基于SCF/c-kit--PI3K--Bcl-2通路探讨补肾生精药对“菟丝子-枸杞子”调控生精细胞增殖、凋亡的作用机制

Male infertility has become a global problem in medicine and sociology, the abnormal spermatogenesis is one of the major causes of its occurrence which needs for safe and effective treatment. The previous study in our research group confirmed that the "Zuogui Pills" can promote the proliferation of spermatogenic cells and can also inhibit their apoptosis. "Dodder、medlar" is "the core herbs in "Zuogui Pills" which are also the key medicines for the deficient or weak sperms in TCM, so whether it has a similar effect with the Zuogui pill? As we know the SCF/c-kit--PI3K--Bcl-2 pathway is an important pathway for regulating the proliferation and apoptosis of spermatogenic cells in vivo. Does the herbs’ combination of "dodder-medlar" have a regulatory effect on it? Based on this study, this research takes the SCF/c-kit--PI3K--Bcl-2 signaling pathway as a starting point via the construction of dyszoospermia rat model in vivo and isolated spermatogenic cell injury model in vitro. we focus on the sperm quality, reproductive endocrinology, testicular morphology and ultrastructural changes, spermatogenic cell proliferation, apoptosis and the expression of protein in SCF/c-kit--PI3K--Bcl-2 pathway. Through the above researchs we explore the effects of "dodder-medlar" on spermatogenic functio and its possible molecular mechanism in order to provide a scientific basis for the theroy kidney-nourishing and essence-generating therapy in treating oligoasthenozoospermia.mechanism in order to provide a scientific basis for the theroy of Bushentianjing in treating oligoasthenozoospermia.

男性不育症已成为全球性的医学和社会学问题，生精功能异常是导致其发生的主要病因之一，亟需安全有效的治疗手段。课题组前期研究证实“左归丸”具有促进生精细胞增殖、抑制其凋亡的作用。“菟丝子-枸杞子”是“左归丸”中的核心药对，也是中医临床上治疗少弱精子症的关键药物。其是否具有与左归丸相似的效应？SCF/c-kit--PI3K--Bcl-2通路是体内调控生精细胞增殖与凋亡的重要通路，“菟丝子-枸杞子”药对对其是否具有调控效应？基于此，本研究以SCF/c-kit--PI3K--Bcl-2信号通路为切入点，通过构建在体生精障碍大鼠模型、离体生精细胞损伤模型，并观察大鼠精液质量、生殖内分泌、睾丸形态学与超微结构变化、生精细胞增殖、凋亡及SCF/c-kit--PI3K--Bcl-2通路蛋白的表达情况，探讨“菟丝子-枸杞子”药对对生精功能的影响及其可能的分子机制，以期为补肾生精法治疗少弱精子症提供一定的科学依

随着环境、食品、心理等因素的影响以及生活方式的改变，男性不育的发病率逐年增加，世界范围内，不孕不育的发病率已提升到15%-18%。生精功能异常是导致其发生的主要病因之一，亟需安全有效的治疗手段。课题组前期研究证实“左归丸”具有促进生精细胞增殖、抑制其凋亡的作用，“菟丝子-枸杞子”是“左归丸”中的核心药对，也是中医临床上治疗少弱精子症的关键药物，其是否具有与左归丸相似的效应不得而知，且作用机制也尚不明确。本课题以“菟丝子-枸杞子”药对干预生精功能障碍模型大鼠的体内外实验研究为主要内容，来探讨“菟丝子-枸杞子”药对改善生精功能的潜在作用机制。实验结果发现，经“菟丝子-枸杞子”药对干预后，GTW＋SCFL组的SCF、c-kit、PI3K、p-Akt、Bcl-2的蛋白及mRNA水平均较GTW组有明显的增加，BAD和Bax蛋白及mRNA的表达水平有明显的下调（P＜0.05），且在超微结构上，GTW＋SCFL组睾丸组织的初级精母细胞中可见大量线粒体，次级精母细胞中可见精子颗粒，精子细胞数量增加，且结构损伤得到改善。说明“菟丝子-枸杞子”药对可对SCF/c-kit-PI3K-Bcl-2信号通路起到一定的调控作用，增加生精细胞增殖，抑制细胞线粒体与生精细胞的凋亡，改善生精功能障碍。本研究以“肾主生殖”中医理论为切入点，以“菟丝子-枸杞子”这一治疗男性不育症传统名方的核心药对为研究对象，从分子机制阐明了菟丝子-枸杞子”药对可以通过调控SCF/c-kit-PI3K-Bcl-2信号通路促进生精细胞发育、抑制其凋亡、进而提高精液质量。本研究的开展为“肾主生殖”这一中医理论提供了实验基础，对于促进中药治疗男性不育症的临床应用具有重要意义。