健脾清化方对糖尿病模型大鼠胰岛素敏感性及骨骼肌p38MAPK信号通路的影响  

Effect of Jianpi Qinghua Recipe on Insulin Sensitivity and p38MAPK Signaling of skeletal muscle in diabetic rats

作　　者：肖岩岩 陈清光[1] 韩煦 徐隽斐[1] XIAO Yanyan;CHEN Qingguang;HAN Xu(Endocrinology Department of Shuguang Hospital,Shanghai Municipality,Shanghai 200021,China)

机构地区：[1]上海中医药大学附属曙光医院内分泌科,上海200021

出　　处：《四川中医》2023年第7期57-62,共6页Journal of Sichuan of Traditional Chinese Medicine

基　　金：上海中医药大学产业发展中心医养结合科创项目(编号:YYKC-2021-01-145);国家自然科学基金委员会面上项目(编号:82074381);国家自然科学基金委员会面上项目(编号:81874434);上海市科委科研计划项目(编号:21S21900700);华东片区及市级中医专科专病联盟建设(编号:ZY(2021-2023)-0302)。

摘　　要：目的:观察健脾清化方对糖尿病模型大鼠胰岛素敏感性的影响,探讨p38丝裂原活化的蛋白激酶(p38MAPK)信号通路在其中的作用机制。方法:利用高脂高糖饲料协同小剂量链脲佐菌素来诱导建立Wistar大鼠糖尿病模型,按照随机数字表法分为模型组(CON组)、健脾清化方组(JPQ组)、吡格列酮组(PIO组),每组8只,另取同批次8只大鼠为空白对照组(NOR组),以普通饲料喂养。正葡萄糖高胰岛素钳夹实验和腹腔胰岛素耐量试验(Intraperitoneal Insulin Tolerance Test,IPITT)观察健脾清化方对大鼠胰岛素敏感性的影响。反转录聚合酶链式反应法(Reverse Transcription-Polymerase Chain Reaction,Rt-PCR)观察健脾清化方对大鼠骨骼肌p38MAPK通路的关键信号分子(MKK3、MKK6、p38MAPK)及葡萄糖转运体4(glucose transporter type-4,Glut-4)基因表达水平的影响,蛋白质免疫印迹法(Western blot)观察健脾清化方对大鼠骨骼肌p38MAPK通路的关键信号分子(MKK3、MKK6、p38MAPK、p-p38MAPK)及Glut-4蛋白表达水平的影响。结果:与模型组比较,健脾清化方能提高葡萄糖输注率(P<0.01),降低胰岛素曲线下面积(P<0.01),有效提高模型大鼠胰岛素敏感性。与模型组相比,健脾清化方能抑制MKK3的基因和蛋白表达(P<0.05),降低骨骼肌p38MAPK的基因和蛋白表达(P<0.05)及磷酸化,提高Glut-4基因和蛋白的表达(P<0.05)。结论:健脾清化方可能通过抑制p38MAPK信号通路,促进Glut-4基因和蛋白表达,改善糖尿病模型大鼠胰岛素抵抗。Objective:To observe the effect of Jianpi Qinghua Recipe on insulin sensitivity in diabetic rats,explore the mechanism of p38MAPK signal pathway.Methods:The diabetic model of Wistar rats was induced by low dose streptozotocin combined with high fat and high sugar diet,which were divided into model group(CON group),Jianpi Qinghua recipe group(JPQ group),pioglitazone group(PIO group)according to randomized digital table,8rats in each group,and 8rats of the same batch were taken as the blank control group(NOR group),which were fed with ordinary feed.The effects of Jianpi Qinghua Recipe on insulin sensitivity of rats were observed by euglycemic hyperinsulinemic clamp test and intraperitoneal insulin tolerance test(IPITT).Rt-PCR method was used to observe the effect of Jianpi Qinghua Recipe on the gene expression level of Glut-4molecule and key signal molecules(MKK3、MKK6、p38MAPK)in p38MAPK pathway of rat skeletal muscle,and Western-Blot method was used to observe the effect of Jianpi Qinghua Recipe on the protein expression level of Glut-4molecule and key signal molecules(MKK3、MKK6、p38MAPK、p-p38MAPK)in p38MAPK pathway of rat skeletal muscle.Results:Compared with the model group,Jianpi Qinghua Recipe can improve glucose infusion rates(P<0.01),reduce the area under insulin curve(P<0.01),and effectively improve insulin sensitivity of the model rats.Compared with the model group,Jianpi Qinghua Recipe could reduce the protein and gene expression of MKK3(P<0.05),inhibit the gene and protein expression of MKK3(P<0.05),decrease the gene and protein expression and phosphorylation of p38MAPK in skeletal muscle,and improve the protein and gene expression of Glut-4(P<0.05).Conclusion:Jianpi Qinghua Recipe may alleviate insulin resistance by inhibiting the p38MAPK signaling pathway and improving glut-4gene and protein expression.

关 键 词：糖尿病 健脾清化方 骨骼肌p38MAPK通路 GLUT-4 胰岛素敏感性 

分 类 号：R285.5[医药卫生—中药学]