GLP-1通过细胞色素P450表氧化酶途径抑制氧化应激

doi:10.6040/j.issn.1671-7554.0.2023.0450

摘要/Abstract

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 H₂O₂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、H₂O₂组、H₂O₂+7-36a组、H₂O₂+9-36a组、H₂O₂+7-36a+Danazol(CYP2J2蛋白抑制剂)组、H₂O₂+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 600 μmol/L的H₂O₂处理HUVECs 3 h建立氧化应激模型,与对照组比较,H₂O₂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H₂O₂组比较,H₂O₂+7-36a组和H₂O₂+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H₂O₂+7-36a组相比,H₂O₂+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H₂O₂+9-36a组相比,H₂O₂+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H₂O₂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。

关键词: 人脐静脉内皮细胞, 氧化应激, GLP-1, 细胞色素P450

Abstract: 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 H₂O₂ to construct the oxidative stress models. The cells were divided into six groups: control group, H₂O₂ group, H₂O₂+7-36a group, H₂O₂+9-36a group, H₂O₂+7-36a+Danazol group and H₂O₂+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 P450 2J2(CYP2J2)were detected with Western blotting. Results Oxidative stress models were constructed after 600 μmol/L H₂O₂ treatment for 3 hours. Compared with the control group, the H₂O₂ 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 H₂O₂ group, the H₂O₂+7-36a group and H₂O₂+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 H₂O₂+7-36a group, there was no statistical difference in cell survival rate in the H₂O₂+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 H₂O₂+9-36a group, there was no statistical difference in cell survival rate in the H₂O₂+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 H₂O₂-induced survival decline in HUVECs and reduce the high level of oxidative stress, possibly through the CYP450 epoxygenase pathway.

Key words: Human umbilical vein endothelial cells, Oxidative stress, GLP-1, Cytochrome P450

中图分类号:

R589

扈艳雯,赵蕙琛,马小莉,刘元涛,张玉超. GLP-1通过细胞色素P450表氧化酶途径抑制氧化应激[J]. 山东大学学报 (医学版), 2023, 61(8): 10-16.

HU Yanwen, ZHAO Huichen, MA Xiaoli, LIU Yuantao, ZHANG Yuchao. GLP-1 inhibits oxidative stress damage through cytochrome P450 surface oxidase pathway[J]. Journal of Shandong University (Health Sciences), 2023, 61(8): 10-16.

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