GLP-1通过细胞色素P450表氧化酶途径抑制氧化应激  

GLP-1 inhibits oxidative stress damage through cytochrome P450 surface oxidase pathway

作　　者：扈艳雯 赵蕙琛 马小莉[2] 刘元涛 张玉超[2] HU Yanwen;ZHAO Huichen;MA Xiaoli;LIU Yuantao;ZHANG Yuchao(Department of Nutrition,Heze Municipal Hospital,Heze 274000,Shandong,China;Department of Endocrinology,Qingdao Municipal Hospital,Qingdao 266011,Shandong,China;Department of Endocrinology,Qilu Hospital(Qingdao),Cheeloo College of Medicine,Shandong University,Qingdao 266035,Shandong,China)

机构地区：[1]菏泽市立医院营养科,山东菏泽274000 [2]青岛市市立医院内分泌科,山东青岛266011 [3]山东大学齐鲁医院(青岛)内分泌科,山东青岛266035

出　　处：《山东大学学报（医学版）》2023年第8期10-16,共7页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(ZR202111120136)。

摘　　要：目的研究GLP-1在体外对氧化应激的保护作用,并从细胞色素P450(CYP450)表氧化酶途径探讨可能的机制。方法H_(2)O_(2)处理人脐静脉内皮细胞(HUVECs)构建氧化应激模型;实验分为对照组、H_(2)O_(2)组、H_(2)O_(2)+7-36a组、H_(2)O_(2)+9-36a组、H_(2)O_(2)+7-36a+Danazol(CYP2J2蛋白抑制剂)组、H_(2)O_(2)+9-36a+Danazol组。CCK8方法检测细胞存活率;荧光探针标记法测定细胞内ROS水平;Western blotting方法检测葡萄糖调节蛋白78(GRP78)及细胞色素P450表氧化酶2J2(CYP2J2)的表达变化。结果600μmol/L的H_(2)O_(2)处理HUVECs 3 h建立氧化应激模型,与对照组比较,H_(2)O_(2)组细胞存活率下降(P<0.001),ROS水平升高(P<0.001),GRP78蛋白表达量增加(P<0.05),CYP2J2蛋白表达量下降(P<0.05)。与H_(2)O_(2)组比较,H_(2)O_(2)+7-36a组和H_(2)O_(2)+9-36a组细胞存活率均上升(P<0.05;P<0.01),ROS水平均下降(P<0.001),GRP78表达量均减少(P<0.01),CYP2J2表达量均增加(P<0.05)。加入Danazol后,与H_(2)O_(2)+7-36a组相比,H_(2)O_(2)+7-36a+Danazol组的细胞存活率差异无统计学意义,ROS水平升高(P<0.001),GRP78蛋白表达增加(P<0.001),CYP2J2表达降低(P<0.05);与H_(2)O_(2)+9-36a组相比,H_(2)O_(2)+9-36a+Danazol组的细胞存活率差异无统计学意义,ROS水平升高(P<0.001),GRP78蛋白表达增加(P<0.01),CYP2J2表达降低(P<0.01)。结论GLP-1(7-36a)及其代谢产物GLP-1(9-36a)改善H_(2)O_(2)诱导的HUVECs细胞存活率下降与高氧化应激水平,其可能通过CYP450表氧化酶途径发挥抑制氧化应激的作用。Objective To explore the effects of GLP-1 against oxidative stress in vitro and the possible mechanism from cytochrome P450(CYP450)epoxygenase pathway.Methods Human umbilical vein endothelial cells(HUVECs)were treated with H_(2)O_(2)to construct the oxidative stress models.The cells were divided into six groups:control group,H_(2)O_(2)group,H_(2)O_(2)+7-36a group,H_(2)O_(2)+9-36a group,H_(2)O_(2)+7-36a+Danazol group and H_(2)O_(2)+9-36a+Danazol group.Cell viability was evaluated with CCK8 assay.Intracellular ROS level was determined with fluorescent probe labeling.The expressions of glucose regulatory protein 78(GRP78)and cytochrome P4502J2(CYP2J2)were detected with Western blotting.Results Oxidative stress models were constructed after 600μmol/L H_(2)O_(2)treatment for 3 hours.Compared with the control group,the H_(2)O_(2)group had decreased cell survival rate(P<0.001),increased ROS level(P<0.001)and protein expression of GRP78(P<0.05),but decreased CYP2J2 protein expression(P<0.05).Compared with the H_(2)O_(2)group,the H_(2)O_(2)+7-36a group and H_(2)O_(2)+9-36a group had increased survival rate(P<0.05,P<0.01),decreased ROS level(both P<0.001)and GRP78 protein expression(both P<0.01),and increased CYP2J2 protein expression(both P<0.05).Compared with the H_(2)O_(2)+7-36a group,there was no statistical difference in cell survival rate in the H_(2)O_(2)+7-36a+Danazol group,while the ROS level increased(P<0.001),GRP protein expression incresed(P<0.001),and CYP2J2 protein expression decreased(P<0.05).Compared with the H_(2)O_(2)+9-36a group,there was no statistical difference in cell survival rate in the H_(2)O_(2)+9-36a+Danazol group,while the ROS level increased(P<0.001),GRP protein expression incresed(P<0.01),and CYP2J2 protein expression decreased(P<0.01).Conclusion GLP-1(7-36a)and its metabolite GLP-1(9-36a)can improve H_(2)O_(2)-induced survival decline in HUVECs and reduce the high level of oxidative stress,possibly through the CYP450 epoxygenase pathway.

关 键 词：人脐静脉内皮细胞 氧化应激 GLP-1 细胞色素P450 

分 类 号：R589[医药卫生—内分泌]