated by SL treatment.The results of in vitro experiments revealed that SL prevented cell damage caused by exposure to UPM combined with hypoxia by reducing the expression of the inflammatory factor NF-κB and inhibiting apoptosis in H9c2 cells.Conclusion Overall,both in vivo and in vitro experiments demonstrated that SL attenuated UPMaggravated myocardial ischemic injury by inhibiting inflammation and cell apoptosis.The mechanisms were related to the downregulation of macrophages infiltrating heart tissues.
基金supported by the Heilongjiang Province Key Research and Development Plan Guidance Project[Grant No.GZ20220039]the Central Government Supports the Local College Reform and Development Fund Talent Training Project[Grant No.2020GSP16].
文摘Objectives:Epibrassinolide(EBR)is a steroid hormone with anti-tumor properties.Nevertheless,its potential to inhibit gastric cancer(GC)cells remains unknown.The aim of this research was to examine the effects of EBR on GC cells and to investigate the specific mechanism of EBR.Methods:A cell counting kit-8(CCK-8)assay was utilized to determine cell survival rates.The investigation of apoptosis,cell cycle progression,and reactive oxygen species(ROS)levels was performed using flow cytometry.To detect cell migration,a wound-healing assay was performed on AGS cells.Furthermore,western blotting assay was utilized to determine protein expression levels.Results:The CCK-8 assay demonstrated that EBR reduced the survival rates of AGS,KATO-3,and MKN-45 cells,while causing only minor toxicity to normal cells.The apoptosis assay indicated that EBR induced AGS cell apoptosis through a mitochondria-mediated pathway.Western blotting results demonstrated that EBR induced AGS cell apoptosis via mitogen-activated protein kinase(MAPK)/signal transducer and activator of transcription 3(STAT3)/nuclear factor kappa B(NF-κB)signaling pathway.Further,after treating AGS cells with EBR,the accumulation of intracellular ROS markedly increased.EBR also induced G2/M phase cell cycle arrest in AGS cells by downregulating phospho-protein kinase B(p-AKT),cyclin-dependent kinase 1/2(CDK1/2),and cyclin B1 expression levels,while simultaneously upregulating p21 and p27 expression levels.EBR inhibited AGS cell migration by downregulating p-AKT,phosphorylated-glycogen synthase kinase 3β(p-GSK-3β),andβ-catenin expression levels and upregulating E-cadherin expression levels.However,these effects were reversed by pretreatment with N-acetylcysteine(NAC).Conclusion:EBR regulates AGS cells by inducing apoptosis and G2/M phase arrest,while also inhibiting cell migration,all of which are mediated through ROS-mediated signaling pathways.Ultimately,these effects suggest a significant role for EBR in regulating cellular processes within AGS cells.
文摘BACKGROUND Colon cancer is one of the most common malignancies worldwide,and chemo-therapy is a widely used strategy in colon cancer clinical therapy.Chemotherapy resistance is the main cause of recurrence and progression in colon cancer.Thus,novel drugs for treatment are urgently needed.Tetramethylpyrazine(TMP),a component of the traditional Chinese medicine Chuanxiong Hort,has been proven to exhibit a beneficial effect in tumors.AIM To investigate the potential anticancer activity of TMP in colon cancer and the underlying mechanisms.METHODS Colon cancer cells were incubated with different concentrations of TMP.Cell viability was evaluated by crystal violet staining assay,and cell apoptosis was assessed by flow cytometry.Apoptosis-associated protein expression was measured using Western blot analysis.Intracellular reactive oxygen species(ROS)levels were assessed by flow cytometry using DCF fluorescence intensity.Xeno-grafts were established by the subcutaneous injection of colon cancer cells into nude mice;tumor growth was monitored and intracellular ROS was detected in tumors by malondialdehyde assay.RESULTS TMP induced apoptosis of colon cancer cells via the activation of the mitochon-drial pathway.TMP increased the generation of intracellular ROS and triggered mitochondria-mediated apoptosis in a caspase-dependent manner.CONCLUSION Our study demonstrates that TMP induces the apoptosis of colon cancer cells and increases the generation of intracellular ROS.TMP triggers mitochondria-mediated apoptosis in a caspase-dependent manner.The accumu-lation of intracellular ROS is involved in TMP-induced apoptosis.Our findings suggest that TMP may be a potential therapeutic drug for the treatment of colon cancer.
文摘Objective: To investigate the specific mechanism of hypoxia-inducible factor 1 alpha (HIF-1α) in the regulation of human sperm apoptosis, and to provide a new theoretical reference and scientific basis for the diagnosis and treatment of asthenospermia and other related conditions. Methods: Semen samples were categorized into the normal group and asthenospermia group based on sperm motility criteria. HIF-1α interfering agent cobalt chloride (CoCl2) and guanylate cyclase activator (Lificiguat, YC-1) were added respectively, with a control group established accordingly. Sperm motility (using anterior viability rate as an index), apoptosis level, ATP level, mitochondrial membrane potential, and reactive oxygen species (ROS) level were measured. The expression levels of HIF-1α, p-PI3K, and Bcl-2 in the samples were analyzed using Western blotting. Results: Following CoCl2 treatment, there was a significant increase in sperm apoptosis compared to the normal control group (12.51% ± 2.50% VS 11.15% ± 2.42%);additionally, sperm motility (45.34% ± 3.37% VS 51.36% ± 11.68%), ATP production (11.51 ± 2.87 nM/µL VS 14.99 ± 2.83 nM/µL), ROS levels, and mitochondrial membrane potential all decreased significantly (all P α and p-PI3K increased significantly while Bcl-2 expression decreased (all P α in the YC-1 treatment group were decreased, and the expression level of Bcl-2 was increased (all P α can influence human sperm apoptosis and motility through the PI3K signaling pathway.
文摘This letter addresses Wang and Zhang's investigation into the role of tankyrase 2(TNKS2)as a pivotal driver of malignancy in non-small cell lung cancer(NSCLC)through mechanisms including apoptosis inhibition,enhanced cellular migration,andβ-catenin pathway activation.Their study in NSCLC cell lines demonstrates that TNKS2 overexpression stabilizesβ-catenin,subsequently triggering onco-genic gene expression and facilitating cellular migration-key attributes of meta-static potential.These insights position TNKS2 as a compelling target for therapy and a potential prognostic marker in NSCLC.Nevertheless,translating these in vitro findings to clinical practice requires validation in in vivo models.Addi-tionally,further research should investigate TNKS2 expression in patient samples and assess its implications in therapy resistance and combination treatment strategies.
基金Supported by General Project of Xuzhou Science and Technology Bureau,No.KC22070.
文摘BACKGROUND Diabetic foot ulcers(DFU)are estimated to affect about 18.6 million people worldwide annually.The pathogenesis of DFU is complex,and the available drugs are not effective.Dl-3-n-butylphthalide(NBP)is a synthetic mixture of racemates used in China for the treatment of ischemic stroke.It was initially isolated from the seeds of Apium graveolens Linn,with studies showing its potential role in treating diabetes and its complications.AIM To predict and validate the mechanism by which NBP treats DFU.METHODS Network pharmacological analysis was performed to identify pharmacological targets and signaling pathways mediating the treatment effect of NBP on DFU.In vivo and in vitro experiments were conducted to validate the therapeutic effects and mechanisms of NBP on DFU.RESULTS Network pharmacology analysis identified 26 pharmacological targets of NBP and predicted that NBP could treat DFU partially by modulating apoptosis and vascular signaling pathways.Results from animal experiments showed that NBP significantly improved DFU by increasing neovascularization and fibroblast proliferation.In vitro tests demonstrated that NBP treatment promoted the migration and proliferation of human umbilical vein endothelial cells and human dermal fibroblasts,while inhibiting the apoptosis of human umbilical vein endothelial cells,human dermal fibroblasts,and human keratinocytes cells.CONCLUSION This study found that NBP could treat DFU by decreasing the rate of apoptosis and increasing angiogenesis via the advanced glycation end products-receptor of advanced glycation end products signaling pathway and binding to the heme oxygenase 1,caspase 3,B cell leukemia/lymphoma 2,brain derived neurotrophic factor,and nuclear factor erythroid 2 L2 genes.
基金Supported by the Jiangsu Agriculture Science and Technology Innovation Fund(No.CX(22)2029)the National Natural Science Foundation of China(Nos.32172948,31800436)+1 种基金the“JBGS”Project of Seed Industry Revitalization in Jiangsu Province(No.JBGS(2021)034)the Postgraduate Research&Practice Innovation Program of Jiangsu Province(No.SJCX23_0616)。
文摘17α-methyltestosterone(17α-MT)is an emerging pollutant,which is harmful to the endocrine system and reproduction of fish.We investigated the effects of different concentrations of 17α-MT(0,5,30,60,and 100 mg/kg)on endoplasmic reticulum stress(ERS)and apoptosis in the liver of Takifugu fasciatus.Results show that:(1)with the increase of 17α-MT treatment concentration,liver transaminases(alanine aminotransferase;aspartate aminotransferase)and the mRNA expression of ERS marker genes(glucose-regulated protein 78;calreticulin)of T.fasciatus were significantly increased compared with the control group(P<0.05);(2)the activity of succinate dehydrogenase(SDH),Caspase3 and Caspase9 in the liver of T.fasciatus increased with the increase of 17α-MT concentration compared with the control group(P<0.05);(3)by using 4-phenylbutyricacid(4-PBA)inhibitors to stimulate ERS through in vitro experiments,the expression of ERS and apoptosis-related genes significantly decreased(P<0.05),and the apoptosis rate of T.fasciatus hepatocytes was significantly inhibited(P<0.05)under 17α-MT treatment.This study confirmed that ERS played an important role in the induction of apoptosis in the hepatocytes of T.fasciatus,which enriched the ecotoxicological information of environmental androgens.
文摘AIM:To evaluate the role of reactive oxygen speciesendoplasmic reticulum stress(ROS-ERS)in the cellular protection of G protein-coupled receptor 120(GPR120/FFAR4)against high glucose(HG)induced human retinal vascular endothelial cell(HRVEC)injury and its underlying mechanisms.METHODS:HRVECs were divided into the control group,GW9508(an agonist of GPR120)group,HG group,and HG+GW9508 group.The cell proliferation and apoptosis were assessed by cell counting kit-8 and annexin V-FITC/PI apoptosis detection kit,respectively.Western blotting analysis was performed to assess the protein expressions of Bax,Bcl-2,activating transcription factor 6(ATF6),PKRlike endoplasmic reticulum kinase(PERK),and inositolrequiring enzyme 1(IRE1).The ROS assay kit was used for the detection of ROS production.Then the cells were transfected with siRNA of GPR120 and the ROS level and protein levels of ATF6,PERK,and IER1 were compared.RESULTS:GW9508 promoted the proliferation of HRVECs,which was significantly reduced by the stimulation of HG.GW9508 remarkably reduced the apoptosis rate of HRVECs under HG and the expression of proapoptotic protein Bax,while increased the expression of antiapoptotic protein Bcl-2.Under HG condition,a significant increase of ROS production was noticed in HRVECs,and GW9508 treatment greatly decreased it.The over-expressions of ERS-related proteins ATF6,PERK,and IER1 under HG were down-regulated by GW9508 treatment.After successfully transfected with siGPR120,the effects of GW9508 on the production of ROS as well as the expressions of ATF6,PERK,and IER1 were reversed.CONCLUSION:GPR120 protects HRVECs against HG induced apoptosis,and suppressing ROS-ERS pathway is one of the mechanisms involved.Activation of GPR120 may be considered as a potential therapeutic target for diabetic retinopathy.
基金Supported by 2023 Key R&D Projects of Tongji University,No.150029607160-24323Industrial Key Program Foundation of Guangdong Province:R&D of Natural Novel Anticancer Drugs,No.2004B10401033。
文摘BACKGROUND Recently,the identification of cell apoptosis induced by natural products has become research hotspot and frontier in the biopharmaceutical and food industries under the umbrella of global green development worldwide.Traditionally,cell apoptosis is identified using morphological,biochemical,and cell cycle experiments,which is time consuming,and experimental materials are not from the same group,and it is very hard to ensure the identity and veracity of results of former and latter experiments.AIM To establish a selective,instant,and practical protocol to identify cell apoptosis induced by natural products.METHODS A one transient cell processing procedure(OTCPP)was used to detect human colorectal cancer LoVo cell apoptosis after treatment with Pinus massoniana bark extract(PMBE)at the morphological,biochemical,and cell cycle levels.The methods used included treatment with DNA gel electrophoresis,fluorescence microscopy,and flow cytometry.RESULTS In PMBE-treated LoVo cells,we observed a DNA ladder on gel electrophoresis and fluorescence microscopy revealed"nuclear shrinkage,chromatin condensation or fragmentation".In addition,flow cytometry showed an"obvious apoptosis curve".Thus OTCPP achieved synchronous detection of the morphology,biochemistry,cell cycle,and the DNA content of the cells.CONCLUSION OTCPP can quickly identify apoptosis and measure the apoptosis rate,thereby unifying qualitative and quantitative analysis.
文摘Diabetes mellitus(DM)is a metabolic disorder characterized by persistent hyperglycemia and other symptoms,which pose significant challenges to individual health,life expectancy,and public healthcare systems.The escalating global prevalence of diabetes underscores the need for innovative therapeutic interventions.In this article,we critically comment on the study by Wang et al,published in the World Journal of Diabetes,which elucidates the therapeutic potential of Plantamajoside(PMS)in type 2 DM(T2DM)management.The authors provide evidence for the mechanism of action of PMS in T2DM models,demonstrating prevention of endoplasmic reticulum stress and apoptosis of pancreaticβ-cells via activation of DNAJC1.This manuscript provides a brief review of the pathogenesis of T2DM,explores the various roles of PMS in disease therapy in addition to the DNAJC-related apoptotic and autophagic functions,critically evaluates the experimental approaches employed by Wang et al,and provides recommendations for advancing future research.
基金supported by the National Natural Science Foundation of China,Nos.82104158(to XT),31800887(to LY),31972902(to LY),82001422(to YL)China Postdoctoral Science Foundation,No.2020M683750(to LY)partially by Young Talent Fund of University Association for Science and Technology in Shaanxi Province of China,No.20200307(to LY).
文摘β-Sitosterol is a type of phytosterol that occurs naturally in plants.Previous studies have shown that it has anti-oxidant,anti-hyperlipidemic,anti-inflammatory,immunomodulatory,and anti-tumor effects,but it is unknown whetherβ-sitosterol treatment reduces the effects of ischemic stroke.Here we found that,in a mouse model of ischemic stroke induced by middle cerebral artery occlusion,β-sitosterol reduced the volume of cerebral infarction and brain edema,reduced neuronal apoptosis in brain tissue,and alleviated neurological dysfunction;moreover,β-sitosterol increased the activity of oxygen-and glucose-deprived cerebral cortex neurons and reduced apoptosis.Further investigation showed that the neuroprotective effects ofβ-sitosterol may be related to inhibition of endoplasmic reticulum stress caused by intracellular cholesterol accumulation after ischemic stroke.In addition,β-sitosterol showed high affinity for NPC1L1,a key transporter of cholesterol,and antagonized its activity.In conclusion,β-sitosterol may help treat ischemic stroke by inhibiting neuronal intracellular cholesterol overload/endoplasmic reticulum stress/apoptosis signaling pathways.
基金supported by National Natural Science Foundation of China,No.32102745(to XL).
文摘Traumatic brain injury is a severe health problem leading to autophagy and apoptosis in the brain.3,6-Dibromo-beta-fluoro-N-(3-methoxyphenyl)-9H-carbazole-9-propanamine(P7C3-A20)can be neuroprotective in various diseases,including ischemic stroke and neurodegenerative diseases.However,whether P7C3-A20 has a therapeutic effect on traumatic brain injury and its possible molecular mechanisms are unclear.Therefore,in the present study,we investigated the therapeutic effects of P7C3-A20 on traumatic brain injury and explored the putative underlying molecular mechanisms.We established a traumatic brain injury rat model using a modified weight drop method.P7C3-A20 or vehicle was injected intraperitoneally after traumatic brain injury.Severe neurological deficits were found in rats after traumatic brain injury,with deterioration in balance,walking function,and learning memory.Furthermore,hematoxylin and eosin staining showed significant neuronal cell damage,while terminal deoxynucleotidyl transferase mediated dUTP nick end labeling staining indicated a high rate of apoptosis.The presence of autolysosomes was observed using transmission electron microscope.P7C3-A20 treatment reversed these pathological features.Western blotting showed that P7C3-A20 treatment reduced microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)autophagy protein,apoptosis-related proteins(namely,Bcl-2/adenovirus E1B 19-kDa-interacting protein 3[BNIP3],and Bcl-2 associated x protein[Bax]),and elevated ubiquitin-binding protein p62(p62)autophagy protein expression.Thus,P7C3-A20 can treat traumatic brain injury in rats by inhibiting excessive autophagy and apoptosis.
基金supported by Jiangsu Traditional Chinese Medicine Science and Technology Development Program(MS2022099)The Postgraduate Research&Practice Innovation Program of Jiangsu Ocean University(No.KYCX2022-34)。
文摘In critical care medicine,sepsis is a dangerous systemic condition that is highly prevalent and is associated with high morbidity and mortality rates^([1]).The high mortality rate associated with sepsis is closely related to multi-organ dysfunction,with heart injury being particularly critical and considered the starting point of multi-organ injury^([2]).
