目的 建立多囊卵巢综合征（PCOS）大鼠模型，探究六味地黄丸是否通过转化生长因子β1（TGF-β1）/SMAD信号通路对PCOS大鼠内分泌代谢产生影响。方法 将60只SD雌性大鼠随机分为对照组、模型组、二甲双胍（阳性药，50mg·kg^-1）组、六味地黄丸（50mg·kg^-1）组、六味地黄丸（50mg·kg^-1）＋SRI-011381（TGF-β1/SMAD通路激活剂，100ng·kg^-1）组，除对照组外，其余各组每天ig1mg·kg^-1来曲唑（溶于0.5%羧甲基纤维素钠），并喂食高脂饲料，连续15d，构建PCOS大鼠模型；对照组每日ig等体积0.5%羧甲基纤维素钠，并喂食普通饲料。造模完成后，各组ig给药，对照组、模型组分别ig等体积0.9%氯化钠溶液，连续10d。苏木精-伊红（HE）染色法检测各组大鼠卵巢、子宫的病理变化情况；采用酶联免疫试剂盒（ELISA）法检测各组大鼠血清睾酮（T）、黄体生成素（LH）、卵泡刺激素（FSH）水平；采用实时荧光定量PCR（qRT-PCR）法检测各组卵巢组织中TGF-β1、SMAD2、SMAD3mRNA的表达水平，采用Western blotting法检测各组卵巢组织中TGF-β1、SMAD2、SMAD3蛋白表达水平。结果 对照组大鼠卵巢及子宫结构完整；与对照组相比，模型组大鼠子宫腔缩小，卵巢多囊化严重，血清T、FSH、LH水平均显著提高，TGF-β1、SMAD2、SMAD3mRNA及蛋白表达水平显著升高（P＜0.05）；与模型组比较，阳性药物二甲双胍及六味地黄丸处理后，大鼠卵巢多囊化明显缓解，子宫腔扩张，血清T、FSH、LH水平均显著降低，TGF-β1、SMAD2、SMAD3mRNA及蛋白表达水平显著降低（P＜0.05），且两组以上指标表达无显著差异；与六味地黄丸组相比，六味地黄丸＋SRI-011381组大鼠卵巢多囊化加重，血清T、FSH、LH水平均显著升高，TGF-β1、SMAD2、SMAD3mRNA及蛋白表达水平显著升高（P＜0.05）。结论 六味地黄丸可有效抑制TGF-β1、SMAD2、SMAD3mRNA及蛋白的表达，缓解卵巢多囊化、子宫腔扩张，调节内分泌代谢，从而改善PCOS病情。

Objective To establish a rat model of polycystic ovary syndrome (PCOS), to explore whether Liuwei Dihuang Pill (LDP) affects endocrine metabolism in PCOS rats by regulating TGF-β1/SMAD signaling pathway.Methods Sixty SD female rats were randomly divided into control group, model group, metformin (positive drug, 50 mg·kg^-1) group, LDP (50 mg·kg^-1) group, LDP (50 mg·kg^-1)+SRI-011381 (TGF-β1/SMAD pathway activator, 100 ng·kg^-1) group. Except the control group, the other groups were ig with 1 mg·kg^-1 letrozole (dissolved in 0.5% sodium carboxymethyl cellulose) and fed high-fat diet every day for 15 days to establish PCOS rat model. The control group was ig 0.5% sodium carboxymethyl cellulose daily and fed ordinary diet. After modeling, each group was ig given the drug, and the control group and model group were given the same volume 0.9% sodium chloride solution intragaically, respectively, for consecutive 10 days. Hematoxylin-eosin (HE) staining method was used to detect the pathological changes of the ovaries and uterus of rats in each group, enzyme-linked immunoassay kit (ELISA) method was used to detect levels of serum testosterone (T), luteinizing hormone (LH), and follicle stimulating hormone (FSH) of rats in each group, fluorescence quantitative PCR (qRT-PCR) was used to detect the expression levels of TGF-β1, SMAD2, and SMAD3 mRNA in the ovarian tissues of each group, Western blotting was used to detect the expression levels of TGF-β1, SMAD2, and SMAD3 proteins in ovarian tissues of each group.Results The structures of the ovary and uterus in the control group were complete. Compared with the control group, the uterine cavity of the model group was reduced, the ovarian polycystication was serious, the levels of serum T, FSH, and LH were significantly higher, and the expression levels of TGF-β1, SMAD2, SMAD3 mRNA and protein were significantly higher (P <0.05). Compared with model group, after treatment with the metformin and LDP, the rat ovarian polycystication was significantly relieved, the uterine cavity was dilated, the levels of serum T, FSH, and LH were significantly reduced, and the expression levels of TGF-β1, SMAD2, SMAD3 mRNA and protein were significantly reduced (P <0.05), and there was no significant difference in the expression of indicators between the two groups. Compared with the medication group, the polycystication of the ovaries in the rats in the medication + SRI-011381 group increased significantly, the levels of serum T, FSH, and LH were significantly increased, and the expression levels of TGF-β1, SMAD2, SMAD3 mRNA and protein were significantly increased (P <0.05).Conclusion LDP can effectively inhibit the expression of TGF-β1, SMAD2, SMAD3 mRNA and protein, relieve ovarian polycystication and uterine cavity dilatation, regulate endocrine metabolism, and improve the condition of PCOS.

R285.5

广东省中医药局科研项目(20201032)