活性维生素D对胰岛β细胞自噬及相关蛋白表达的影响  

Effects of active Vitamin D on autophagy and related protein expression in pancreaticβcells

作　　者：师佩兰[1] 郭道华[1] 余美玲[1] 许健[1] 宋乐乐[1] 桑冉[1] Shi Peilan;Guo Daohua;Yu Meiling;Xu Jian;Song Lele;Sang Ran(Department of pharmacy,the first affiliated hospital of Bengbu Medical College,Bengbu,Anhui,233004,China)

机构地区：[1]蚌埠医学院第一附属医院药剂科,安徽蚌埠233004

出　　处：《齐齐哈尔医学院学报》2022年第17期1614-1617,共4页Journal of Qiqihar Medical University

基　　金：蚌埠医学院自然科学重点项目(2020byzd079)。

摘　　要：目的探讨活性维生素D(VitD)对高糖环境下胰岛β细胞自噬及相关蛋白表达的影响。方法体外培养Min6小鼠胰岛β细胞,将细胞分为5组:对照组(Con,5.6 mmol/L葡萄糖)、高糖组(HG,30 mmol/L葡萄糖)、高糖+1 nmol/L活性VitD组(HG+1VitD)、高糖+10 nmol/L活性VitD组(HG+10VitD)、高糖+100 nmol/L活性VitD组(HG+100VitID)。采用MTT法检测细胞存活率,免疫荧光(IF)法观察自噬相关蛋白LC3-Ⅱ含量的变化,Western-Blot法检测AMPK、LC3蛋白的表达。结果与对照组比较,HG组及HG+VitD组细胞存活率降低(P<0.05),HG+VitD组AMPK、LC3蛋白表达升高(P<0.05);与HG组相比,HG+VitD组细胞存活率、自噬相关蛋白LC3-Ⅱ含量及AMPK、LC3蛋白表达呈浓度依赖性升高(P<0.05)。结论活性VitD可改善高糖环境下胰岛β细胞的生长,其机制可能与调控AMPK通路诱导自噬相关。Objective To investigate effect of active Vitamin D on autophagy and related protein expression of pancreaticβcells under high glucose concentration.Methods The cultured Min6 mouse isletβcells were divided into 5 groups:the control group(Con,5.6 mmol/L glucose),high glucose group(HG,30 mmol/L glucose),high glucose+1 nmol/L active VitD group(HG+1 VitD),high glucose+10 nmol/L active VitD group(HG+10 VitD),high glucose+100 nmol/L active VitD group(HG+100 VitD).The cell survival rate was detected by MTT assay,the content of autophagyassociated proteinLC3-Ⅱwas observed by immunofluorescence(IF),the expression of AMPK and LC3 protein was detected by Western Blot.Results Compared with control group,the cell survival rate in HG and HG+VitD groups decreased(P<0.05),the expression of AMPK and LC3 protein in HG+VitD groups increased(P<0.05).Compared with HG group,cell survival rate,LC3-Ⅱ content and expression of AMPK,LC3 protein in HG+VitD groups increased in a dose-dependent manner(P<0.05).Conclusions Active VitD could improve the growth of islet β cells cultured in high glucose concentration,and the mechanism may be related to the regulation of AMPK pathway inducing autophagy.

关 键 词：活性维生素D 胰岛Β细胞 自噬 LC3 AMPK 

分 类 号：R335[医药卫生—人体生理学]