【摘要】 胃缺血再灌注损伤常导致胃黏膜细胞钙超载、自由基产生过量、白细胞浸润、微循环障碍。缺氧后处理能有效减轻缺氧/复氧(H/R)造成的损伤。甘丙肽受体2(GaIR2)主要分布于消化系统和神经系统,对许多内分泌活动具有调节作用。目前关于GalR2对预防胃黏膜上皮细胞H/R损伤的作用尚未明确。目的:探讨GalR2激动剂后处理对人胃黏膜上皮细胞H/R损伤的保护作用及其机制。方法:以人胃黏膜上皮细胞GES-1制备H/R损伤模型。实验分为正常对照组(N组)、H/R组、M1145(GalR2激动剂)后处理组(M组)、SB203580(p38MAPK信号阻断剂)+M1145后处理组(S+M组)、DMSO溶剂对照组(D组)。以MTT检测细胞存活率;流式细胞术检测细胞凋亡率;Hoechst染色法观察细胞凋亡情况;ELISA法检测乳酸脱氢酶(LDH)含量;实时定量PCR检测Bcl-2、Bax、p38MAPK表达水平。结果:H/R组细胞存活率显著低于N组和M组(P<0.05);H/R组细胞凋亡率显著高于N、M、S+M组(P<0.05),M组凋亡率显著低于S+M组(P<0.05);H/R组LDH含量显著高于M组和S+M组(P<0.05);N组、M组Bcl-2表达水平显著高于H/R组、S+M组以及D组(P<0.05);H/R组Bax表达水平显著高于N、M、S+M组(P<0.05);H/R组、S+M组p38MAPK表达水平显著低于M组(P<0.05)。结论:GalR2激动剂M1145能有效减轻H/R引起的胃黏膜GES-1细胞损伤,且可能通过p38MAPK途径发挥作用。更多还原

【Abstract】 Background:Gastric ischemia-reperfusion injury often leads to calcium overload,excessive free radical production,leukocyte infiltration and microcirculation disturbance.Post hypoxic treatment can effectively reduce the injury induced by hypoxia/reoxygenation(H/R).Galanin receptor 2(GalR2) is distributed mainly in the digestive and nervous system,which can regulate many endocrine activity.However,the protective effect of GalR2 on human gastric epithelial cells injury induced by H/R is not clarified.Aims:To investigate the protective effect and its mechanism of GalR2 agonist postconditioning on human gastric epithelial cells injury induced by H/R.Methods;H/R model was constructed on human gastric epithelial cells GES-1.Normal control group(N group),H/R group,M1145(GalR2 agonist) treatment group(M group),SB203580(p38MAPK inhibitor) + M1145 treatment group(S + M group) and DMSO solvent control group(D group) were established.Survival rate of cells was measured by MTT assay.Apoptosis rate of cells was determined by flow cytometry,and cell apoptosis was examined by Hoechst staining.Level of lactate dehydrogenase(LDH) was measured by ELISA.Expressions of Bcl-2,Bax and p38MAPK were determined by real-time quantitative PCR.Results;Survival rate of cells was significantly lower in H/R group than that in N and M groups(P < 0.05).Apoptosis rate of cells was significandy higher in H/R group than that in N,M and S + M groups(P < 0.05),and apoptosis rate of cells was significantly lower in M group than that in S + M group(P <0.05).Expression of LDH was significantly higher in H/R group than that in M and S + M groups(P <0.05).Expression of Bcl-2 was significantly higher in N and M groups than that in H/R,S + M and D groups(P < 0.05);expression of Bax was significandy higher in H/R group than that in N,M and S + M groups(P < 0.05);expression of p38MAPK was significantly lower in H/R and S + M groups than that in M group(P<0.05).Conclusions;GalR2 agonist M1145 plays an effective role in reducing the injury of GES-1 cells induced by H/R,the effect may be conducted through p38MAPK pathway.更多还原