摘要
目的:基于胰岛素受体底物-1(IRS-1)/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)通路研究二冬汤对2型糖尿病(T2DM)小鼠肝脏糖脂代谢紊乱的治疗作用及其作用机制。方法:采用高脂高糖饲料联合腹腔注射链脲佐菌素(STZ)建立T2DM小鼠模型。将造模成功的小鼠分为模型组,二甲双胍组及二冬汤高、中、低剂量组。二甲双胍组和二冬汤高、中、低剂量组灌胃剂量分别为0.15、22.64、11.32、5.66 g·kg^(-1),另设空白组,模型组和空白组灌胃等量生理盐水。灌胃给药8周后,测量小鼠空腹血糖(FBG)、体质量、肝脏质量、肝脏系数、进行口服葡萄糖耐量试验(OGTT)。使用酶联免疫吸附测定法(ELISA)检测血清胰岛素(FINS)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、活性氧(ROS),计算胰岛素抵抗指数(HOMA-IR)。检测血清中总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)。苏木素-伊红(HE)染色观察肝脏组织病理损伤。蛋白免疫印迹法(Western blot)检测肝脏组织IRS-1、PI3K、Akt、磷酸化(p)-Akt的蛋白表达量。结果:与空白组比较,模型组FBG、FINS、HOMA-IR、IL-6、TNF-α、ROS、TC、TG、LDL-C、肝脏质量、肝脏系数显著升高(P<0.01);HDL-C,IRS-1、PI3K、Akt、p-Akt/Akt蛋白,体质量显著减少(P<0.01),肝脏病理组织中肝小叶形态不清晰,肝细胞排列紊乱,肝细胞核增大,核膜不清晰,脂肪变性明显,炎症细胞浸润。与模型组比较,二甲双胍组、二冬汤高、中、低剂量组小鼠FBG、FINS、HOMA-IR、IL-6、TNF-α、ROS、TG、TC、LDL-C含量明显降低(P<0.05,P<0.01);二甲双胍组、二冬汤高、中剂量组小鼠HDL-C,IRS-1、PI3K、Akt、p-Akt/Akt蛋白表达明显增加(P<0.05,P<0.01)。二甲双胍组、二冬汤高、中、低剂量组小鼠p-Akt/Akt表达显著增加(P<0.01),各给药组小鼠肝脏病理组织中脂肪变性和炎症细胞浸润得到明显改善。结论:二冬汤可以有效改善T2DM小鼠的糖脂代谢、减轻炎症反应,缓解氧化应激,其机制可能与IRS-1/PI3K/Akt信号通路激活有关。
Objective:To investigate the therapeutic effects of Erdong Tang on hepatic glycolipid metabolism disorders in type 2 diabetes mellitus(T2DM)mice and explore the mechanism on the basis of the insulin receptor substrate 1(IRS-1)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)pathway.Methods:A T2DM mouse model was established by high-fat and high-sugar feed combined with intraperitoneal injection of streptozotocin(STZ).Mice successfully modeled were divided into a model group,a metformin group,and high-dose,medium-dose,and low-dose groups of Erdong Tang.The gavage doses for the metformin group and the high-dose,medium-dose,and low-dose groups of Erdong Tang were 0.15,22.64,11.32,5.66 g·kg^(-1),respectively.At the same time,a blank group was set up,and the model and blank groups were administered with equal amounts of saline by gavage.After eight weeks of gavage administration,mice were measured for fasting blood glucose(FBG),body mass,liver weight,liver coefficient,and an oral glucose tolerance test(OGTT)was conducted.Serum insulin(FINS),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and reactive oxygen species(ROS)were measured by enzyme-linked immunosorbent assay(ELISA),and insulin resistance index(HOMA-IR)was calculated.Total cholesterol(TC),triglycerides(TG),highdensity lipoprotein(HDL),and low-density lipoprotein(LDL)were detected.Hematoxylin-eosin(HE)staining was used to observe the histopathological damage of the liver.Western blot was used to detect the protein expression of IRS-1,PI3K,Akt,and p-Akt in liver tissue.Results:Compared with those in the blank group,FBG,FINS,HOMA-IR,IL-6,TNF-α,ROS,TC,TG,LDL-C,liver weight,and liver coefficient were significantly increased in the model group(P<0.01),and HDL-C,IRS-1,PI3K,Akt,p-Akt/Akt protein,and body mass were significantly reduced(P<0.0l).The pathological tissue in the liver showed unclear morphology of hepatic lobules,disordered arrangement of hepatocytes,enlarged nuclei of hepatocytes,unclear nuclear membrane,obvious fatty degeneration,and infiltration of inflammatory cells.Compared with that in the model group,the FBG,FINS,HOMA-IR,IL-6,TNF-α,ROS,TG,TC,and LDL-C content of mice was significantly reduced in the metformin group and the high-dose,medium-dose,and low-dose groups of Erdong Tang(P<0.05,P<0.01).The expression of HDL-C,IRS-1,PI3K,and Akt protein was significantly increased in the metformin group and the high-dose and medium-dose groups of Erdong Tang(P<0.05,P<0.01).The expression of p-Akt/Akt in mice was significantly increased in the metformin group and the high-dose,mediumdose,and low-dose groups of Erdong Tang(P<0.01),and fatty degeneration and inflammatory cell infiltration were significantly improved in the pathological tissues of the mouse liver in each dosing group.Conclusion:Erdong Tang ameliorates glycolipid metabolism,inflammatory responses,and oxidative stress in T2DM mice,potentially by activating the IRS-1/PI3K/Akt signaling pathway.
作者
李浩
姜立娟
张文风
LI Hao;JIANG Lijuan;ZHANG Wenfeng(College of Basic Medical Sciences,Changchun University of Chinese Medicine,Changchun 130117,China)
出处
《中国实验方剂学杂志》
北大核心
2025年第23期21-29,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
吉林省卫生健康科技能力提升项目(2023JC039)
吉林省教育厅科学技术研究项目(JJKH20241056KJ)
吉林省中医药管理局项目-张文风名老中医专家传承工作室项目(吉中医药发[2023]56号)。
