补肾化痰法对肥胖型PCOS雌鼠AdipoR1、AMPK的影响及其调控胰岛素抵抗的作用

Effect of Kidney-Nourishing and Phlegm-Removing Method on Expressions of AdipoR1,AMPK and Regulating IR of Obese Rats with PCOS

目的：观察补肾化痰法经APN/AMPK通路调节肥胖型PCOS大鼠胰岛素抵抗的作用及机制。方法：选取7-8周龄SD雌性大鼠,来曲唑联合高脂灌服制备肥胖型PCOS模型,以补肾化痰法治疗,放射免疫法检测大鼠血清FSH、LH、E、P、T,免疫组织化学染色法检测AdipoR1、InR,Western blotting检测p-AMPK蛋白表达水平。结果：（1）与模型对照组比较,各治疗组排卵率显著升高;中药高剂量组、中剂量组大鼠体质量增长率显著低于模型组（P〈0.05）;（2）与模型对照组比较,中药高、中剂量组血清FSH、P、E2含量显著增高（P〈0.05或P〈0.01）,中药中剂量组血清LH显著降低（P〈0.01）,中药高、中剂量组血清T含量降低（P〈0.05）;（3）模型对照组卵巢颗粒细胞AdipoR1、p-AMPK表达显著低于空白对照组（P〈0.01）,而中药高、中剂量组卵巢颗粒细胞AdipoR1表达明显高于模型对照组（P〈0.05或P〈0.01）,中药各剂量组p-AMPK均高于模型对照组（P〈0.01）;（4）与模型对照组比较,中药高、中剂量组FBG、Fins、HOMA-IR、INR显著下降（P〈0.05或P〈0.01）,中药高、中剂量组卵巢组织胰岛素受体表达显著增高（P〈0.05）。结论：肥胖型PCOS雌鼠出现排卵障碍,生殖内分泌紊乱的表现,并伴随全身的糖代谢异常及胰岛素抵抗。补肾化痰法可能经APN/AMPK通路改善肥胖型PCOS的肥胖状态、生殖内分泌紊乱及胰岛素抵抗。

Objective：To observe the regulative effect and mechanism of of kidney-nourishing and phlegm-removing method（KNPR） on insulin resistance（IR） of obese rats with PCOS through the APN/AMPK pathway. Methods：We chose 7-8 weeks old female SD rats,and treated them with the method of KNPR after modeling by Letrozole and high-fat and cholesterol. The serum levels of E2,P,T,FSH and LH were detected by radioimmunoassay. The expressions of AdipoR1 and INR were detected by immunohistochemical staining method. And the expression of p-AMPK was detected by Western blotting. Results：（1） Compared with the model group,the ovulation rate of treatment group was increased significantly which was the same as that of the Diane-35 group,and the weight growth rate of the traditional Chinese medicine high dose group（ HDG） and middle dose group（ MDG）was lower（ P〈0.05）.（2）Compared with the model group,the FSH,P and E2 of HDG and MDG had increased significantly（ P〈0.05 or P〈0.01）. The LH level of MDG decreased significantly（ P〈0.01）. And the T levels of HDG and MDG decreased significantly（ P〈0.05）.（3）The expressions of AdipoR1 and p-AMPK in granular cell（ GC） of the model group were lower than that of the control group（ P〈0.01）. Compared with the model group,the AdipoR1 of HDG and MDG increased（ P〈0.05 or P〈0.01） and the p-AMPK of treatment groups increased（ P〈0.01）.（4）Compared with model group,the FBG,Fins,HOMA-IR and insulin receptor（ INR） of the HDG and MDG decreased significantly（ P〈0.05 or P〈0.01）. The expression of INR was higher in HDG and MDG（ P〈0.05）. Conclusion：The obese rats with PCOS showed ovulation failure,reproductive endocrine disorder and IR. And the method of KNPR could improve these pathological symptoms through APN/AMPK pathway.