白虎加人参汤加减方对2型糖尿病大鼠胰岛细胞的保护作用与机制研究  

Protective Effect and Mechanism of Modified Baihu and Renshen Decoction on Islet Cells in Type 2 Diabetic Rats

作　　者：王芳 王力彬 刘清[2] 高天慧 宋小妹[2] WANG Fang;WANG Libin;LIU Qing;GAO Tianhui;SONG Xiaomei(School of Medicine,Shaanxi Energy Institute,Xianyang 712000 Shaanxi,China;Shaanxi University of Chinese Medicine,School of Pharmacy,Xianyang 712046 Shaanxi,China)

机构地区：[1]陕西能源职业技术学院医学院,陕西咸阳712000 [2]陕西中医药大学药学院,陕西咸阳712000

出　　处：《中药新药与临床药理》2022年第9期1189-1196,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：陕西省科技厅自然科学基础研究计划项目(2021IQ-888);陕西省教育厅科研计划项目(21JK0578);陕西能源职业技术学院校级自然科学研究重点课题(2021KYZ01);陕西能源职业技术学院慢性病健康服务创新团队项目。

摘　　要：目的 观察白虎加人参汤加减方对2型糖尿病(T2DM)大鼠胰岛细胞的保护作用,并探讨其作用机制。方法 采用高脂高糖饲料联合注射链脲佐菌素(STZ)的方法制备2型糖尿病大鼠模型,将成模大鼠分为模型组、阳性药组及白虎加人参汤加减方高、低剂量组,连续给药8周。观察大鼠一般状态及测定体质量,分别检测生化指标糖化血红蛋白(HbAlc)、空腹血糖(FBG)、胰岛素(INS),计算胰重比;采用HE染色观察2型糖尿病大鼠胰腺组织病理变化;采用RT-PCR法测定胰腺组织中含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、Bcl-2相关X蛋白(BAX)、B淋巴细胞瘤-2基因(Bcl-2)mRNA水平;采用Western Blot技术检测胰腺组织中Cleaved caspase-3、蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)、胰十二指肠同源盒因子-1(PDX-1)、细胞核中插头转录因子1(n-FoxO1)、葡萄糖激酶(GCK)蛋白表达,免疫组化方法检测n-FoxO1蛋白表达。结果 与模型组相比,白虎加人参汤加减方高、低剂量组能够改善2型糖尿病大鼠的多饮、多尿现象,体质量明显增加(P<0.05),降低2型糖尿病大鼠空腹血糖、糖化血红蛋白、 Caspase-3 mRNA、 Bax mRNA、Cleaved caspase-3和n-FoxO1蛋白水平,升高INS、Bcl-2 mRNA水平和p-AKT/AKT,PDX-1、GCK蛋白水平,增加胰重比,改善胰腺组织病理损伤,差异具有统计学意义(P<0.05)。结论 白虎加人参汤加减方可以减少2型糖尿病大鼠胰岛β细胞凋亡,增加胰重比,改善胰腺组织的病理变化,对2型糖尿病大鼠胰岛细胞具有保护作用,其作用机制可能与调节PI3K/AKT/FoxO1信号通路上的相关蛋白表达有关。Objective To observe the protective effect and mechanism of modified Baihu and Renshen Decoction(MBRD) on islet cells of type 2 diabetic(T2DM) rats. Methods High-fat and high-sugar diet combined with streptozotocin(STZ)injection was used to prepare T2DM rat models. The model rats were divided into model group,positive group,MBRD high-and low-dose groups,which were administrated continuously for 8 weeks. The general state and body weight of the rats were observed. The biochemical indexes including glycated hemoglobin(HbAlc),fasting blood glucose(FBG) and insulin(INS) were detected, and the pancreatic weight/body weight ratio was calculated. HE staining was used to observe the pathological changes of pancreatic tissue in T2DM rats. The mRNA levels of cysteine-containing aspartate proteolytic enzyme 3(Caspase-3),Bcl-2 related X protein(BAX)and B lymphoma-2 gene(Bcl-2) in pancreatic tissue were detected by RT-PCR. The protein expressions of Cleaved caspase-3,phosphorylated protein kinase B(p-AKT),protein kinase B(AKT),pancreatic duodenal homeobox 1(PDX-1),nuclear plug transcription factor 1(n-FoxO1)and glucokinase(GCK)in pancreatic tissue were detected by Western Blot,and the protein expression of n-FoxO1 was detected by immunohistochemistry. Results Compared with model group,MBRD high-and low-dose groups could improve the phenomenon of polydipsia and polyuria,increase body weight significantly(P<0.05), decrease the levels of FBG, HbAlc, Caspase-3 mRNA, Bax mRNA,Cleaved caspase-3 and n-FoxO1 protein in T2DM rats. The levels of INS,Bcl-2 mRNA,p-AKT/AKT,PDX-1 and GCK protein were increased,the pancreatic weight/body weight ratio was increased,the pathological injury of pancreatic tissue was improved, and the differences were statistically significant(P<0.05). Conclusion modified Baihu and Renshen Decoction can reduce the apoptosis of islet β cells in T2DM rats,increase the ratio of pancreatic weight/body weight and improve the pathological changes of pancreatic tissue,and show protective effects on islet cells in T2DM

关 键 词：白虎加人参汤加减方 2型糖尿病(T2DM) 胰岛细胞 PI3K/AKT/FoxO1信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]