CAJ | 학술논문

背景：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（GalR2）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。
배경：위결혈재관주손상상도치위점막세포개초재、자유기산생과량、백세포침윤、미순배장애。결양후처리능유효감경결양/복양（H/R）조성적손상。감병태수체2（GalR2）주요분포우소화계통화신경계통，대허다내분비활동구유조절작용。목전관우 GalR2대예방위점막상피세포 H/R 손상적작용상미명학。목적：탐토 GalR2격동제후처리대인위점막상피세포 H/R 손상적보호작용급기궤제。방법：이인위점막상피세포 GES-1제비 H/R 손상모형。실험분위정상대조조（N 조）、H/R 조、M1145（GalR2격동제）후처리조（M조）、SB203580（p38MAPK 신호조단제）＋M1145후처리조（S ＋M조）、DMSO 용제대조조（D 조）。이 MTT 검측세포존활솔；류식세포술검측세포조망솔；Hoechst 염색법관찰세포조망정황；ELISA 법검측유산탈경매（LDH）함량；실시정량 PCR 검측 Bcl-2、Bax、p38MAPK 표체수평。결과：H/R 조세포존활솔현저저우 N 조화 M조（P ＜0．05）；H/R 조세포조망솔현저고우 N、M、S ＋M조（P ＜0．05），M조조망솔현저저우 S ＋M조（P ＜0．05）；H/R 조 LDH 함량현저고우 M조화 S ＋M조（P ＜0．05）；N 조、M조 Bcl-2표체수평현저고우 H/R 조、S ＋M조이급 D 조（P ＜0．05）；H/R 조 Bax 표체수평현저고우N、M、S ＋M조（P ＜0．05）；H/R 조、S ＋M조 p38MAPK 표체수평현저저우 M조（P ＜0．05）。결론：GalR2격동제M1145능유효감경 H/R 인기적위점막 GES-1세포손상，차가능통과 p38MAPK 도경발휘작용。
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 post-conditioning 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 +Mgroup)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 significantly 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 Mgroup than that in S +M group (P <0.05).Expression of LDH was significantly higher in H/R group than that in Mand S +Mgroups (P <0.05).Expression of Bcl-2 was significantly higher in N and M groups than that in H/R,S +Mand D groups (P <0.05);expression of Bax was significantly higher in H/R group than that in N,M and S +Mgroups (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.