基于肠道菌群-炎性反应-胰岛素抵抗途径的中药干预多囊卵巢综合征的分子机制研究

Insulin resistance（IR）is a hinge which connects reproduction abnormality and metabolic abnormality in polycystic ovary syndrome(PCOS). Gut microbiota imbalance can cause the release of inflammatory cytokines and effect the serine phosphorylation of insulin receptor substrate, also associated with hyperandrogenism. Banxiaxiexin decoction can regulate gut microbiota and ameliorate the state of IR. Our previous research has found that Banxiaxiexin decoction can ameliorate the state of IR in treating the patients with PCOS. Therefore, we propose that Banxiaxiexin decoction plays a role in the treatment of PCOS by regulating the gut microbiota, improving the inflammatory state and ameliorating insulin sensitivity. Taking Banxiaxiexin decoction as intervention，this research adopts metagenome, bioinformatics, RT-PCR and Western blotting to analysis the effect of Banxiaxiexin decoction in treating PCOS IR mouse model through the characteristic flora spectrum, specific inflammatory cytokines and insulin signaling pathway so as to interpret the objectivity of the hypothesis. It provides scientific basis for classical prescription in the clinical application and promotion as well as experience and inspires a novel thinking for the research and treatment of PCOS. It reveals the scientific connotation of "chong channel in favor of Yangming" from another side and achieves a new level in treating PCOS with traditional Chinese medicine from theory to practice. Similar articles have not been reported in domestic and foreign countries.

胰岛素抵抗（IR）是连接多囊卵巢综合征（PCOS）生殖异常和代谢障碍的重要枢纽。肠道菌群失调能引起炎性因子释放，影响胰岛素受体底物丝氨酸磷酸化导致IR，还与高雄激素血症有关。半夏泻心汤具有调整胃肠道菌群，改善IR作用。我们的前期研究也发现，半夏泻心汤加减能改善IR，治疗PCOS。因此提出“半夏泻心汤通过调整肠道菌群、改善炎症状态，提高胰岛素敏感性而发挥治疗PCOS 作用”假说。本研究以半夏泻心汤加味为干预手段，采用元基因组学、生物信息学、RT-PCR、Western blotting等技术探讨中药对PCOS IR小鼠模型肠道菌群、特异性炎症因子、胰岛素信号通路相关因子等的影响，以验证该假说的客观性，为经方的临床应用和推广提供科学依据，为PCOS的研究和治疗提供借鉴和开辟新的思路；从一个侧面揭示“冲脉隶于阳明” 的科学内涵，使中医药治疗PCOS从理论到实践达到新的水平。国内外尚未见同类报道。

本研究从肠道菌群-炎性反应-胰岛素抵抗途径探讨中药治疗PCOS的分子机制，结果显示，与模型组相比，中药组①大鼠体重降低，卵巢脏器系数升高。②组织病理学检查，镜下可见各级卵泡及多个黄体，卵泡颗粒细胞呈多层，偶见囊状扩张卵泡。③血清T水平有下降趋势； FBG和FINS有下降趋势，HOMA-IR水平下降；LPS有下降趋势，TNFα、IL-10降低。④肠道菌群检测，非共有OUT减少，实际观测的OTU 数目明显下降，群落丰富度极显著下降，群落多样性显著下降。微生物群落间的物种多样性有差异。在门水平，疣微菌门丰富度有上升趋势，拟杆菌门有下降趋势，放线菌门下降，变形菌门升高。在属水平，狭义梭菌属明显下降。差异性物种主要为葡萄球菌科(Staphylococcaceae) 、葡萄球菌属(Staphylococcus)、 γ-变形菌纲(Gammaproteobacteria)、变形菌门(Proteobacteria)、肠杆菌目 Enterobacteriales 、拟杆菌属(Bacteroides)、拟杆菌科(Bacteroidaceae) 、Monoglobus菌属、Monoglobaceae菌科、Dubosiella菌属。体重、E2、FSH、LH、FINS、T、LH/FSH、FINS、HOMA-IR、Glu、LPS等均对细菌群落组成有显著影响。⑤在卵巢、脂肪和肝脏等组织， IL-10基因和蛋白表达均升高； TNF-α蛋白表达降低。JNK、IRS1、IRS2、STAT3等蛋白表达均降低。结论：半夏泻心汤加味通过影响肠道菌群的群落、菌群多样性和门、属等结构，减轻炎性反应，影响胰岛素信号通路的信号传导，发挥治疗PCOS的作用。