目的通过体外细胞培养和建立大鼠胆管结扎(b ile duct ligation,BDL)所致阻塞性胆汁淤积动物模型,在蛋白水平观察多耐药相关蛋白MRP3(mu ltidrug resistance-assoc iated prote in 3,MRP3)和核受体RXRα(retinoid-X receptor al-pha,RXR...目的通过体外细胞培养和建立大鼠胆管结扎(b ile duct ligation,BDL)所致阻塞性胆汁淤积动物模型,在蛋白水平观察多耐药相关蛋白MRP3(mu ltidrug resistance-assoc iated prote in 3,MRP3)和核受体RXRα(retinoid-X receptor al-pha,RXRα)表达的关系。方法用鹅去氧胆酸(chenodeoxycholic ac id,CDCA)或苯巴比妥(phenobarb ital,PB)分别刺激培养的肝癌细胞HepG2并建立DBL阻塞性胆汁淤积大鼠模型后,分别抽提HepG2细胞总蛋白、核蛋白和大鼠肝脏细胞膜蛋白和核蛋白,W estern b lot方法检测MRP3和RXRα蛋白表达水平的变化。结果在细胞水平,CDCA和PB可诱导HepG2细胞膜MRP3蛋白表达增高,并抑制细胞核RXRα蛋白表达;在动物体内,BDL大鼠肝脏MRP3明显增加,同时RXRα表达明显下降。结论肝细胞膜蛋白MRP3表达的上调可能与核受体RXRα表达抑制有关。展开更多
目的通过诱导剂刺激体外培养的人肝癌细胞HepG2,观察多耐药相关蛋白MRP3(mu ltidrug resistance-assoc iatedprote in 3,MRP3)和核受体RXRα(retinoid-X receptor alpha,RXRα)蛋白的表达变化。方法用鹅去氧胆酸(chenodeoxycholic ac id...目的通过诱导剂刺激体外培养的人肝癌细胞HepG2,观察多耐药相关蛋白MRP3(mu ltidrug resistance-assoc iatedprote in 3,MRP3)和核受体RXRα(retinoid-X receptor alpha,RXRα)蛋白的表达变化。方法用鹅去氧胆酸(chenodeoxycholic ac id,CDCA)或苯巴比妥(phenobarb ital,PB)分别刺激培养的肝癌细胞HepG2后,抽提HepG2细胞总蛋白和核蛋白,W estern b lot方法检测MRP3和RXRα蛋白表达水平的变化。结果CDCA和PB可诱导HepG2细胞膜MRP3蛋白表达增高,并抑制细胞核受体RXRα蛋白表达。结论HepG2细胞中膜转运蛋白MRP3表达的上调可能与核受体RXRα表达抑制相关。展开更多
OBJECTIVE Neocryptotanshinone(NCTS)is a natural product extracted from traditional Chinese herb Salvia miltiorrhiza Bunge.Previous studies have demonstrated the anti-inflammatory of NCTS in lipopolysaccharide(LPS)-sti...OBJECTIVE Neocryptotanshinone(NCTS)is a natural product extracted from traditional Chinese herb Salvia miltiorrhiza Bunge.Previous studies have demonstrated the anti-inflammatory of NCTS in lipopolysaccharide(LPS)-stimulated mouse macrophage(RAW 264.7).However,the protective effect and mechanism of NCTS in cardiomy⁃ocytes are still undefined.This study is to investigate whether NCTS exerts its cardioprotective effect against hypoxia/re⁃oxygenation(H/R)-induced H9C2 cell injury.METHODS The model of H/R injury was established through hypoxia for 8 h and reoxygenation for 12 h in H9C2 cardiomyocytes of rats.Cultured cardiomyocytes were randomly divided into four groups,control group,H/R group,H/R+NCTS pretreated group(1,2,5 and 10μmol·L^-1),and H/R+NCTS+HX531(an RXRαantagonist,2μmol·L^-1)co-treated group.The cell viability was measured by Cell Counting Kit-8,Hoechst33258 staining was used to observe the morphology of apoptotic changes.Mitochondrial membrane potential was detected by JC-1 fluorescent probe,and protein expressions of RXRα,Bcl-2,Bax,caspase-3 and cleaved caspase-3 with Western blotting.RESULTS Compared with control group,the cell viability in model group was decreased(P<0.05).After treated with NCTS in different concentrations,the CCK8 results showed that NCTS in 2μmol·L^-1 had protective effects.Result of Hoechst33258 staining suggested that the apoptosis was notably increased in model group(P<0.05),Meanwhile,the JC-1 results showed that the mitochondrial membrane potential of the model group decreased which was consistent with previous study.impressively,NCTS could restore the mitochondrial membrane potential as well as apoptosis.Fur⁃ther western blot experiments showed that NCTS treat could upregulate Bcl-2 protein,and downregulate the levels of Bax and cleaved caspase-3/caspase-3 ratio.Since RXRαis a critical upstreaming proteins which can directly mediate the apoptosis,we then determined the effect of NCTS on it.Intriguingly,RXRαwas notably activated by NCTS,while the HX531,the antagonist of RXRα,could abolished NCTS'effect when co-treated with NCTS.CONCLUSION NCTS in 2μmol·L^-1 was effective to protect H9C2 cell from H/R-induced cell injury through RXRα-mediated mitochondria apop⁃tosis.Current results provide possible drugs for the treatment of ischemic cardiomyopathy.展开更多
Myocardial infarction-induced heart failure (HF) is the leading cause of mortality and disability worldwide [1]. Studies have established that HF is characterized by impaired mitochondrial energy production [2]. Hence...Myocardial infarction-induced heart failure (HF) is the leading cause of mortality and disability worldwide [1]. Studies have established that HF is characterized by impaired mitochondrial energy production [2]. Hence, pharmacological targeting of energy production pathways has emerged as a promising strategy against HF [3]. Retinoid X receptor a (RXRa) is a ligand-activated transcription factor.展开更多
文摘目的通过体外细胞培养和建立大鼠胆管结扎(b ile duct ligation,BDL)所致阻塞性胆汁淤积动物模型,在蛋白水平观察多耐药相关蛋白MRP3(mu ltidrug resistance-assoc iated prote in 3,MRP3)和核受体RXRα(retinoid-X receptor al-pha,RXRα)表达的关系。方法用鹅去氧胆酸(chenodeoxycholic ac id,CDCA)或苯巴比妥(phenobarb ital,PB)分别刺激培养的肝癌细胞HepG2并建立DBL阻塞性胆汁淤积大鼠模型后,分别抽提HepG2细胞总蛋白、核蛋白和大鼠肝脏细胞膜蛋白和核蛋白,W estern b lot方法检测MRP3和RXRα蛋白表达水平的变化。结果在细胞水平,CDCA和PB可诱导HepG2细胞膜MRP3蛋白表达增高,并抑制细胞核RXRα蛋白表达;在动物体内,BDL大鼠肝脏MRP3明显增加,同时RXRα表达明显下降。结论肝细胞膜蛋白MRP3表达的上调可能与核受体RXRα表达抑制有关。
文摘目的通过诱导剂刺激体外培养的人肝癌细胞HepG2,观察多耐药相关蛋白MRP3(mu ltidrug resistance-assoc iatedprote in 3,MRP3)和核受体RXRα(retinoid-X receptor alpha,RXRα)蛋白的表达变化。方法用鹅去氧胆酸(chenodeoxycholic ac id,CDCA)或苯巴比妥(phenobarb ital,PB)分别刺激培养的肝癌细胞HepG2后,抽提HepG2细胞总蛋白和核蛋白,W estern b lot方法检测MRP3和RXRα蛋白表达水平的变化。结果CDCA和PB可诱导HepG2细胞膜MRP3蛋白表达增高,并抑制细胞核受体RXRα蛋白表达。结论HepG2细胞中膜转运蛋白MRP3表达的上调可能与核受体RXRα表达抑制相关。
基金National Natural Science Foundation of China(81822049and 81673712)
文摘OBJECTIVE Neocryptotanshinone(NCTS)is a natural product extracted from traditional Chinese herb Salvia miltiorrhiza Bunge.Previous studies have demonstrated the anti-inflammatory of NCTS in lipopolysaccharide(LPS)-stimulated mouse macrophage(RAW 264.7).However,the protective effect and mechanism of NCTS in cardiomy⁃ocytes are still undefined.This study is to investigate whether NCTS exerts its cardioprotective effect against hypoxia/re⁃oxygenation(H/R)-induced H9C2 cell injury.METHODS The model of H/R injury was established through hypoxia for 8 h and reoxygenation for 12 h in H9C2 cardiomyocytes of rats.Cultured cardiomyocytes were randomly divided into four groups,control group,H/R group,H/R+NCTS pretreated group(1,2,5 and 10μmol·L^-1),and H/R+NCTS+HX531(an RXRαantagonist,2μmol·L^-1)co-treated group.The cell viability was measured by Cell Counting Kit-8,Hoechst33258 staining was used to observe the morphology of apoptotic changes.Mitochondrial membrane potential was detected by JC-1 fluorescent probe,and protein expressions of RXRα,Bcl-2,Bax,caspase-3 and cleaved caspase-3 with Western blotting.RESULTS Compared with control group,the cell viability in model group was decreased(P<0.05).After treated with NCTS in different concentrations,the CCK8 results showed that NCTS in 2μmol·L^-1 had protective effects.Result of Hoechst33258 staining suggested that the apoptosis was notably increased in model group(P<0.05),Meanwhile,the JC-1 results showed that the mitochondrial membrane potential of the model group decreased which was consistent with previous study.impressively,NCTS could restore the mitochondrial membrane potential as well as apoptosis.Fur⁃ther western blot experiments showed that NCTS treat could upregulate Bcl-2 protein,and downregulate the levels of Bax and cleaved caspase-3/caspase-3 ratio.Since RXRαis a critical upstreaming proteins which can directly mediate the apoptosis,we then determined the effect of NCTS on it.Intriguingly,RXRαwas notably activated by NCTS,while the HX531,the antagonist of RXRα,could abolished NCTS'effect when co-treated with NCTS.CONCLUSION NCTS in 2μmol·L^-1 was effective to protect H9C2 cell from H/R-induced cell injury through RXRα-mediated mitochondria apop⁃tosis.Current results provide possible drugs for the treatment of ischemic cardiomyopathy.
基金supported by the National Natural Science Foundation of China (82174215, 82230126, 82174364, 82374420,82305025 and 82222075)the Fundamental Research Funds for the Central Universities (Distinguished project)+1 种基金Science and Technology Department of Beijing University of Chinese Medicine(BZY-JMZY-2022-001)“High level Key Discipline of National Administration of Traditional Chinese Medicine-Traditional Chinese Constitutional Medicine (zyyzdxk-2023251)”。
文摘Myocardial infarction-induced heart failure (HF) is the leading cause of mortality and disability worldwide [1]. Studies have established that HF is characterized by impaired mitochondrial energy production [2]. Hence, pharmacological targeting of energy production pathways has emerged as a promising strategy against HF [3]. Retinoid X receptor a (RXRa) is a ligand-activated transcription factor.
