文献类型：期刊文献

中文题名：黄连化浊胶囊对分泌及Ras相关GTP酶1A敲除诱导的外被蛋白Ⅱ功能障碍胰岛β细胞内质网应激的干预效果

英文题名：Intervention effects of Huanglian Huazhuo capsule on endoplasmic reticulum stress of islet β cells with coat protein Ⅱ dysfunction induced by secretion associated Ras related GTPase 1A knockdown

作者：周春楠[1];王苑铭[1]

第一作者：周春楠

机构：[1]甘肃中医药大学附属医院内分泌科,兰州730200

第一机构：甘肃中医药大学第二附属医院

年份：2023

卷号：44

期号：9

起止页码：1073

中文期刊名：海军军医大学学报

外文期刊名：Academic Journal of Naval Medical University

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：兰州市科技发展计划项目(2022-ZD-89).

语种：中文

中文关键词：胰岛β细胞;黄连化浊胶囊;内质网应激;分泌及Ras相关GTP酶1A

外文关键词：isletβcell;Huanglian Huazhuo capsule;endoplasmic reticulum stress;secretion associated Ras related GTPase 1A

摘要：目的考察黄连化浊胶囊对分泌及Ras相关GTP酶1A(sar-1A)敲除诱导的外被蛋白Ⅱ(COP-Ⅱ)功能障碍胰岛β细胞内质网应激的干预效果。方法将小鼠胰岛β细胞Min6分为空白组(Min6细胞无任何干预)、阴性对照(NC)组(Min6细胞转染sar-1A-NC-siRNA)及sar-1A siRNA组(Min6细胞转染sar-1A siRNA)。收集sar-1A siRNA转染的Min6细胞,分为空白血清组(用含10%大鼠空白血清的培养基培养),5%、10%、20%黄连化浊胶囊组(分别用含5%、10%、20%黄连化浊胶囊含药血清的培养基培养),以及罗格列酮组(用含10%罗格列酮含药血清的培养基培养)。采用qPCR检测Min6细胞中sar-1A miRNA的表达、免疫荧光法检测sec31蛋白的表达,蛋白质印迹法检测胰岛素原、磷酸化真核翻译起始因子2α(p-eIF2α)、C/EBP同源蛋白(CHOP)、X盒结合蛋白1(XBP1)、肌醇需求酶1(IRE1)、X盒(X-box)蛋白的表达。结果空白组与NC组Min6细胞中sar-1A mRNA、sec31蛋白及胰岛素原、p-eIF2α、CHOP、XBP1、IRE1、X-box蛋白表达差异均无统计学意义(P均>0.05);与NC组相比,sar-1A siRNA组Min6细胞中sar-1A mRNA、sec31蛋白及胰岛素原和X-box蛋白表达均降低(P均<0.05),p-eIF2α、CHOP、XBP1、IRE1蛋白表达均升高(P均<0.05)。与空白血清组相比,5%、10%、20%黄连化浊胶囊组及罗格列酮组Min6细胞中sar-1A mRNA表达均升高(P均<0.05),5%、10%、20%黄连化浊胶囊组Min6细胞中sar-1A mRNA表达逐渐升高,各组间差异均有统计学意义(P均<0.05)。空白血清组与5%黄连化浊胶囊组Min6细胞中sec31蛋白及胰岛素原、p-eIF2α、CHOP、XBP1、IRE1、X-box蛋白表达差异均无统计学意义(P均>0.05);与5%黄连化浊胶囊组相比,10%、20%黄连化浊胶囊组及罗格列酮组Min6细胞中sec31蛋白和胰岛素原表达均增加(P均<0.05),p-eIF2α、CHOP、XBP1、IRE1、X-box蛋白表达均降低(P均<0.05)。结论sar-1A敲除后胰岛β细胞存在COP-Ⅱ功能障碍,黄连化浊胶囊能够降低sar-1A敲除后胰岛β细胞的内质网应激水平。
Objective To investigate the intervention effects of Huanglian Huazhuo capsule on endoplasmic reticulum stress(ERS)of islet β cells with coat protein Ⅱ(COP-Ⅱ)dysfunction induced by secretion associated Ras related GTPase 1A(sar-1A)knockout.Methods Mouse islet β cells Min6 were divided into 3 groups:blank group(Min6 cells without any intervention),negative control(NC)group(Min6 cells were transfected with sar-1A-NC-small interfering RNA[siRNA]),and sar-1A siRNA group(Min6 cells were transfected with sar-1A siRNA).Min6 cells transfected with sar-1A siRNA were collected and divided into blank serum group(cells cultured in medium with 10% blank serum),5%,10%,and 20% Huanglian Huazhuo capsule groups(cells cultured in medium with 5%,10%,and 20%serum containing Huanglian Huazhuo capsule),and rosiglitazone group(cells cultured in medium with 10% serum containing rosiglitazone).The expression of sar-1A mRNA in Min6 cells was detected by quantitative polymerase chain reaction,the expression of sec31 protein was detected by immunofluorescence,and the expression of proinsulin,phosphorylated eukaryotic translation initiation factor 2α(p-eIF2α),C/EBP homologous protein(CHOP),X-box binding protein 1(XBP1),inosital-requiring enzyme 1(IRE1),and X-box proteins was detected by Western blotting.Results There were no significant differences in sar-1A mRNA,sec31,proinsulin,p-eIF2α,CHOP,XBP1,IRE1,or X-box protein between the blank group and the NC group(all P>0.05).Compared with the NC group,the sar-1A siRNA group showed decreased expression of sar-1A mRNA,sec31 protein,proinsulin,and X-box protein(all P<0.05),and increased expression of p-eIF2α,CHOP,XBP1,and IRE1 proteins(all P<0.05).Compared with the blank serum group,the sar-1A mRNA expression in the 5%,10%,and 20% Huanglian Huazhuo capsule groups and the rosiglitazone group was significantly increased(all P<0.05).The expression of sar-1A mRNA was gradually increased in the 5%,10%,and 20% Huanglian Huazhuo capsule groups(all P<0.05).There was no significant difference in the expression of sec31,proinsulin,p-eIF2α,CHOP,XBP1,IRE1,or X-box protein between the blank serum group and the 5% Huanglian Huazhuo capsule group(all P>0.05).Compared with the 5% Huanglian Huazhuo capsule group,sec31 protein and proinsulin in Min6 cells of 10%,20% Huanglian Huazhuo capsule groups and rosiglitazone group were significantly increased(all P<0.05),and p-eIF2α,CHOP,XBP1,IRE1,and X-box proteins were significantly decreased(all P<0.05).Conclusion COP-Ⅱ dysfunction is found in the islet β cells after sar-1A knockdown,and Huanglian Huazhuo capsule can reduce ERS of islet β cells after sar-1A knockdown.