京尼平保护高糖诱导损伤小鼠胰岛MIN6细胞的机制  

Genipin Protects High Glucose-injured MIN6 Cells

作　　者：徐晶[1] 赵正林 师岩[1] 高涵[1] 刘哲丞[1] 武爽 朱谋 张春晶[1] XU Jing;ZHAO Zheng-Lin;SHI Yan;GAO Han;LIU Zhe-Cheng;WU Shuang;ZHU Mou;ZHANG Chun-Jing(Department of Biochemistry and Molecular Biology,Qiqihar Medical University,Qiqihar 161006,Heilongjiang,China)

机构地区：[1]齐齐哈尔医学院生化教研室,黑龙江齐齐哈尔161006

出　　处：《中国生物化学与分子生物学报》2022年第1期83-90,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：2018年黑龙江省教育厅基本科研业务费基础研究项目(No.2018-KYYWF-0083);黑龙江省自然科学基金项目(No.LH2020H129)。

摘　　要：京尼平(genipin,Gen)是一种重要的抗氧化物,在细胞内抵抗氧化应激损伤过程中发挥重要的作用。为了探讨京尼平对高糖诱导损伤的小鼠胰岛MIN6细胞的影响,采用CCK-8法检测细胞存活率。高糖损伤组细胞活力下降(P<0.05),京尼平作用高糖损伤的细胞后,细胞活力增加(P<0.05);小鼠胰岛素(insulin)检测试剂盒和ATP含量检测试剂盒检测发现,高糖损伤组胰岛素释放量减少(P<0.001),ATP含量减少(P<0.001),给予京尼平后,胰岛素释放量增加(P<0.01),ATP含量增加(P<0.01);荧光探针DCFH-DA检测细胞内活性氧簇(ROS)水平发现,高糖损伤组ROS含量明显增加(P<0.01),给予京尼平作用后,ROS含量减少(P<0.05);谷胱甘肽(GSH)/氧化型谷胱甘肽(GSSG)、细胞内丙二醛(MDA),超氧化物歧化酶(SOD)和过氧化氢酶(CAT)以及细胞上清液中乳酸脱氢酶(LDH)活性的测定发现,高糖损伤组细胞GSH/GSSH比值、SOD和CAT水平下降(P<0.05),细胞内MDA和LDH水平显著升高(P<0.001),给予京尼平后,GSH/GSSH比值、SOD和CAT水平增加(P<0.01),MDA和LDH水平显著降低(P<0.01);线粒体膜电位(JC-1)检测发现,高糖损伤组细胞中MMP水平下降(P<0.01),京尼平作用高糖损伤的细胞后,MMP水平增加(P<0.05);Western印迹检测解偶联蛋白2(uncoupling protein 2,UCP2)、抗氧化蛋白谷胱甘肽还原酶(glutathione reductase,GR)和谷氧还蛋白1(glutaredoxin 1,Grx1)含量结果显示,高糖损伤组UCP2含量增加(P<0.01)和抗氧化蛋白含量减少(P<0.01),给予京尼平后,UCP2表达下降(P<0.05)、抗氧化蛋白质的表达增加(P<0.05)。结果提示,京尼平对高糖损伤的小鼠胰岛细胞具有抗氧化性的保护作用,以及维持胰岛细胞功能促进胰岛素分泌的作用。本文为京尼平对高糖损伤的小鼠胰岛MIN6细胞的抗氧化作用提供实验数据,也为后续探讨京尼平在糖尿病的治疗和预防中提供新的思路。Genipin(Gen)is an important antioxidant that plays a crucial role in the process of intracellular resistance to oxidative stress.In order to study the effect of genipin on MIN6 cells injured by high glucose,the CCK-8 method was used to detect the cell survival rate.The cell viability of the high-glucose injury group decreased(P<0.05).But after genipin acted on the cells injured by high glucose,the cell viability increased(P<0.05).The mouse insulin detection kit and ATP content detection kit found that the insulin release in the high glucose injury group decreased(P<0.001)and the ATP content decreased(P<0.001).After genipin was given,the insulin release increased(P<0.01),ATP content increased(P<0.01).The fluorescent probe DCFH-DA detected the intracellular reactive oxygen species(ROS)level and found that the ROS content in the high glucose-injury group was significantly increased(P<0.01).After genipin was administered,ROS content decreased(P<0.05).Glutathione(GSH)/oxidized glutathione(GSSG),intracellular malondialdehyde(MDA),superoxide dismutase(SOD)and catalase(CAT)and lactate dehydrogenase(LDH)activity in the cell supernatant revealed that the GSH/GSSH ratio,SOD and CAT levels in the high glucose injury group decreased(P<0.05),and the intracellular MDA and LDH levels were significant increased(P<0.001).After administration of genipin,the GSH/GSSH ratio,SOD and CAT levels increased(P<0.01),MDA and LDH levels were significantly reduced(P<0.01).Mitochondrial membrane potential(MMP)levels decreased in the high glucose injury group(P<0.01).After genipin acted on the cells injured by high glucose,the MMP level increased(P<0.05).Western blot detected uncoupling protein 2(UCP2),antioxidative proteins glutathione reductase(GR)and glutaredoxin 1(Grx1)contents.The results showed that UCP2 contents in the high glucose injury group increased(P<0.01)and the content of oxidized protein decreased(P<0.01).After genipin was administered,the expression of UCP2 decreased(P<0.05),and the expression of antioxidative protein increased

关 键 词：京尼平 高糖 氧化应激 抗氧化 

分 类 号：Q51[生物学—生物化学]