苍附导痰汤联合二甲双胍对肥胖型PCOS-IR大鼠内分泌激素和炎性因子表达的影响

Effects of Cangfu Daotan Decoction Combined with Metformin on the Expression of Endocrine Hormones and Inflammatory Factors in Obese PCOS-IR Rats

目的:探讨苍附导痰汤与二甲双胍联合用药对肥胖型多囊卵巢综合征联合胰岛素抵抗(PCOS-IR)模型大鼠内分泌激素和炎性因子的影响。方法:采用来曲唑联合高脂饮食建立肥胖型PCOS-IR大鼠模型,并采用快速革兰氏染色法观察动情周期变化,评价模型是否成功。挑选造模成功的大鼠随机分为模型对照组、二甲双胍135 mg/kg组、苍附导痰汤57.96 g/kg组和苍附导痰汤57.96 g/kg+二甲双胍135 mg/kg组,每组各8只。正常对照组和模型对照组大鼠给予等体积纯净水,其余各组分别给予相应药物干预,连续21 d。采用苏木素-伊红(HE)染色观察卵巢组织病理学改变;血液生化仪测定空腹血糖(FBG)变化;酶联免疫吸附(ELISA)法测定卵泡刺激素(FSH)、促黄体生成素(LH)、总睾酮(T)、空腹胰岛素(FINS)和血清炎症因子[肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)]水平;并计算胰岛素抵抗指数HOMA-IR;实时荧光定量PCR测定卵巢Tnf-α和Il-1β mRNA变化。结果:与正常对照组比较,模型对照组大鼠动情周期紊乱,卵巢多囊性改变明显,血清FSH、LH和T分泌显著增加,FINS水平和HOMA-IR明显升高,血清TNF-α、IL-1β水平和卵巢Tnf-α、Il-1β mRNA表达均明显增加(P<0.05或P<0.01);与模型对照组比较,苍附导痰汤57.96 g/kg+二甲双胍135 mg/kg组大鼠卵巢多囊性得到改善,性激素FSH、LH、T水平减少,FINS水平和HOMA-IR均下降,血清TNF-α、IL-1β水平和卵巢Tnf-α、Il-1β mRNA表达均明显下降(P<0.05或P<0.01)。结论:苍附导痰汤与二甲双胍联合用药能有效调节性激素分泌,降低胰岛素抵抗和炎性因子水平,改善肥胖型PCOS-IR大鼠卵巢生殖功能。

Objective:To investigate the effects of Cangfu Daotan Decoction(CFDT) combined with metformin on endocrine hormones and inflammatory factors in obese polycystic ovary syndrome combined with insulin resistance(PCOS-IR) model rats. Methods: Letrozole combined with a high-fat diet was used to establish the PCOS-IR model in rats. Gram staining was used to observe changes in the estrous cycle to evaluate the established model. PCOS-IR model rats were randomly divided into a model group, a metformin(135 mg/kg) group, a CFDT(57.96 g/kg) group, and a CFDT(57.96 g/kg) + metformin(135 mg/kg) group, with eight rats in each group. Rats in the normal group and the model group received pure water at an equal volume, and those in the groups with drug intervention were treated correspondingly for 21 consecutive days. Hematoxylin-eosin(HE) staining was used to observe histopathological changes of the ovarian tissues. The fasting blood glucose(FBG) was determined by a blood chemistry analyzer. Enzyme-linked immunosorbent assay(ELISA) was used to determine the levels of sex hormones [follicle stimulating hormone(FSH), luteinizing hormone(LH), total testosterone(T)], fasting insulin(FINS), and serum inflammatory factors [tumor necrosis factor-a(TNF-a) and interleukin-1β(IL-1β)]. Also the IR index HOMA-IR was calculated. Real-time fluorescence-based quantitative PCR was used to detect the changes in ovarian TNF-a and IL-1β mRNA. Results: Compared with the normal group, the model group showed disordered estrous cycle, manifest ovarian polycystic changes, increased serum levels of FSH, LH, and T secretion, elevated levels of FINS and HOMA-IR, and up regulated serum levels of TNF-a and IL-1β and mRNA expression of ovarian TNF-α and Il-1β(P<0.05 or P<0.01). Compared with the model group, the CFDT + metformin group showed improved polycystic ovary, decreased levels of FSH, LH, and T, declining FINS and HOMA-IR, and down regulated serum levels of TNF-a and IL-1β and ovarian levels of TNF-α and IL-1β(P<0.05 or P<0.01). Conclusion: The combination of CFDT and metformin can effectively regulate the secretion of sex hormones, reduce IR and inflammatory factor levels, and improve the ovarian reproductive function on obese PCOS-IR rats.