痰脂消汤调控PCOS-IR模型大鼠卵巢PI3K/Akt途径探讨

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目的:观察痰脂消汤对来曲唑灌胃联合高脂膳食诱导的多囊卵巢综合征胰岛素抵抗(PCOS-IR)模型大鼠卵巢胰岛素信号转导途径的影响。方法:来曲唑灌胃联合高脂膳食喂养SD大鼠建立PCOS-IR模型,大鼠随机分为模型组、痰脂消汤组(35 g·kg~(-1)·d~(-1))、二甲双胍组(200 mg·kg~(-1)·d~(-1)),连续灌胃20 d;另设正常组。实验结束后,腹主动脉采血,测定血清空腹血糖(FPG)和空腹胰岛素(FINS)浓度,并计算胰岛素抵抗指数(HOMA-IR);实时荧光定量PCR(Real-time PCR)检测大鼠卵巢胰岛素受体(INSR),胰岛素受体底物(IRS)-1,酯酰肌醇-3激酶(PI3K),蛋白激酶B(Akt)mRNA表达;蛋白免疫印迹法(Western blot)检测卵巢组织INSR,IRS-1,PI3K,Akt蛋白及其相应磷酸化水平表达。结果:与空白组比较,模型组大鼠空腹胰岛素(FINS),HOMA-IR水平均显著升高(P<0.01),FPG无明显升高;模型组大鼠卵巢INSR,IRS,PI3K,Akt mRNA表达水平明显降低(P<0.05);INSR,p-INSR,IRS,p-IRS,PI3K,p-PI3K,Akt,p-Akt等蛋白表达水平明显降低(P<0.05);INSR,IRS,PI3K及Akt磷酸化比例明显降低(P<0.05)。与模型组比较,中药和二甲双胍治疗后大鼠FINS,HOMA-IR均显著降低(P<0.01);大鼠卵巢INSR,IRS,PI3K,Akt mRNA表达水平均明显升高(P<0.05);INSR,p-INSR,IRS,p-IRS,PI3K,p-PI3K,Akt,p-Akt等蛋白表达水平明显升高(P<0.05);INSR,IRS,PI3K及Akt磷酸化比例明显升高(P<0.05)。结论:PCOS大鼠存在IR,卵巢内PI3K/Akt信号途径转导存在异常。痰脂消汤能够显著改善大鼠胰岛素抵抗,可能与调控胰岛素信号转导通路中关键的蛋白表达有关。 Objective: To observe the effect of Tanzhixiao decoction on ovarian insulin signal transduction pathway in rats with polycystic ovary syndrome and insulin resistance (PCOS-IR) induced by levullalamic and high fat diet. Methods: PCOS-IR model was established by intragastric administration of letrozole and high-fat diet to rats. The rats were randomly divided into model group, phlegm and resolving fat group (35 g · kg -1 d -1 ), Metformin group (200 mg · kg -1 · d -1) for 20 days, and another normal group. At the end of the experiment, the abdominal aorta blood was collected for determination of serum fasting blood glucose (FPG) and fasting insulin (FINS) concentrations, and calculate the insulin resistance index (HOMA-IR); Real-time fluorescence quantitative PCR (INSR), insulin receptor substrate (IRS) -1, PI3K and Akt mRNA were detected by immunohistochemistry. The expressions of INSR, IRS-1, PI3K, Akt proteins and their corresponding phosphorylation levels. Results: Compared with the blank group, the levels of fasting insulin (FINS) and HOMA-IR in the model group were significantly increased (P <0.01), while the FPG did not increase significantly. The mRNA expressions of INSR, IRS, PI3K and Akt The expression of INSR, p-INSR, IRS, p-IRS, PI3K, p-PI3K, Akt and p-Akt were significantly decreased And Akt phosphorylation was significantly decreased (P <0.05). Compared with the model group, the FINS and HOMA-IR of rats treated with Chinese medicine and metformin were significantly decreased (P <0.01); the expression of INSR, IRS, PI3K and Akt mRNA in ovary of rats were significantly increased (P <0.05) , P-INSR, IRS, p-IRS, PI3K, p-PI3K, Akt, p-Akt and other proteins were significantly increased <0.05). CONCLUSION: There is IR in PCOS rats, and abnormal PI3K / Akt signaling pathway in ovary. Tanyaxiao Decoction can significantly improve insulin resistance in rats, which may be related to the regulation of key protein expression in insulin signaling pathway.
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