基于人精子线粒体膜蛋白PHB表达探索石萆汤治疗肾虚夹湿型弱精子症的分子机制

Asthenospermia is one of the common causes of male infertility, accounting for about 19%. Previous studies have confirmed that Shibi decoction asthenospermia have a significant effect, can improve the structure and function of the state of sperm mitochondria. Sperm mitochondrial membrane protein Prohibitin (PHB) is highly conserved, mainly in the inner mitochondrial membrane expression, involved in the formation of mitochondria, associated with respiratory chain, mitochondrial division assisting Guinea space. Thus, we speculate that male infertility caused by abnormal sperm motility, which may be related to abnormal expression of PHB, which may be Shibi decoction treatment efficacy asthenospermia base and target. In this study, before and after Syndrome of Kidney Deficiency Complicated with Dampness asthenospermia patients as objects, according to the WHO criteria to evaluate sperm motility using MitoTracker ® probes staining, flow cytometry analysis combined with sperm mitochondrial function, and by immunofluorescence Western blot Western blot analysis of sperm mitochondrial membrane protein cellular localization and expression of PHB. PHB protein-based, clear Shibi decoction treatment of Syndrome of Kidney Deficiency Complicated with Dampness asthenospermia molecular mechanisms.

弱精子症是导致男性不育的常见原因之一，约占19%。既往研究证实我院男科名老中医经验方石萆汤治疗弱精子症有显著疗效，可改善精子线粒体的结构和功能状态。精子线粒体膜蛋白Prohibitin(PHB)高度保守，主要在线粒体内膜表达，参与线粒体嵴的形成，与呼吸链相关，协助划分线粒体内亚空间。由此我们推测因精子活力异常所致的男性不育，其发生可能与PHB表达异常有关，可能是石萆汤治疗弱精子症的疗效基础与靶点。本研究以石萆汤治疗前后的肾虚夹湿型弱精子症患者精子为对象，根据WHO标准评价精子活力，利用MitoTracker®探针染色，结合流式细胞术检测分析精子线粒体功能，通过免疫荧光与Western blot免疫印迹技术分析精子线粒体膜蛋白PHB的细胞定位与表达变化。基于人精子线粒体膜蛋白PHB表达，明确石萆汤治疗肾虚夹湿型弱精子症的分子机制。

弱精子症是导致男性不育的常见原因之一，约占19%。既往研究证实福建中医药大学附属人民医院协定方石萆汤治疗弱精子症有显著疗效。精子线粒体膜蛋白Prohibitin(PHB)高度保守，主要在线粒体内膜表达，参与线粒体嵴的形成，与呼吸链相关。因精子活力异常所致的男性不育，其发生机制与线粒体膜蛋白PHB表达异常有关。中医认为弱精子症基本病机与肾虚、湿阻等有关，石萆汤针对此病机具有利湿通窍、补肾填精的功效。本研究基于前期研究，入选以石萆汤治疗的肾虚夹湿型弱精子症60例患者（T组）、正常体检者30例（N组）、检测为弱精子暂未采取治疗措施者30例（A组）的精子为对象，根据WHO（第四版）标准评价精子活力，采用Western blot检测患者精子线粒体膜蛋白PHB及JC-1流式细胞仪检测线粒体膜电位；采用荧光探针MitoTracker，激光扫描共聚焦显微镜(LSCM) 观察精子细胞的荧光强度，分析精子细胞线粒体膜蛋白PHB的阳性细胞比例；用透射电镜观察精子细胞线粒体超微结构的变化。通过对以上结果的分析，精子的总活力以及前向运动的精子的比率与线粒体膜蛋白PHB、线粒体膜电位表达呈正相关，与中医症状评分呈负相关线性关系，石萆汤通过上调精子线粒体膜蛋白PHB表达，提高线粒体膜电位，改变精子细胞线粒体超微结构，激发精子的线粒体功能，从而治疗弱精子症，同时石萆汤有改善肾虚夹湿证候的临床症状。因此，通过以上研究结果，可以阐明基于人精子线粒体膜蛋白PHB表达探索石萆汤治疗肾虚夹湿型弱精子症的分子机制，为石萆汤治疗肾虚夹湿型弱精子症的中西医结合基础研究的深化奠定阶段性的基础。