基于ROS-线粒体途径的补肾活血法调控男性不育症自噬与凋亡机制研究

Previous studies demonstrated that traditional Chinese medicine Yi Jing Fang can be effective in the treatment of oligoasthenozoospermia, and be able to reduce the level of reactive oxygen species in seminal plasma, improve sperm mitochondrial membrane potential, inhibition of early sperm apoptosis. Autophagy and apoptosis combined the latest developments, and put forward the hypothesis 1 mitochondrial autophagy this specific route of elimination, regulation may be subtle green environment ROS level and spermatogenic cell apoptosis, autophagy balance between self-ties, hypotheses 2 traditional Chinese medicine from oxidative stress, mitochondrial function protection and stability, regulating autophagy and apoptosis balance, improve student subtle environment. In front less weak azoospermia treatment of patients with idiopathic sperm samples, CP-induced oxidative stress in mice infertile model, Leydig cell damage induced in vitro ACR model object to ROS and mitochondrial function as the center and explore LC3, Beclin-1 , PI3K / AKT / mTOR autophagy and other factors, and Caspase-3, Bcl-2 and Bax apoptosis and other factors related to spermatogenic cell autophagy and apoptosis regulation, in-depth interpretation of the treatment of kidney and promoting blood circulation less weak azoospermia the molecular mechanism for the rich scientific connotation of Chinese medicine theory, and constantly improve the efficacy of service.

前期研究证实，补肾活血中药益精方能够有效治疗少弱精子症，并能够降低精浆活性氧水平，提高精子线粒体膜电位，抑制精子早期凋亡。结合自噬与凋亡最新进展，提出假说1线粒体及其自噬这一特异性的清除途径，可能是调控生精微环境ROS 水平以及生精细胞发生凋亡、自噬之间平衡的纽带，假说2补肾活血中药可从抗氧化应激，保护线粒体功能稳定，调节自噬与凋亡平衡，改善生精微环境。以特发性少弱精症患者治疗前后精子标本、CP诱导的小鼠氧化应激不育模型、体外ACR诱导Leydig细胞损伤模型为对象，以ROS与线粒体功能为中心，探讨LC3、Beclin-1、PI3K/AKT/mTOR等自噬因子，与Caspase-3, Bcl-2和Bax等凋亡因子，对生精相关细胞自噬与凋亡的调控作用，深入阐释补肾活血方药治疗少弱精症的分子机制，为丰富中医药理论科学内涵、不断提高疗效服务。

男性不育症是男科临床的常见病和多发病，中国男性近30年来，精子密度、精子非前向运动率(C%)呈显著下降趋势。多种原因如吸烟、酗酒、毒物暴露、电离辐射；精索静脉曲张、隐睾、肥胖等都会造成雄性（男性）生殖微环境相关细胞的氧化应激损伤，从而造成少、弱、畸精症等，即睾丸氧化应激损伤是男性不育的重要病理因素和共同特征，但其中具体涉及到的关键因子及信号转达途径尚不清晰。本项目在前期研究的基础上，通过体内-体外-临床三个层次，纵向观察，结合运用蛋白质生物化学、细胞生物学等方法和技术，从活性氧-线粒体介导生精细胞自噬与凋亡的相互作用，深入探讨益精方改善精子质量的作用机制。结果显示，益精方可以改善氧化应激导致的睾丸生精小管结构损伤、生精环境破坏、自噬-凋亡失平衡，通过PI3K/AKT/mTOR信号途径，改善ROS与线粒体功能异常，起到凋亡和自噬蛋白分子间的协调平衡作用，进而为正常的精子发生及成熟等提供保障，最终提高精子活力，改善精子畸形率，增加精子密度，达到治愈男性不育的目的。此研究为进一步提高临床疗效，奠定了理论基础。