健脾化痰祛瘀法对肥胖2型糖尿病模型糖脂代谢及IRS-1/PI3K/Akt2信号通路的影响

Stasis-Removing Method on Glucose and Lipid Metabolism and the IRS-1/PI3K/Akt2 Signaling Pathway in an Obese Type 2 Diabetes Mellitus Model

目的:探讨健脾化痰祛瘀法对肥胖2型糖尿病(Type 2 diabetes mellitus,T2DM)大鼠的糖脂代谢作用及其机制。方法:通过高脂饮食联合小剂量链佐菌素诱导的T2DM大鼠,分组对照组、模型组、健脾化痰祛瘀方低剂量组、健脾化痰祛瘀方中剂量组、健脾化痰祛瘀方高剂量组。ELISA试剂盒检测血清中促炎细胞因子、生化指标和肝脏中与糖代谢相关的关键酶的活性。qPCR检测胰岛素信号通路关键靶点的mRNA水平。采用Western blot法检测肝脏中磷脂酰肌醇3-激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶B(Akt)、磷酸化Akt(p-Akt)和葡萄糖转运蛋白2(Glut2)的表达。结果:与对照组相比,模型组大鼠血清空腹血糖(FBG)、空腹胰岛素(FINS)、总胆固醇(TC)、甘油三酯(TG)、游离脂肪酸(FFA)、低密度脂蛋白胆固醇(LDL-C)水平显著升高,高密度脂蛋白胆固醇(HDL-C)水平显著降低。而与模型组相比,健脾化痰祛瘀方治疗显著降低了FINS、TG、TC、LDL-C和FFA水平,降低HDL-C水平。此外,与对照组相比,模型组血清CRP、IFN-γ、TNF-α、IL-6、IL-1β、NO水平明显升高。然而,与模型组相比,健脾化痰祛瘀方治疗后,这些促炎细胞因子明显下降。与对照组大鼠相比,模型组大鼠中PEPCK、FBPase、G6Pase和GP的活性增加,但通过健脾化痰祛瘀方治疗后这些酶的活性明显降低。此外,与对照组大鼠相比,模型组大鼠中IRS-1、p-PI3K、p-Akt2的表达增加,但通过健脾化痰祛瘀方治疗后IRS-1、p-PI3K、p-Akt2的表达明显降低。结论:健脾化痰祛瘀法可能通过激活IRS-1/PI3K/Akt2信号通路改善T2DM大鼠的糖脂代谢。

Objective:To explore the effects and mechanisms of the spleen-fortifying,phlegm-resolving,and stasis-removing method on glucose and lipid metabolism in rats with type 2 diabetes mellitus(T2DM).Methods:T2DM was induced in rats using a high-fat diet combined with a low dose of streptozotocin.The rats were divided into control,model,and three treatment groups with low,medium,and high doses of the spleen-fortifying,phlegm-resolving,and stasis-removing formula.Serum inflammatory cytokines,biochemical indicators,and key enzyme activities related to glucose metabolism in the liver were measured using ELISA kits.mRNA levels of key insulin signaling pathway targets were detected by qPCR.Western blotting was used to assess the expression of phosphatidylinositol 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(Akt),phosphorylated Akt(p-Akt),and glucose transporter 2(Glut2)in the liver.Results:Compared with the control group,the model group had significantly elevated serum levels of fasting blood glucose(FBG),fasting insulin(FINS),total cholesterol(TC),triglycerides(TG),free fatty acids(FFA),and low-density lipoprotein cholesterol(LDL-C),and significantly reduced high-density lipoprotein cholesterol(HDL-C).The treatment with the spleen-fortifying,phlegm-resolving,and stasis-removing formula significantly reduced FINS,TG,TC,LDL-C,and FFA levels,while increasing HDL-C levels compared with the model group.Additionally,serum levels of CRP,IFN-γ,TNF-α,IL-6,IL-1β,and NO were significantly higher in the model group compared to the control group,but were significantly reduced after treatment.The activities of PEPCK,FBPase,G6Pase,and GP enzymes were elevated in the model group but decreased significantly following treatment.Moreover,the expression of IRS-1,p-PI3K,and p-Akt2 was increased in the model group but significantly reduced with the treatment.Conclusion:The spleen-fortifying,phlegm-resolving,and stasis-removing method may improve glucose and lipid metabolism in T2DM rats by activating the IRS-1/PI3K/Akt2 signaling pathway.