白藜芦醇对Gc-1spg细胞氧化应激损伤的作用研究  

Effect of resveratrol on oxidative stress injury of Gc-1 spg cells

作　　者：石拴霞 阎一鑫 宋诚 王纪田 何毅刚 王玲 Shi Shuan-xia;Yan Yi-xin;Song Cheng;Wang Ji-tian;He Yi-gang;Wang Ling(No.1 School of Clinical Medicine,Gansu University of traditional Chinese Medicine,Lanzhou,Gansu 730000,China;Reproductive Medicine Centre,The 940th Hospital of Joint Logistic Support Force of Chinese People's Liberation Army,Lanzhou,Gansu 730050,China;Key Laboratory of Stem Cells and Gene Drugs of Gansu Province,The 940th Hospital of Joint Logistic Support Force of Chinese People's Liberation Army,Lanzhou,Gansu 730050,China)

机构地区：[1]甘肃中医药大学第一临床医学院,甘肃兰州730000 [2]中国人民解放军联勤保障部队第九四〇医院生殖医学中心,甘肃兰州730050 [3]中国人民解放军联勤保障部队第九四〇医院甘肃省干细胞与基因药物重点实验室,甘肃兰州730050

出　　处：《中国现代医学杂志》2023年第19期46-54,共9页China Journal of Modern Medicine

基　　金：甘肃省青年科技基金计划(No:21JR1RA188);军队后勤科研计生专项课题(No:20JSZ09)。

摘　　要：目的观察白藜芦醇(RES)对过氧化氢H_(2)O_(2)诱导的小鼠精原细胞(Gc-1 spg)氧化应激损伤的作用。方法H_(2)O_(2)复制Gc-1 spg细胞氧化应激损伤模型,设置空白组、H_(2)O_(2)组(800µmol/L)、H_(2)O_(2)+5µmol/L RES组、H_(2)O_(2)+10µmol/L RES组和H_(2)O_(2)+15µmol/L RES组。检测超氧化物歧化酶(SOD)、丙二醛(MDA)、乳酸脱氢酶(LDH)和谷胱甘肽过氧化物酶(GSH-Px)水平,CCK-8法检测细胞活力,JC-1试剂盒检测线粒体膜电位,Mito Tracker®®Green FM试剂盒检测活细胞线粒体水平,Hoechst 33342染色检测细胞核凋亡率,Western blotting检测凋亡蛋白Bax、Bcl-2和Caspase-3表达。结果浓度为800µmol/L H_(2)O_(2)处理Gc-1 spg细胞6 h时达到实验要求(P<0.05)。与空白组比较,各H_(2)O_(2)组细胞活力降低(P<0.05),与H_(2)O_(2)组比较,H_(2)O_(2)+5µmol/L RES组、H_(2)O_(2)+10µmol/L RES组、H_(2)O_(2)+15µmol/L RES组细胞活力升高(P<0.05)。与空白组比较,各H_(2)O_(2)组的GSH-Px和SOD水平降低(P<0.05),LDH和MDA水平升高(P<0.05);与H_(2)O_(2)组比较,H_(2)O_(2)+5µmol/L RES组、H_(2)O_(2)+10µmol/L RES组、H_(2)O_(2)+15µmol/L RES组GSH-PX和SOD水平降低(P<0.05),LDH和MDA水平升高(P<0.05)。与空白组比较,H_(2)O_(2)组细胞的红/绿荧光比值降低,提示线粒体膜电位降低(P<0.05);与H_(2)O_(2)组比较,H_(2)O_(2)+5µmol/L RES组、H_(2)O_(2)+10µmol/L RES组、H_(2)O_(2)+15µmol/L RES组JC-1红/绿荧光比值升高(P<0.05)。与空白组比较,H_(2)O_(2)组细胞线粒体数明显减少(P<0.05);与H_(2)O_(2)组比较,H_(2)O_(2)+5µmol/L RES组、H_(2)O_(2)+10µmol/L RES组、H_(2)O_(2)+15µmol/L RES组细胞荧光强度升高(P<0.05)。与空白组比较,H_(2)O_(2)组大量细胞核皱缩、碎裂,染色程度明显增强,细胞凋亡严重;与H_(2)O_(2)组比较,H_(2)O_(2)+5µmol/L RES组、H_(2)O_(2)+10µmol/L RES组、H_(2)O_(2)+15µmol/L RES组细胞凋亡率降低(P<0.05)。与空白组比较,H_(2)O_(2)组凋亡蛋白Bax和Caspase-3相对表达量增加(P<0.05),BcObjective To observe the effect of resveratrol(RES)on oxidative stress injury of mouse spermatogonia(Gc-1 spg)induced by hydrogen peroxide(H_(2)O_(2)).Methods Set the blank control group,H_(2)O_(2) group and H_(2)O_(2)+low concentration RES group(5μmol/L),H_(2)O_(2)+medium concentration RES group(10μmol/L)and H_(2)O_(2)+high concentration RES group(15μmol/L),the RES group was treated with RES of different concentrations for 24 h,and the rest groups were treated with H_(2)O_(2) for 6 h except the control group.The levels of oxidative stress indicators SOD,MDA,GSH-Px and LDH were detected,and the cell viability was detected by CCK-8,detection of mitochondrial membrane potential by JC-1,Mito Tracker®Green FM was used to detect the level of mitochondria in living cells,Hoechst 33342 staining for detection of nuclear apoptosis rate and Western blotting was used to detect the expression level of apoptotic proteins Bax,Bcl-2 and Caspase-3.Results Concentration is 800μmol/L when Gc-1 spg cells were treated with H_(2)O_(2) for 6 h,the experimental requirements were met(P<0.05).Compared with the control group,H_(2)O_(2) treatment significantly decreased cell viability(P<0.05),inhibited SOD and GSH-Px activities,increased MDA and LDH levels(P<0.05),significantly decreased mitochondrial membrane potential(P<0.05),damaged mitochondria(P<0.05)and destroy nucleus(P<0.05),increased the expression levels of Bax and Caspase-3,and decreased the expression level of Bcl-2(P<0.05).Below 20μmol/L RES had no significant effect on cell viability(P>0.05),partially restored cell viability(P<0.05),increased SOD and GSH-Px activities,decreased MDA and LDH levels(P<0.05),increased mitochondrial membrane potential(P<0.05),reduced mitochondrial damage(P<0.05)and apoptosis rate,the expression of Bax and Caspase-3 was decreased and the expression of Bcl-2 was increased in a concentration dependent manner(P<0.05).Conclusion RES can protect Gc-1 spg cells from oxidative stress injury induced by H_(2)O_(2),which is related to the concentration

关 键 词：白藜芦醇 过氧化氢 Gc-1 spg细胞 氧化应激 线粒体损伤 凋亡 

分 类 号：R698.2[医药卫生—泌尿科学]