利拉鲁肽对Kir6.2基因E23K位点突变β细胞胰岛素分泌功能的影响  

The influence of liraglutide on insulin secretion function of E23K mutation of Kir6.2 gene inβcells

作　　者：庄兰艮 金国玺[1] 杨青青[1] 胡小磊[1] 裴晓艳[1] ZHUANG Langen;JIN Guoxi;YANG Qingqing;HU Xiaolei;PEI Xiaoyan(Department of Endocrinology,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内分泌科,安徽蚌埠233004

出　　处：《中华全科医学》2023年第3期385-388,共4页Chinese Journal of General Practice

基　　金：蚌埠医学院转化医学重点专项项目(BYTM2019032);安徽省自然科学基金青年项目(1908085QH319)。

摘　　要：目的Kir6.2编码基因多态与糖尿病、胰岛素抵抗相关,本研究旨在探讨高糖环境下,利拉鲁肽对Kir6.2基因E23K位点突变的胰岛β细胞胰岛素分泌功能的影响及机制。方法以NIT-1细胞为研究对象,构建Kir6.2基因过表达野生型和E23K突变型NIT-1细胞,并用利拉鲁肽在高糖环境下进行24 h干预。采用流式细胞仪检测NIT-1细胞凋亡率、细胞膜电位及钙离子浓度,Western blotting和免疫荧光检测细胞内胰岛素含量,ELISA检测培养液中胰岛素含量,并以此间接研究胰岛素的分泌情况。结果NIT-1细胞最适高糖培养浓度为60 mmol/L,在此浓度下胰岛素合成量最高,利拉鲁肽体外干预浓度选择10 mmol/L。过表达野生型Kir6.2基因NIT-1细胞的胰岛素分泌高于Kir6.2基因E23K突变型NIT-1细胞。过表达野生型Kir6.2基因NIT-1细胞在高糖环境中细胞膜电位进一步降低,细胞内Ca^(2+)含量增加,而Kir6.2基因E23K突变型NIT-1细胞与单纯高糖培养相比无显著改变。利拉鲁肽在高糖环境中可有效减少野生型和突变型NIT-1细胞的凋亡,降低膜电位,增加细胞内Ca^(2+)含量,促进胰岛素分泌。另外利拉鲁肽对维持突变型NIT-1细胞存活作用更显著。结论在高糖环境中,利拉鲁肽能改善Kir6.2基因E23K位点多态型胰岛β细胞的胰岛素分泌功能,该研究为利拉鲁肽的临床应用提供基础研究证据。Objective Polymorphism of Kir6.2 is associated with diabetes and insulin resistance.This study aims to explore whether liraglutide can improve the insulin secretion function and mechanism of the E23K mutation of Kir6.2 gene ofβcell of islet under a high glucose condition.Methods Taking NIT-1 cells as the object,Kir6.2 gene-overexpressed wild-type and E23K mutant NIT-1 cells were constructed,and liraglutide was adopted to intervene in a high-glucose environment for 24 hours.Flow cytometry was used to detect the NIT-1 cell apoptosis rate,the cell membrane potential and the calcium ion concentration.Western blotting and immunofluorescence assay were conducted to detect insulin contents in cells.ELISA was utilized to detect insulin contents in nutrient fluid,based on which insulin secretion was indirectly studied.Results NIT-1 cells had an optimal high glucose culture concentration of 60 mmol/L,with the highest amount of insulin synthesis,whereas 10 mmol/L was selected for liraglutide in vitro.The insulin secretion of Kir6.2 gene-overexpressed wild-type NIT-1 cells was higher than that of Kir6.2 gene E23K mutant NIT-1 cells.The Kir6.2 gene-overexpressed wild-type NIT-1 cells presented less membrane potential and higher Ca^(2+) concentration in cells in a high-glucose environment,while Kir6.2 gene E23 mutant NIT-1 cells made no significant changes compared with those cultured under high glucose concentration.Liraglutide could effectively reduce apoptosis of wild-type and mutant NIT-1 cells,decrease membrane potential,increase Ca^(2+) concentration in the cells,and promote insulin secretion in a high-glucose environment.In addition,liraglutide was of greater significance to the survival of mutant NIT-1 cells.Conclusion In a high-glucose environment,liraglutide can improve the insulin secretion function of polymorphic of E23K site Kir6.2 gene isletβcell.This study provides basic evidence for the clinical application of liraglutide.

关 键 词：利拉鲁肽 KIR6.2 E23K突变 胰岛Β细胞 胰岛素 

分 类 号：R587[医药卫生—内分泌]