补肾化痰方对多囊卵巢综合征胰岛素抵抗Akt通路调控的实验研究

Effect of Bushen Huatan Recipe on the Akt Signal Pathway in Polycystic Ovarian Syndrome Model Rats with Insulin Resistance:an Experimental Research

目的探讨补肾化痰方对多囊卵巢综合征(polycystic ovarian syndrome,PCOS)模型大鼠卵巢内胰岛素信号传导分子的调控。方法 Wistar雌鼠50只,随机分为溶媒对照组、模型组、中药低剂量组(5.406 g/kg)、中药中剂量组(10.812 g/kg)、中药高剂量组(21.624 g/kg),每组10只。测定大鼠空腹血糖及胰岛素水平,计算稳态模型胰岛素抵抗指数(HOMA-IR);RT-PCR技术检测糖原合成酶激酶-3β(GSK-3β)、葡萄糖转运蛋白4(GLUT-4)、胰岛素受体底物-1(IRS-1)以及过氧化物酶增殖体激活受体γ(PPAR-γ)表达,Western blot法检测卵巢内胰岛素信号传导分子蛋白激酶B(Akt)表达。结果与溶媒对照组比较,模型组HOMA-IR及PPAR-γmRNA表达明显升高,卵巢GSK-3β、GLUT-4、IRS-1 mRNA表达及Akt、p-Akt蛋白表达均明显降低,差异均有统计学意义(P<0.05)。与模型组比较,中药高剂量组HOMA-IR明显降低,GSK-3β、GLUT-4、IRS-1 mRNA表达及Akt蛋白表达均显著升高,p-Akt蛋白表达尤为显著,差异均有统计学意义(P<0.05,P<0.01);中药低、中剂量组GSK-3β、GLUT-4 mRNA显著升高,PPAR-γmRNA明显降低,差异有统计学意义(P<0.05);中药低剂量组p-Akt蛋白表达明显升高,差异有统计学意义(P<0.05)。结论 PCOS模型鼠存在胰岛素抵抗,并存在卵巢组织信号通路传导异常。补肾化痰方能够改善PCOS大鼠胰岛素抵抗,与胰岛素信号传导分子蛋白表达的调控有关。

Objective To observe the effect of Bushen Huatan Recipe （BHR） on the Akt signal pathway in polycystic ovarian syndrome （PCOS） model rats with insulin resistance （IR）. Methods Fifty Wistar female PCOS rats were randomly and equally divided into 5 groups, i.e., the control group, the model group, the low dose BHR group （5.406 g/kg）, the medium dose BHR group （10.812 g/kg）, and the high dose BHR group （21. 624 g/kg）, 10 in each group. Levels of fasting blood glucose （FBG） and in- sulin were detected to calculate homeostasis model assessment of insulin resistance （HOMA-IR）. The expression of insulin receptor substrate 1 （ IRS-1 ）, glycogen synthetase kinase- 3β （GSK- 3β）, glucose transporter 4 （GLUT-4）, and peroxisome proliferator activated receptor （PPAR-~/） mRNA were detected by RT-PCR. The expression of insulin signal transduction molecular kinase B （Akt） was detected by Western blot. Results Compared with the control group, HOMA-IR and the mRNA expression of PPAR--y mRNA significantly increased, the mRNA expression of GSK-3β, GLUT-4, and IRS-1, protein expression of Akt and p-Akt significantly decreased in the model group （P 〈0.05）. Compared with the model group, HOMA-IR significantly decreased, the mRNA expression of GSK-3β, GLUT-4, IRS-1, and Akt protein sig- nificantly increased in the high dose BHR group （P 〈0.05, P 〈0.01 ）. The mRNA expression of p-Akt pro- tein increased more obviously （P 〈0.05, P 〈0.01 ）. mRNA expression of GSK- 3β and GLUT-4 signifi- cantly increased （P 〈0.05）, while the mRNA expression of PPAR-h, significantly decreased in the lowand middle BHR groups （P 〈0.05）. The expression of p-Akt significantly increased in the low dose BHR group （P 〈0.05）. Conclusions IR and abnormal insulin signal pathway existed in PCOS model rats. BHR could improve IR of PCOS rats, which was correlated with regulating protein expression of insulin signal transduction molecules.