摘要
目的探讨动力蛋白相关蛋白1(Drp1)抑制剂对肠黏膜上皮细胞缺血再灌注损伤的干预作用并分析其机制。方法将人大肠黏膜上皮细胞系Caco-2分为对照组、模型组、Drp1抑制剂组,对照组正常培养细胞,模型组、Drp1抑制剂组均采用缺氧12 h后复氧2 h的方法构建缺氧复氧模型,Drp1抑制剂组在H/R前给予Drp1抑制剂Mdivi-1干预。采用CCK-8法检测细胞活力,线粒体超氧化物指示剂检测线粒体活性氧(ROS)含量,JC-1法检测线粒体膜电位,流式细胞术检测细胞凋亡率,Western blotting法检测细胞Drp1、线粒体融合蛋白2(Mfn2)蛋白。结果细胞活力对照组>Drp1抑制剂组>模型组(P均<0.05);细胞内线粒体ROS含量模型组>Drp1抑制剂组>对照组,线粒体膜电位对照组>Drp1抑制剂组>模型组(P均<0.05);细胞凋亡率模型组>Drp1抑制剂组>对照组(P均<0.05);细胞Drp1蛋白表达模型组>Drp1抑制剂组>对照组,Mfn2蛋白表达对照组>Drp1抑制剂组>模型组(P均<0.05)。结论Drp1抑制剂可减轻肠黏膜上皮细胞缺血再灌注损伤,其机制可能与改善线粒体功能障碍、减少细胞凋亡有关。
Objective To observe the intervention effect of dynamin-related protein 1(Drp1)inhibitor on ischemiareperfusion injury of intestinal mucosal epithelial cells and to analyze its mechanism.Methods Human colorectal mucosal epithelial cells Caco-2 were divided into the control group,model group,and Drp1 inhibitor group,respectively.The cells in the control group were cultured normally.In the model group and Drp1 inhibitor group,hypoxia-reperfusion models were constructed by using the method of hypoxia for 12 h followed by reoxygenation for 2 h;cells in the Drp1 inhibitor group were given the intervention of the Drp1 inhibitor-Mdivi-1 before the H/R treatment.Cell viability was detected by CCK-8 assay,mitochondrial reactive oxygen species(ROS)content was detected by mitochondrial superoxide indicator,mitochondrial membrane potential level was detected by JC-1 assay,apoptosis rate was detected by flow cytometry,and the expression levels of Drp1,and mitochondrial fusion protein 2(mitofusin2,Mfn2)were detected by Western blotting.Results Cell viability was as follows:control group>Drp1 inhibitor group>model group(all P<0.05),intracellular mitochondrial ROS content was as follows:model group>Drp1 inhibitor group>control group,mitochondrial membrane potential was as follows:control group>Drp1 inhibitor group>model group(all P<0.05),apoptosis rate was as follows:model group>Drp1 inhibitor group>control group(all P<0.05),cellular Drp1 protein expression was as follows:model group>Drp1 inhibitor group>control group,and Mfn2 protein expression was as follows:control group>Drp1 inhibitor group>model group(all P<0.05).Conclusion Drp1 inhibitor could reduce ischemia-reperfusion injury in intestinal mucosal epithelial cells,and its mechanism might be related to improving mitochondrial dysfunction and reducing apoptosis.
作者
图拉妮萨·喀迪尔
张贻帼
景祎馨
廖师师
罗杰
丁可
陈榕
孟庆涛
Tulanisa Kadier;ZHANG Yiguo;JING Yixin;LIAO Shishi;LUO Jie;DING Ke;CHEN Rong;MENG Qingtao(Department of Anesthesiology,Renmin Hospital of Wuhan University,Wuhan 430060,China)
出处
《山东医药》
CAS
2024年第5期31-34,共4页
Shandong Medical Journal
基金
国家自然科学基金资助项目(82172155)
中央高校基本科研业务费专项资金项目(2042022kf1082)
湖北省重点实验室开放项目(2021KEY032)。
作者简介
第一作者:图拉妮萨·喀迪尔(1997-),女,硕士研究生,主要研究方向为MODS机制及防治。E-mail:2021283020275@whu.edu.cn;通信作者:孟庆涛(1976-),男,博士研究生,主任医师,主要研究方向为MODS机制及防治。E-mail:mengqingtao2018@126.com。
