基于内质网应激探讨黄连-大黄药对对2型糖尿病GK大鼠胰岛β细胞功能的影响  

Effect of COPTIDIS RHIZOMA-RHEI RADIX ET RHIZOMA on Pancreatic β-Cell in GK Rats Based on Endoplasmic Reticulum Stress

作　　者：李振华 陈源 岳仁宋[3] 张博荀 杨茂艺 LI Zhenhua;CHEN Yuan;YUE Rensong;ZHANG Boxun;YANG Maoyi(Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000;Shaanxi University of Chinese Medicine,Xi'an 712046;Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072)

机构地区：[1]陕西中医药大学附属医院,咸阳日712000 [2]陕西中医药大学,西安712046 [3]成都中医药大学附属医院,成都610072

出　　处：《中药药理与临床》2024年第7期31-36,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金(编号:81774279);陕西省自然科学基础研究计划-一般(面上)项目(编号:2022JM-495);陕西省中医药管理局基础类项目(编号:2021-ZZ-JC004);陕西中医药大学国家基金培育项目(编号:2021GP22);陕西市科技局重点研发计划-社发项目(编号:L2022ZDYFSF003)。

摘　　要：目的:基于内质网应激途径探讨黄连-大黄药对改善2型糖尿病(T2DM)GK大鼠胰岛β细胞功能的作用机制。方法:将符合成模标准的自发性T2DM GK大鼠随机分为模型对照组、二甲双胍0.1 g/kg组、黄连-大黄药对2.36、4.72 g/kg组,并取正常Wistar大鼠设为正常对照组,每组各6只,干预8 w后采用HE染色观察大鼠胰腺病理形态学变化;ELISA法检测胰腺组织胰岛素(INS)水平;实时荧光定量PCR法检测胰腺组织胰十二指肠同源盒因子1(Pdx1)、葡萄糖调节蛋白78(Grp78)mRNA表达;IHC法检测胰腺组织葡萄糖转运蛋白2(GLUT2)、GRP78蛋白表达;Western blot法检测胰腺组织磷酸化肌醇依赖性激酶1α(p-IRE1α)、磷酸化真核翻译起始因子2α(p-EIF2α)蛋白表达。结果:与正常对照组比较,模型对照组大鼠胰岛形态结构紊乱,胰岛细胞数目减少,伴有炎性细胞浸润,胰腺组织中INS水平、Pdx1 mRNA及GLUT2蛋白表达均显著下调(P<0.01),Grp78 mRNA与蛋白以及p-IRE1α、p-EIF2α蛋白表达均显著上调(P<0.01);与模型对照组比较,黄连-大黄药对2.36、4.72 g/kg组大鼠胰岛形态结构改善,胰腺组织中INS水平、Pdx1 mRNA及GLUT2蛋白表达明显上调(P<0.05),Grp78 mRNA与蛋白以及p-IRE1α、p-EIF2α蛋白表达均下调(P<0.05)。结论:黄连-大黄药对可能通过抑制内质网应激改善胰岛β细胞功能,发挥T2DM治疗作用。Objective:To investigate the mechanism underlying the improvement of pancreatic β-cell function in Goto-Kakizaki(GK) rats by COPTIDIS RHIZOMA-RHEI RADIX ET RHIZOMA based on the endoplasmic reticulum stress pathway. Methods:The spontaneous GK rats that met the modeling criteria were randomly divided into model control group, metformin(0.1 g/kg) group, and COPTIDIS RHIZOMA-RHEI RADIX ET RHIZOMA(2.36 g/kg group and 4.72 g/kg) groups, and normal Wistar rats were selected as normal control group, with 6 in each group. After 8 weeks of intervention, HE staining was performed to observe the pathological changes of pancreas in rats. ELISA was used to measure insulin(INS) in pancreatic tissue. Real-time PCR was used to detect the mRNA expressions of pancreatic and duodenal homeobox 1(Pdx1) and glucose regulatory protein 78(Grp78) in pancreatic tissue. The expressions of glucose transporter protein 2(GLUT2) and GRP78 proteins in pancreatic tissue were detected by IHC method, while those of phosphorylated inositol-requiring enzyme1α(p-IRE1α) and phosphorylation of eukaryotic initiation factor 2α(p-EIF2α) proteins were detected by Western blot. Results:Compared with the normal control group, the model control group presented disordered structure of islets and reduced number of islet cells with inflammatory cell infiltration. Additionally, the INS level and Pdx1 mRNA and GLUT2 protein expressions in pancreatic tissue were decreased(P<0.01),while Grp78 mRNA and protein expressions and the expressions of p-IRE1α and p-EIF2α proteins were up-regulated(P<0.01). Compared with the model control group, the COPTIDIS RHIZOMA-RHEI RADIX ET RHIZOMA(2.36 g/kg group and 4.72 g/kg) groups improved the morphological structure of islets in rats, and the INS level and Pdx1 mRNA and GLUT2 protein expressions in pancreatic tissue were elevated(P<0.05),while Grp78 mRNA and protein expressions and the expressions of p-IRE1α and p-EIF2α proteins were decreased(P<0.05),with the concentration of 2.36 g/kg exhibiting better effect. Conclusio

关 键 词：黄连-大黄药对 2型糖尿病 内质网应激 胰岛Β细胞 

分 类 号：R285.5[医药卫生—中药学]