悬钩子苷对2型糖尿病脂毒性Min6细胞凋亡模型的影响研究  

Effect of Rubusoside on palmitic acid-induced apoptosis in mouse pancreatic isletβ-cells

作　　者：徐文悦 赵娜[2] 姜新[3] 宋苏丽 王燕[2] 董明鑫 张剑旭 孙成彪 刘文森 许娜[3] 王龙涛[1] XU Wen-yue;ZHAO Na;JIANG Xin;SONG Su-li;WANG Yan;DONG Ming-xin;ZHANG Jian-xu;SUN Cheng-biao;LIU Wen-sen;XU Na;WANG Long-tao(College of Animal Medicine,Jilin A gricultural University,Changchun 130118,China;Changchun Institute of Veterinary Medicine,Chinese Academy of Agricultural Sciences,Changchun 130000,China;Jilin Medical College,Jilin 132000,China;Changchun University of Science and Technology,Changchun 130022,China)

机构地区：[1]吉林农业大学动物医学院,吉林长春130118 [2]中国农业科学院长春兽医研究所,吉林长春130000 [3]吉林医药学院,吉林吉林132000 [4]长春理工大学,吉林长春130022

出　　处：《中国兽医科学》2023年第1期114-120,共7页Chinese Veterinary Science

基　　金：吉林省教育厅科研课题(JJKH20210503KJ)。

摘　　要：为探讨悬钩子苷(Rub)对棕榈酸(PA)诱导的小鼠胰岛β细胞(Min6)凋亡的影响,首先利用PA诱导的Min6细胞建立2型糖尿病(T2DM)脂毒性细胞模型,分别给予不同浓度Rub对细胞模型进行干预,分为对照组、模型组、PA+Rub组(PA+12.5μmol/L Rub组、PA+50μmol/L Rub组、PA+100μmol/L Rub组和PA+200μmol/L Rub组)。通过Annexin V-FITC和PI双染流式细胞术检测Rub对PA诱导的Min6细胞凋亡率的影响;采用RT-PCR检测各组细胞凋亡相关基因Caspase 3、Bcl-2和Bax m RNA的表达;通过免疫荧光技术检测Min6细胞内Caspase 9的表达;Western-blot检测Caspase 9、Caspase 3、Cleaved-Caspase 9、Cleaved-Caspase 3、Bcl-2和Bax蛋白表达;ELISA法检测胰岛素分泌水平。结果显示,与对照组相比,模型组胰岛素水平及细胞中Bcl-2/Bax m RNA及蛋白表达量均显著降低(P<0.01),细胞凋亡率及细胞中Caspase3 m RNA表达量和Cleaved-Caspase 9/Caspase 9、Cleaved-Caspase 3/Caspase 3蛋白表达量均显著升高(P<0.01)。与模型组相比,PA+Rub组细胞上述指标均发生明显改善,且呈剂量依赖性(P<0.05或P<0.01)。证实Rub具有抑制胰岛β细胞凋亡的作用,其分子机制可能与细胞凋亡的线粒体途径有关。In order to investigate the effect of Rubusoside(Rub)on palmitic acid(PA)-induced apoptosis of mouse pancreaticβcells(Min6).Firstly,PA-induced Min6 cells were used to establish type2 diabetes mellitus(Type 2 diabetes mellitus,T2DM)lipotoxicity cell model,different concentrations of Rub were given to intervene in the cell model,and they were divided into Control group,Model group,PA+Rub group(PA+12.5μmol/L Rub group,PA+50μmol/L Rub group,PA+100μmol/L Rub group,and PA+200μmol/L Rub group).The effect of Rub on the rate of apoptosis of PA-induced Min6 cells was detected by Annexin V-FITC and PI double-stained flow cytometry.RT-PCR was used to detect the m RNA expression of apoptosis-related genes Caspase 3,Bcl-2,and Bax in each group.The Caspase 9 expression in Min6 cells was assessed using immunofluorescence,the protein level of Caspase 9,Caspase 3,Cleaved-Caspase 9,Cleaved-Caspase 3,Bcl-2 and Bax were tested by Western-blot.The level of insulin secretion was detected by ELISA.The result showed that compared with the Control group,the level of insulin and the m RNA and protein expression of Bcl-2/Bax in the Model group were significantly decreased(P<0.01),as well as the cell apoptosis rate,m RNA expression of Caspase 3 and the protein expression of CleavedCaspase 9/Caspase 9,Cleaved-Caspase 3/Caspase 3,were significantly increased(P<0.01).Compared with the Model group,all of the above indexes of cells in the PA+Rub group were significantly improved in a dose-dependent manner(P<0.05 or P<0.01).In conclusion,Rub could significantly inhibit the apoptosis of Min6 cells induced by PA,and the molecular mechanism may be related to the mitochondrial pathway of apoptosis.

关 键 词：悬钩子苷 2型糖尿病 细胞凋亡 棕榈酸 脂毒性 

分 类 号：S853.74[农业科学—临床兽医学]