补肾活血方对卵巢早衰大鼠颗粒细胞端粒酶活性的影响

With the modification of the modern society lifestyle, the incidence of decreasing ovarian store (DOS) and premature ovarian failure (POF) was significantly increased. DOS can induce POF. Timely chinese herbs intervention can effectively prevent or delay the occurrence of POF. Recently, it was discovered that the ovarian granulosa cell has the stem cell proliferative potential, its expression of telomerase activity may be a more direct indication of ovarian reserve. .The project intends to observe the preventive and curative effect of BHF on DOS and POF in the ovariectomized rats, to clarify its action on the ovarian granulosa cell's function, to explore its mechanism for telomerase activity by the three MAPK signaling pathway p38/ERK1/2/JNK and break the clinic therapeutic limitation for DOS and POF. The Department of Gynecology of Shanghai Longhua Hospital has many years of clinical experience in the field of the prevention and treatment of DOS. It develops the diagnosis and treatment standard characterized by "invigorating kidney and promoting blood flow". Our preliminary experiments have showed that the model of DOS was established and BHF was effective. .The project is to set forth the scientific implications of the theory that ''The kidney governs reproduction and stores essence" with the telomerase activity of granulosa cell,to build relationship between molecular mechanism of aging and the theory of "kidney essence". Thus, it can enrich and expand our knowledge and research ideas of the Traditional Chinese Medicine (TCM) theory "The kidney stores essence" by the perspective of procreation axis named "Kidney-TianGui-ChongRen meridians-womb".

现代人生活方式的改变，使卵巢储备功能下降（DOS）和卵巢早衰（POF）的发病率逐年升高，DOS很可能进一步发展为POF，及时进行中药干预可有效的防止或延缓POF的发生。卵巢颗粒细胞具备干细胞增殖潜能，其表达的端粒酶活性可以更加直接的反应卵巢储备功能。.本项目拟观察补肾活血方防治去势大鼠卵巢DOS和POF疗效，明确其对卵巢颗粒细胞功能的作用，从MAPK三条信号通路p38/ERK1/2/JNK探索对端粒酶活性的作用机制，突破临床对DOS和POF的治疗局限。龙华医院妇科防治DOS的临床经验丰富，形成了以"补肾活血"为特色的临床诊疗规范。前期实验已证实DOS模型成立，补肾活血方疗效确定。.本项目从颗粒细胞端粒酶活性阐述"肾主生殖"与"肾藏精"理论部分科学内涵，为联系衰老分子机制与"肾精"理论之间搭起桥梁。从"肾－天癸－冲任－胞宫"生殖轴角度丰富提高中医"肾藏精"理论的认识和研究思路。

卵巢早衰（POF）是卵巢功能不全的典型表现，其可诱发卵巢储备功能下降（DOS）进而导致不孕症发生。因此，探索POF的发病机制并及时进行中药干预可有效的防止或延缓其发生发展。本课题主要探索了雷公藤和环磷酰胺所致的卵巢化疗损伤的发病机制，并探讨了中药“补肾活血方”对POF模型鼠的治疗作用及其作用靶点。通过POF模型的建立、卵巢组织病理学分析、生物化学与分子生物学检测手段的运用，我们阐明了如下机理：在POF发生机制方面，我们不仅阐明雷公藤多苷通过激活卵巢颗粒细胞Stk11-p53通路，刺激p53激活下游基因p21的转录，诱发小鼠POF的作用机制；而且，我们还阐明了，环磷酰胺显著促进小鼠卵巢颗粒细胞内源性lncRNA-Meg3的表达，维持p53的活性，进一步活化衰老相关基因p66SHC和p16的表达，最终诱发细胞衰老和凋亡，从而导致小鼠POF的发生机制。在治疗方面，我们阐明了“补肾活血方”通过活化MAPK三条信号通路p38/ERK1/2/JNK，诱发端粒酶活性，降低Bax及Caspase-3蛋白的表达，延缓雷公藤对大鼠卵巢颗粒细胞损伤作用，从而实现对卵巢早衰的治疗作用。上述研究结果，从卵巢颗粒细胞活性角度阐述“肾主生殖”与“肾藏精”理论部分科学内涵，为联系衰老分子机制与“肾精”理论之间搭起桥梁。从“肾－天癸－冲任－胞宫”生殖轴角度丰富提高中医“肾藏精”理论的认识和研究思路。为“补肾活血方”治疗POF的临床运用提供了实验数据的支持。