从SCF/c-kit系统研究调肝中药干预VC性不育的机理

Varicocle (VC) is the first reason causing male infertility, and our previous researches and clinical trials showed that liver-regulating herb had a good curative effect on VC. We found that stem cell factor (SCF) and its receptor c-kit changed significantly in experimental VC rats treated with liver-regulating herb when compared to control rats. According to literatures, we deduced that the herb may play its curative role through interfering with SCF/c-kit system. Therefore, this program will explore the effect of liver-regulating herb on reproductive ability of infertile male rats with experiment VC in vivo, detect the expression of the receptor of FSH on sertoli cells and c-kit on spermatogenic cells at mRNA and protein levels, and the protein expression of SCF, observe the con-location of SCF/c-kit on testis tissue, and analyze the relativity of curative effect of the herb on male infertility caused by VC and SCF/c-kit system. In vitro, we will study the effect of the liver-regulating herb on SCF/c-kit under the three different cell cultures, including sertoli cells and spermatogenic cells individually, and the both cells, and investigate the effect of SCF/c-kit on the herb improving the spermatogenesis function. So then, we will supply enough evidence to prove the curative effect of the liver-regulating herb on male infertility with VC, clarify this effect is mediated by SCF/c-kit system and SCF/c-kit is the target of the liver-regulating herb intervening in male infertility with VC.

精索静脉曲张（VC）是男性不育的第一大原因，我们前期的理论探讨、临床研究发现调肝中药具有较好疗效，动物实验发现干细胞因子（SCF）及其受体c-kit在调肝中药治疗时发生了显著变化，结合文献，我们认为调肝中药可能是通过干预SCF/c-kit系统发挥治疗作用的。本研究拟在体内观察调肝中药对VC不育大鼠生殖能力的作用，检测支持细胞FSH受体和生精细胞c-kit mRNA和蛋白表达、SCF蛋白表达，观察SCF/c-kit在睾丸组织的共定位表达，分析调肝中药对VC不育的治疗作用及其与SCF/c-kit系统的相关性；在体外研究调肝中药对体外培养的支持细胞、生精细胞及支持细胞/生精细胞共存条件下SCF/c-kit系统的影响，探讨SCF/c-kit系统在调肝中药改善生精功能中的作用。从而验证调肝中药是治疗VC不育的有效方法，阐明VC导致不育是通过SCF/c-kit介导的，是调肝中药干预VC不育的靶点。

精索静脉曲张（VC）是男性不育的第一大原因，但其导致男性不育的具体机制尚不明确。我们前期的理论探讨、临床研究发现调肝中药具有较好疗效，动物实验发现干细胞因子（SCF）及其受体c-kit在调肝中药治疗时发生了显著变化，结合文献，我们认为调肝中药可能是通过干预SCF/c-kit系统发挥治疗作用的。本研究在体内研究调肝中药对VC不育大鼠性腺轴的调控，检测SCF/c-kit的表达，电镜观察睾丸组织各细胞超微结构的变化。在体外研究了调肝中药对支持细胞TM4的FSHR和SCF的影响；对生精细胞GC2的细胞周期及凋亡的影响。结果显示，VC模型组大鼠性腺轴各激素水平明显异常，其中睾丸组织中FSH、LH水平明显升高，FHSR水平明显降低，经调肝中药治疗后缓解了该异常变化。SCF/c-kit系统也出现异常，其中VC大鼠睾丸组织的SCF水平明显升高，c-kit水平明显降低，经调肝中药治疗后表达趋于正常。电镜观察 VC导致支持细胞和生精细胞超微结构损伤。体外实验表明调肝中药可以下调支持细胞SCF的表达，上调FSHR的表达，但对生精细胞的细胞周期及凋亡没有影响。本研究阐明了VC导致不育是通过性腺轴-SCF/c-kit通路实现的，调肝中药可以缓解该通路各分子的异常变化，是治疗VC的有效疗法。