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
AIM： To explore the state of autophagy and related mechanisms in the murine retinal microvascular endothelial cells（RMECs） under hypoxia stimulation.METHODS： The murine RMECs were primarily cultured and randomly d...AIM： To explore the state of autophagy and related mechanisms in the murine retinal microvascular endothelial cells（RMECs） under hypoxia stimulation.METHODS： The murine RMECs were primarily cultured and randomly divided into three groups： hypoxia group（cultured in 1% O_2 environment）, hypoxia＋autophagy inhibition group [pretreated with 5 mmol/L 3-methyladenine（3-MA） for 4 h followed by incubation in 1% O_2] and control group（cultured under normoxic condition）. The state of autophagy in RMECs was examined by assaying the turnover of light chain 3 B（LC3BB） and expression of Beclin-1, Atg3 and Atg5 proteins with Western blotting, by detecting formation of autophagosomes with transmission electron microscopy（TEM） and by counting the number of GFP＋ puncta in RMECs. The protein levels of AMPK, P-AMPK, Akt, P-Akt, m-TOR and P-m TOR were also assayed by Western blotting.RESULTS： Primary murine RMECs were successfully cultured. Under hypoxic conditions, the ratio of LC3BB-Ⅱ/Ⅰ and the expression of Beclin-1, Atg3 and Atg5 proteins were increased when compared with the control group. In addition, the numbers of autophagosome and the GFP＋ puncta were also increased under hypoxia. However, pretreatment with 3-MA obviously attenuated these changes in autophagy in RMECs under hypoxia. Protein expression of P-Akt and P-AMPK was increased but P-m TOR level was decreased in cells exposed to hypoxia. CONCLUSION： In murine RMECs autophagy is activated under hypoxia possibly through activation of the AMPK/mTOR signaling pathway.展开更多
Background:To systematically evaluate the efficacy and safety of traditional Chinese medicine in the treatment of refractory gastroesophageal reflux.Methods:PubMed,The Cochrane Library,Embase,Web of Science,China Nati...Background:To systematically evaluate the efficacy and safety of traditional Chinese medicine in the treatment of refractory gastroesophageal reflux.Methods:PubMed,The Cochrane Library,Embase,Web of Science,China National Knowledge Infrastructure,Wanfang Database,China Science and Technology Journal Database and Chinese BioMedical Literature Database were searched for randomized controlled trials of traditional Chinese medicine in the treatment of refractory gastroesophageal reflux from database establishment time to December 2020.After two researchers independently screened the literature,extracted data and evaluated the bias risk included in the study,RevMan5.3 software was used for meta-analysis.Results:A total of 12 randomized controlled trials,were included,including 893 patients.The results of meta-analysis showed that the total effective rate of the treatment group(relative risk=1.28,95%confidence interval(CI)(1.19,1.38),P<0.00001),RGERDQ(refractory gastroesophageal reflux disease)score(mean difference(MD)=−3.35,95%CI(−4.13,−2.57),P<0.00001],acid reflux(acid in the stomach comes out of the mouth)[MD=−0.30,95%CI(−0.45,−0.15),P<0.00001],heartburn(the feeling that the heart is burned)(MD=−0.44,95%CI(−0.60,−0.29),P<0.00001,and retrosternal pain(MD=−0.27,95%CI(−0.44,−0.10),P<0.00001,belching(MD=−0.40,95%CI(−0.57,−0.24),P<0.00001],endoscopic mucosal score(MD=−0.62,95%CI(−0.78,−0.46),P<0.00001],the differences were statistically significant,and the effective rate of mucosal improvement under endoscopy was P=0.93>0.05,with no statistically significant difference.Conclusion:The current evidence shows that traditional Chinese medicine in the treatment of refractory gastroesophageal reflux disease is better than that of western medicine in the total effective rate,relieving acid reflux,heartburn,retrosternal pain and belching symptoms,but it is impossible to judge the improvement of mucosa under endoscope.Due to the limitations of the quality and quantity of included studies,more high-quality studies are needed to confirm the above conclusions.展开更多
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 the National Natural Science Foundation of China(No.81500726)
文摘AIM： To explore the state of autophagy and related mechanisms in the murine retinal microvascular endothelial cells（RMECs） under hypoxia stimulation.METHODS： The murine RMECs were primarily cultured and randomly divided into three groups： hypoxia group（cultured in 1% O_2 environment）, hypoxia＋autophagy inhibition group [pretreated with 5 mmol/L 3-methyladenine（3-MA） for 4 h followed by incubation in 1% O_2] and control group（cultured under normoxic condition）. The state of autophagy in RMECs was examined by assaying the turnover of light chain 3 B（LC3BB） and expression of Beclin-1, Atg3 and Atg5 proteins with Western blotting, by detecting formation of autophagosomes with transmission electron microscopy（TEM） and by counting the number of GFP＋ puncta in RMECs. The protein levels of AMPK, P-AMPK, Akt, P-Akt, m-TOR and P-m TOR were also assayed by Western blotting.RESULTS： Primary murine RMECs were successfully cultured. Under hypoxic conditions, the ratio of LC3BB-Ⅱ/Ⅰ and the expression of Beclin-1, Atg3 and Atg5 proteins were increased when compared with the control group. In addition, the numbers of autophagosome and the GFP＋ puncta were also increased under hypoxia. However, pretreatment with 3-MA obviously attenuated these changes in autophagy in RMECs under hypoxia. Protein expression of P-Akt and P-AMPK was increased but P-m TOR level was decreased in cells exposed to hypoxia. CONCLUSION： In murine RMECs autophagy is activated under hypoxia possibly through activation of the AMPK/mTOR signaling pathway.
文摘Background:To systematically evaluate the efficacy and safety of traditional Chinese medicine in the treatment of refractory gastroesophageal reflux.Methods:PubMed,The Cochrane Library,Embase,Web of Science,China National Knowledge Infrastructure,Wanfang Database,China Science and Technology Journal Database and Chinese BioMedical Literature Database were searched for randomized controlled trials of traditional Chinese medicine in the treatment of refractory gastroesophageal reflux from database establishment time to December 2020.After two researchers independently screened the literature,extracted data and evaluated the bias risk included in the study,RevMan5.3 software was used for meta-analysis.Results:A total of 12 randomized controlled trials,were included,including 893 patients.The results of meta-analysis showed that the total effective rate of the treatment group(relative risk=1.28,95%confidence interval(CI)(1.19,1.38),P<0.00001),RGERDQ(refractory gastroesophageal reflux disease)score(mean difference(MD)=−3.35,95%CI(−4.13,−2.57),P<0.00001],acid reflux(acid in the stomach comes out of the mouth)[MD=−0.30,95%CI(−0.45,−0.15),P<0.00001],heartburn(the feeling that the heart is burned)(MD=−0.44,95%CI(−0.60,−0.29),P<0.00001,and retrosternal pain(MD=−0.27,95%CI(−0.44,−0.10),P<0.00001,belching(MD=−0.40,95%CI(−0.57,−0.24),P<0.00001],endoscopic mucosal score(MD=−0.62,95%CI(−0.78,−0.46),P<0.00001],the differences were statistically significant,and the effective rate of mucosal improvement under endoscopy was P=0.93>0.05,with no statistically significant difference.Conclusion:The current evidence shows that traditional Chinese medicine in the treatment of refractory gastroesophageal reflux disease is better than that of western medicine in the total effective rate,relieving acid reflux,heartburn,retrosternal pain and belching symptoms,but it is impossible to judge the improvement of mucosa under endoscope.Due to the limitations of the quality and quantity of included studies,more high-quality studies are needed to confirm the above conclusions.
