二甲雙胍治療多囊卵巢綜合征機制研究

蘆晨宇+秦培鋼++杜函妤++鄔海霞++古麗亞爾·艾合買提+艾麗霞++徐琦

[摘要] 目的 探討二甲雙胍在治療多囊卵巢綜合征（PCOS）中對Treg細胞和Th17細胞的作用及其機制。 方法 雌性Wistar大鼠45只，來曲唑灌胃建立PCOS大鼠模型，隨機抽取5只確立成模后，按完全隨機分組原則分為二甲雙胍組[300 mg/（kg·d）]、達英組[180 mg/（kg·d）]、二甲雙胍[300 mg/（kg·d）]與達英[180 mg（kg·d）]聯(lián)合治療組及模型組，每組10只；另選取10只大鼠作為對照組。成模后，各治療組采用相應藥物灌胃治療，模型組給予等量生理鹽水，均連續(xù)處理28 d，觀察各組大鼠每周體質(zhì)量變化。蘇木精-伊紅（HE）染色法觀察卵巢組織學變化，流式細胞術檢測脾臟中Treg細胞和Th17細胞數(shù)量及表型的改變，ELISA法檢測血清中IL-6和TGF-β水平，RT-PCR法測定卵巢組織轉(zhuǎn)錄因子Foxp3、ROR-γt的表達水平。 結果 模型組大鼠體質(zhì)量、脾臟中Treg細胞、Th17細胞數(shù)量以及Treg/Th17比例、血清中IL-6及TGF-β水平以及卵巢組織中轉(zhuǎn)錄因子Foxp3的相對表達量顯著高于對照組（P < 0.05）；二甲雙胍組、達英組與聯(lián)合治療組體質(zhì)量與模型組相比顯著降低（P < 0.05），與對照組比較差異無統(tǒng)計學意義（P > 0.05）；二甲雙胍組、達英組及聯(lián)合用藥組囊性擴張卵泡較模型組明顯減少；二甲雙胍組脾臟Treg細胞、Th17細胞、血清IL-6水平及卵巢組織中轉(zhuǎn)錄因子ROR-γt的表達水平較模型組顯著升高（P < 0.05），達英組及聯(lián)合用藥組脾臟Th17細胞、血清IL-6水平與模型組相比顯著降低（P < 0.05），而卵巢組織中轉(zhuǎn)錄因子Foxp3的表達水平較模型組顯著升高（P < 0.05）。 結論 二甲雙胍治療PCOS的作用機制可能是通過影響血清細胞因子IL-6表達水平，調(diào)節(jié)Treg、Th17細胞的數(shù)量及比例，進而改善卵巢組織微環(huán)境。

[關鍵詞] 多囊卵巢綜合征；Treg；Th17；二甲雙胍

[中圖分類號] R392.1 [文獻標識碼] A [文章編號] 1673-7210（2017）11（c）-0017-05

Study on the mechanism of Metformin in the treatment of polycystic ovary syndrome

LU Chenyu QIN Peigang DU Hanyu WU Haixia Aihemaiti·Guliyaer AI Lixia XU Qi

Basic Medical College， Xinjiang Medical University， Xinjiang Uygur Autonomous Region， Urumqi 830011， China

[Abstract] Objective To investigate the therapeutic mechanism of Metformin on Treg cells and Th17 cells in polycystic ovary syndrome （PCOS）. Methods 45 female Wister rats were gavaged by Letrozole in order to develop PCOS， and randomly chosen 5 rats to confirm the establishment of model. Then rats were divided into Metformin group [300 mg/（kg·d）]， Diane group [180 mg/（kg·d）]， Metformin [300 mg/（kg·d）] combined with Diane [180mg/（kg·d）] group and model group， randomly， each group had 10 rats. The other 10 female rats were arranged as control group. PCOS rats were given corresponding drugs， while control group were administrated with physiological saline， the treatment period of all groups were 28 days. The weight changes of rats weekly were observed. Ovarian histological changes were examined by Yihong-hematoxylin staining （HE）. Flow cytometry was applied to delineate the phenotype of Treg cells and Th17 cells in spleen. ELISA was used to measure IL-6 and TGF-β in serum. The expression levels of Foxp3 and ROR-γt were measured by RT-PCR. Results The weight， the Treg and Th17 in spleen， Treg/Th17 ratio， the levels of IL-6 and TGF-β in serum， the expression of Foxp3 of PCOS model group were higher than control， with statistically significant difference （P < 0.05）. The weight of Metformin group， Diane group and combined group were similar to control group （P > 0.05）， but significantly lower than model group （P < 0.05）. The cystic dilated follicle in Metformin group， Diane group and combined group decreased significantly compared with the model group. Treg cells and Th17 cells in spleen， IL-6 and transcription factor ROR-γt in serum of the Metformin group were significantly higher than the model group （P < 0.05）. Th17 cells in spleen and IL-6 in serum of Diane group and the combined group were significantly lower than the model group （P < 0.05）， but the expressions of transcription factor Foxp3 in ovarian tissue were significantly higher than the model group （P < 0.05）. Conclusion Metformin treatment of PCOS may affect the number and proportion of Treg and Th17 cells by affecting the expression level of the serum IL-6， which can improve the microenvironment of ovarian tissue.endprint

[Key words] PCOS； Treg； Th17； Metformin

多囊卵巢綜合征（polycystic ovary syndrome，PCOS）是一種常見于青春期及育齡婦女的內(nèi)分泌紊亂性疾病，其主要表現(xiàn)為月經(jīng)失調(diào)、持續(xù)性無排卵及不孕，常伴有肥胖、高胰島素血癥和胰島素抵抗等癥狀[1]。PCOS在我國的患病率約為6.5%，且有逐年升高趨勢[2]。研究表明，慢性炎癥能介導胰島素抵抗（IR）的發(fā)生發(fā)展，是PCOS可能的發(fā)病因素之一[3]。……