参苓白术散对肥胖2型糖尿病ZDF大鼠胰岛β细胞去分化的作用  

Effects of Shenling Baizhu Powder on dedifferentiation of isletβcells in ZDF rats of obesity type 2 diabetes mellitus

作　　者：罗金童 张栎婧 战丽彬[2,3] 杭天怡 LUO Jintong;ZHANG Lijing;ZHAN Libin;HANG Tianyi(School of Chinese Medicine&School of Integrated Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210000,China;Center for Innovative Engineering Technology in Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)

机构地区：[1]南京中医药大学中医学院·中西医结合学院,南京210000 [2]辽宁中医药大学中医药创新工程技术中心,沈阳110847 [3]辽宁中医药大学中医脏象理论及应用教育部重点实验室,沈阳110847

出　　处：《中华中医药杂志》2023年第12期6034-6039,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金重点项目(No.81730111),国家中医药领军人才岐黄学者支持项目(No.国中医药人教函【2022】6号);2021年江苏省研究生培养创新工程研究生科研与实践创新项目(No.KYCX21_1704)。

摘　　要：目的:探讨参苓白术散对肥胖2型糖尿病(T2DM) ZDF大鼠胰岛β细胞去分化的作用。方法:10只LZ大鼠为对照组,20只ZDF大鼠随机分为模型组以及参苓白术散治疗组。从5周龄开始对应灌胃超纯水及24 g/kg的参苓白术散水煎液,前期研究证实该剂量疗效及安全性最佳。连续干预至9周龄,检测血清游离脂肪酸(FFA)。通过对照组、模型组、参苓白术散治疗组3组的胰腺组织苏木素-伊红染色,观察胰岛的形态损伤情况。通过免疫荧光实验、蛋白免疫印迹实验、实时荧光定量PCR实验,观察参苓白术散对胰岛中胰岛素和胰高血糖素、叉头转录因子O1(FoxO1)、胰腺十二指肠同源盒-1(Pdx1)、同源盒基因NKx6.1(Nkx6.1)、肌腱膜纤维肉瘤癌基因同源物A(MafA)、八聚体结合蛋白-4(Oct4)、蛋白Nanog、神经元素3(Ngn3)表达的影响,以评价参苓白术散对自发性糖尿病ZDF大鼠胰岛β细胞去分化的影响。结果:9周龄时,与LZ组比较,ZDF组大鼠血清FFA水平升高(P<0.01);在胰腺中,同一视野下,ZDF组胰岛较LZ组形态不规则,边缘不整齐,界限不清晰,胰岛体积略增大。胰岛中胰岛素表达减少(P<0.01),胰高血糖素表达增多(P<0.01)。在胰腺组织中,FoxO1、MafA、Pdx1、Nkx6.1的蛋白及mRNA表达明显降低(P<0.05),Oct4、Nanog蛋白及mRNA、Ngn3蛋白的表达显著升高(P<0.05)。参苓白术散干预后,ZDF大鼠血清FFA水平下降(P<0.01),胰岛形态改善,界限清晰,体积恢复正常。胰岛中胰岛素表达增多(P<0.05),胰高血糖素表达减少(P<0.01)。胰腺中FoxO1、Pdx1、Nkx6.1蛋白及mRNA的表达上调(P<0.05),Oct4、Nanog蛋白及mRNA、Ngn3蛋白的表达下调(P<0.05)。结论:参苓白术散可以降低ZDF大鼠脂毒性风险,改善胰岛β细胞去分化,起到治疗肥胖T2DM的效果。Objective:To investigate the effect of Shenling Baizhu Powder on the dedifferentiation of isletβcells in Zucker diabetic fatty(ZDF)rats with obesity type 2 diabetes mellitus(T2DM).Methods:Ten Lean Zucker(LZ)rats were used as control group,and 20 ZDF rats were randomly divided into model group and Shenling Baizhu Powder treatment group.Superfine water and 24 g/kg water decoction of Shenling Baizhu Powder were administered from 5 weeks old.Previous studies confirmed that this dose had the best curative effect and safety.After continuous intervention until 9 weeks of age,serum free fatty acid(FFA)was detected.The pancreatic islets were observed by hematoxylin-eosin staining in three groups:control group,model group and Shenling Baizhu Powder treatment group.Through immunofluorescence experiment,Western blotting experiment and real-time fuorescence quantitative PCR experiment,the effects of Shenling Baizhu Powder on insulin and glucagon,fork head transcription factor O1(FoxO1),pancreatic duodenal homeodomain-1(Pdx1),NKx6.1(Nkx6.1),v-maf musculoaponeurotic fibrosarcoma oncogene homology A(MafA),octamer-binding transcription factor 4(Oct4),Nanog,Neurogenin-3(Ngn3)were observed to evaluate the effect of Shenling Baizhu Powder on the dedifferentiation of isletβcells in ZDF rats with obesity T2DM.Results:At the age of 9 weeks,compared with LZ group,the serum FFA level in ZDF group increased(P<0.01).In the pancreas,under the same visual field,the islets in ZDF group were irregular in shape,irregular in edge and unclear in boundary,and the islet volume was slightly larger than that in LZ group.The expression of insulin decreased and glucagon(P<0.01)increased in pancreatic islets.The protein and mRNA expression of FoxO1,MafA,Pdx1,Nkx6.1 decreased significantly(P<0.05),while the protein and mRNA expression of Oct4,Nanog,Ngn3 increased significantly(P<0.05).Among them,there was no statistical difference in mRNA expression of Ngn3,but there was an obvious trend.After intervention,the serum FFA level of ZDF rats decreased(P<0.

关 键 词：肥胖2型糖尿病 胰岛β细胞去分化 参苓白术散 脂毒性 

分 类 号：R285.5[医药卫生—中药学]