利拉鲁肽抑制高糖诱导的小鼠胰岛细胞损伤  

Liraglutide Inhibits High Glucose-induced Injury of Mouse Islet Cells

作　　者：朱谋 白明健 王琪琦 程昊[2] 李林[3] 徐晶[3] 张春晶[3] ZHU Mou;BAI Ming-Jian;WANG Qi-Qi;CHENG Hao;LI Lin;XU Jing;ZHANG Chun-Jing(Qiqihar Medical University,Qiqihar 161006,Heilongjiang,China;Health Center,Qiqihar Medical University,Qiqihar 161006,Heilongjiang,China;Department of Biochemistry,Qiqihar Medical University,Qiqihar 161006,Heilongjiang,China)

机构地区：[1]齐齐哈尔医学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔医学院卫生所,黑龙江齐齐哈尔161006 [3]齐齐哈尔医学院生物化学教研室,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8期1062-1069,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：黑龙江省自然科学基金项目(No.LH2020H129);黑龙江省大学生创新创业训练计划项目(No.202111230004)资助。

摘　　要：利拉鲁肽(liraglutide,Lira)是胰高血糖素样肽-1的类似物,在糖尿病治疗中发挥重要作用,但利拉鲁肽通过改善胰岛β细胞的功能实现治疗糖尿病的具体机制尚未完全阐明。本研究采用高糖(33 mmol/L)诱导胰岛MIN6细胞48 h建立高糖损伤模型,CCK-8检测发现,与对照组相比,高糖组MIN6细胞活力下降(P<0.05),利拉鲁肽作用高糖组细胞活力升高(P<0.05);小鼠胰岛素和ATP含量检测发现,与对照组相比,高糖组胰岛素分泌降低(P<0.01),ATP含量减少(P<0.001),利拉鲁肽作用高糖组胰岛素释放量增加(P<0.05)和细胞内ATP含量增加(P<0.001);采用活体细胞线粒体膜通道孔(MPTP)荧光检测发现,与对照组相比,高糖组绿色荧光强度降低(P<0.001),利拉鲁肽作用高糖组绿色荧光强度增加(P<0.001);DCFH-DA探针联合流式细胞仪检测细胞活性氧簇(ROS)含量发现,与对照组相比,高糖组ROS水平升高(P<0.001),利拉鲁肽作用高糖组ROS水平降低(P<0.01);细胞内丙二醛(MDA)含量、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)以及细胞上清中乳酸脱氢酶(LDH)活性测定发现,与对照组相比,高糖组MDA和LDH水平升高(P<0.05),SOD和CAT水平降低(P<0.01),利拉鲁肽作用高糖组细胞内MDA含量和LDH活性降低(P<0.05),SOD和CAT活性增加(P<0.05);Western印迹检测解偶联蛋白2(uncoupling protein 2,UCP2)的表达发现,与对照组相比,高糖组UCP2表达上调(P<0.01),利拉鲁肽作用高糖组UCP2表达降低(P<0.05)。结果表明,利拉鲁肽对高糖诱导MIN6细胞的线粒体损伤、氧化应激以及胰岛素分泌具有重要作用,其作用机制可能与下调UCP2的表达相关,为利拉鲁肽更好地应用于临床提供了理论依据。Liraglutide is an analog of glucagon-like peptide-1 and plays an important role in the treatment of diabetes.But the specific mechanism of liraglutide in improving the function of pancreaticβcells is still unclear.Therefore,high glucose(33 mmol/L)was used to induce islet MIN6 cells for 48 hours to establish a high glucose injury model in this study.The CCK-8 assay was used to detect cell viability,and the results showed that the viability of MIN6 cells in the high glucose group decreased(P<0.05)compared with the control group,and the cell viability increased after liraglutide treatment(P<0.05).Using the mouse insulin detection kit and ATP content detection kit,we found that both the insulin release(P<0.01)and ATP content decreased(P<0.001)in the high glucose group compared with the control group,and after liraglutide treatment both insulin release(P<0.05)and ATP content(P<0.001)increased.We used the mitochondrial membrane channel pore(MPTP)fluorescence detection kit in MIN6 cells and found that the green fluorescence intensity of the high glucose group was significantly decreased(P<0.001)compared with the control group,and after liraglutide treatment the green fluorescence intensity was significantly increased(P<0.001).The DCFH-DA probe combined with flow cytometry was used to detect the level of reactive oxygen species(ROS).We found that compared with the control group,the ROS level in the high glucose group was significantly increased(P<0.001),and decreased by liraglutide treatment(P<0.01).Intracellular malondialdehyde(MDA)contents,superoxide dismutase(SOD)and catalase(CAT),and lactate dehydrogenase(LDH)activities in the cell supernatant were measured,and the levels of MDA and LDH were significantly increased(P<0.05),and the levels of SOD and CAT were significantly decreased(P<0.01)in the high glucose group compared with the control group.After liraglutide treatment,the levels of MDA and LDH were decreased(P<0.05),and the levels of SOD and CAT were increased(P<0.05).The results of Western blotting showed that

关 键 词：利拉鲁肽 小鼠胰岛细胞 高糖 线粒体损伤 

分 类 号：Q51[生物学—生物化学]