SIRT5通过促进IDH2去琥珀酰化增加COPD小鼠肺上皮细胞的抗氧化能力  

SIRT5 increasing the antioxidant capacity of COPD mouse lung epithelial cells by promoting IDH2 desuccinylation

作　　者：蒙婷 米叶斯尔·买买提艾力 徐丹[3] 李争[3] 李风森[1,3] MENG Ting;Miyesier Maimaiti Aili;XU Dan;LI Zheng;LI Feng-sen(College of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi Xinjiang 830000,China;Department of Internal Medicine,Eighth People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi Xinjiang 830000,China;National Clinical Research Base of Traditional Chinese Medicine,Fourth Affiliated Hospital of Xinjiang Medical University,Urumqi Xinjiang 830000,China)

机构地区：[1]新疆医科大学中医学院,新疆乌鲁木齐830011 [2]新疆维吾尔自治区第八人民医院内科,新疆乌鲁木齐830000 [3]新疆医科大学第四附属医院国家中医临床研究基地,新疆乌鲁木齐830000

出　　处：《局解手术学杂志》2023年第3期211-216,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：新疆维吾尔自治区自然科学基金(2021D01A143);2022年新疆维吾尔自治区青年岐黄学者项目。

摘　　要：目的探讨去乙酰化酶5(SIRT5)对慢性阻塞性肺病(COPD)中肺上皮细胞抗氧化能力的调节作用及机制。方法体外实验:肺上皮细胞MLE-12经H_(2)O_(2)、SIRT5过表达质粒(SIRT5-OE)或空载体(EV)处理后分为control组、H_(2)O_(2)组、SIRT5-OE组、EV组、H_(2)O_(2)+SIRT5-OE组、H_(2)O_(2)+EV组。在体实验:用烟草烟雾(CS)和脂多糖(LPS)诱导小鼠COPD模型,并分为正常组(小鼠不进行任何处理)、CS组(模型小鼠)、CS+saline组(生理盐水静脉注射到模型小鼠中)、CS+rhSIRT5组(将rhSIRT5静脉注射到模型小鼠中)、CS+rhSIRT5+SIRT5 inhibitor 1组(将rhSIRT5和SIRT5抑制剂共同注射到模型小鼠中)。按照说明书检测各组肺细胞/肺组织中丙二醛(MDA)、活性氧(ROS)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的水平;流式细胞术检测细胞凋亡水平。免疫沉淀法和蛋白免疫印迹法检测柠檬酸脱氢酶2(IDH2)的琥珀酰化水平和SIRT5蛋白表达。结果与EV组相比,SIRT5-OE组抑制IDH2的琥珀酰化水平降低,SIRT5蛋白表达增加,差异均有统计学意义(P<0.05)。与control组相比,H_(2)O_(2)组中IDH2的琥珀酰化、凋亡率、MDA、ROS的水平都增加/高(P<0.05)。与H_(2)O_(2)+EV组相比,H_(2)O_(2)+SIRT5-OE组的琥珀酰化和细胞凋亡率降低,且SOD和GSH-Px水平增加,差异均有统计学意义(P<0.05)。与正常组相比,CS组肺组织中IDH2的琥珀酰化、MDA、ROS的水平均增加/高(P<0.05)。与CS+saline组相比,CS+rhSIRT5组中IDH2的琥珀酰化减少,而SOD和GSH-Px水平均增加,差异均有统计学意义(P<0.05)。与CS+rhSIRT5组相比,CS+rhSIRT5+SIRT5 inhibitor 1组中MDA、ROS的水平均增加,SOD、GSH-Px水平明显降低,差异均有统计学意义(P<0.05)。结论SIRT5通过促进IDH2去琥珀酰化增加COPD小鼠肺上皮细胞的抗氧化能力。Objective To investigate the regulatory effect of sirtuin 5(SIRT5)on the antioxidant capacity of lung epithelial cells in chronic obstructive pulmonary disease(COPD)and its mechanism.Methods In vitro experiment:lung epithelial cells MLE-12 were treated with H_(2)O_(2),SIRT5 over-expression(SIRT5-OE)or empty vector(EV)and then divided into the control group,H_(2)O_(2)group,SIRT5-OE group,EV group,H_(2)O_(2)+SIRT5-OE group and H_(2)O_(2)+EV group.In vivo experiment:the mice were treated by cigarette smoke(CS)and lipopolysaccharide(LPS)for inducing the establishment of COPD model,and they were divided into the normal group(mice without any treatment),CS group(model mice),CS+saline group(injected saline intravenously after modeling),CS+rhSIRT5 group(injected rhSIRT5 intravenously after modeling)and CS+rhSIRT5+SIRT5 inhibitor 1 group(co-injected rhSIRT5 and SIRT5 inhibitor after modeling).The levels of malondialdehyde(MDA),reactive oxygen species(ROS),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in lung cells/tissues of each group were detected according to the instructions.The level of apoptosis was detected by flow cytometry.The succinylation level of isocitrate dehydrogenase 2(IDH2)and expression of SIRT5 protein were detected by immunoprecipitation and Western blot.Results Compared with the EV group,the inhibitation of the succinylation level of IDH2 in the SIRT5-OE group was decreased,and the expression of SIRT5 protein were increased,with statistically significant difference(P<0.05).Compared with the control group,the levels of IDH2 succinylation,apoptosis rate,MDA and ROS in the H_(2)O_(2)group were all increased(P<0.05).Compared with the H_(2)O_(2)+EV group,the succinylation and apoptosis rate of the H_(2)O_(2)+SIRT5-OE group were decreased,and the levels of SOD and GSH-Px were increased,with statistically significant differences(P<0.05).Compared with the normal group,the levels of IDH2 succinylation,MDA and ROS in the lung tissue in the CS group were all increased(P<0.05).Compared with the CS+sa

关 键 词：去琥珀酰化 去乙酰化酶5 慢性阻塞性肺病 小鼠 肺上皮细胞 柠檬酸脱氢酶2 

分 类 号：R563.3[医药卫生—呼吸系统]