目的 建立多囊卵巢綜合征（PCOS）大鼠模型，探究六味地黃丸是否通過(guò)轉(zhuǎn)化生長(zhǎng)因子β1（TGF-β1）/SMAD信號(hào)通路對(duì)PCOS大鼠內(nèi)分泌代謝產(chǎn)生影響。方法 將60只SD雌性大鼠隨機(jī)分為對(duì)照組、模型組、二甲雙胍（陽(yáng)性藥，50mg·kg^-1）組、六味地黃丸（50mg·kg^-1）組、六味地黃丸（50mg·kg^-1）＋SRI-011381（TGF-β1/SMAD通路激活劑，100ng·kg^-1）組，除對(duì)照組外，其余各組每天ig1mg·kg^-1來(lái)曲唑（溶于0.5%羧甲基纖維素鈉），并喂食高脂飼料，連續(xù)15d，構(gòu)建PCOS大鼠模型；對(duì)照組每日ig等體積0.5%羧甲基纖維素鈉，并喂食普通飼料。造模完成后，各組ig給藥，對(duì)照組、模型組分別ig等體積0.9%氯化鈉溶液，連續(xù)10d。蘇木精-伊紅（HE）染色法檢測(cè)各組大鼠卵巢、子宮的病理變化情況；采用酶聯(lián)免疫試劑盒（ELISA）法檢測(cè)各組大鼠血清睪酮（T）、黃體生成素（LH）、卵泡刺激素（FSH）水平；采用實(shí)時(shí)熒光定量PCR（qRT-PCR）法檢測(cè)各組卵巢組織中TGF-β1、SMAD2、SMAD3mRNA的表達(dá)水平，采用Western blotting法檢測(cè)各組卵巢組織中TGF-β1、SMAD2、SMAD3蛋白表達(dá)水平。結(jié)果 對(duì)照組大鼠卵巢及子宮結(jié)構(gòu)完整；與對(duì)照組相比，模型組大鼠子宮腔縮小，卵巢多囊化嚴(yán)重，血清T、FSH、LH水平均顯著提高，TGF-β1、SMAD2、SMAD3mRNA及蛋白表達(dá)水平顯著升高（P＜0.05）；與模型組比較，陽(yáng)性藥物二甲雙胍及六味地黃丸處理后，大鼠卵巢多囊化明顯緩解，子宮腔擴(kuò)張，血清T、FSH、LH水平均顯著降低，TGF-β1、SMAD2、SMAD3mRNA及蛋白表達(dá)水平顯著降低（P＜0.05），且兩組以上指標(biāo)表達(dá)無(wú)顯著差異；與六味地黃丸組相比，六味地黃丸＋SRI-011381組大鼠卵巢多囊化加重，血清T、FSH、LH水平均顯著升高，TGF-β1、SMAD2、SMAD3mRNA及蛋白表達(dá)水平顯著升高（P＜0.05）。結(jié)論 六味地黃丸可有效抑制TGF-β1、SMAD2、SMAD3mRNA及蛋白的表達(dá)，緩解卵巢多囊化、子宮腔擴(kuò)張，調(diào)節(jié)內(nèi)分泌代謝，從而改善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

廣東省中醫(yī)藥局科研項(xiàng)目(20201032)