经APN信号通路探讨补肾化痰法调控肥胖型PCOS卵泡发育机制研究

This research focused on kidney deficiency and dampness-phlegm, the key pathogenesis of obese PCOS. In this study, APN/p38MAPK /PPAR signal transduction pathways ( APN signal transduction pathways) as the main line, which not only can reflect adipose tissue function , but also is closely related with folliculogenesis. We Put forward the hypothesis that the principle of tonifying kidney and reducing phlegm may promote the obese PCOS folliculogenesis through regulating the APN p38MAPK/PARR signaling pathhway . Ob/ob mice molding obese PCOS , GC in vitro culture techniques, western hybridization and RT-PCR technology are employed in our research. From the three levels of the overall ,cellular and molecular, the mechanism of tonifying kidney and reducing phlegm methods about regulation GC proliferation and sex hormone synthesis will be explored. By observing the body glucolipid metabolism, insulin resistance effect of the change, the mechanism of tonifying kidney and reducing phlegm method regulation of the obese PCOS folliculogenesis will be revealed. Providing a scientific basis for internal relation of further interpretation fueling PCOS folliculogenesis disorders associated with reproductive endocrine disorder, IR and sugar, lipid metabolism disorder; scientific basis for TCM treatment of obese PCOS, the scientific connotation of tonifying kidney and reducing phlegm is enriched.

针对肥胖型PCOS“肾虚痰湿内蕴”的关键病机，本研究以能体现脂肪组织功能与卵泡发育密切相关的APN/ p38MAPK/PPAR信号转导通路（APN信号通路）为主线，提出“补肾化痰法可能通过调控APN信号通路进而促进肥胖型PCOS卵泡发育”之假说。采用SD大鼠制备肥胖型PCOS模型、GC体外培养技术、Western杂交、RT-PCR等关键技术，从整体、细胞及分子水平三个层次探索补肾化痰法调控卵泡GC增殖、生殖性激素合成的机制，并通过观察、胰岛抵抗效应、糖脂代谢的变化，揭示补肾化痰法经APN信号通路调控肥胖型PCOS卵泡发育的机制，为深入阐释肥胖型PCOS卵泡发育障碍及相关生殖性激素紊乱、胰岛素抵抗及糖脂代谢与APN信号通路的内在联系提供科学依据，又为中医治疗肥胖型PCOS提供科学依据，并进一步丰富中医补肾化痰治法的科学内涵。

1本课题主要针对肥胖型PCOS“肾虚痰湿内蕴”的关键病机，选取SD雌性大鼠，用来曲唑联合高脂灌服制备肥胖型PCOS模型。采用放射免疫法、ELISA法、Western blotting及Realtime PCR分别检测模型大鼠血清性激素、卵巢颗粒细胞（GC）中APN信号途径信号分子的表达以及与GC增殖分化相关的信号分子；结果发现肥胖型PCOS模型大鼠出现排卵障碍，生殖内分泌紊乱的表现，并伴随全身的糖脂代谢异常及胰岛素抵抗（IR）。以代表补肾化痰法的经典复方治疗后，肥胖型PCOS模型雌鼠排卵障碍、生殖内分泌、糖脂代谢异常及IR明显改善，推测补肾化痰法可能经APN信号通路改善肥胖型PCOS卵泡发育的机制，为深入阐释肥胖型PCOS卵泡发育障碍及相关引起的糖脂代谢异常、生殖内分泌紊乱及IR与APN信号通路的内在联系提供依据，为中医临床运用传统补肾化痰这一治法深入研究治疗肥胖型PCOS奠定了科学的实验基础。