■支持细胞对于维持精子发生和衰老过程中的功能下降至关重要。淫羊藿有补肾壮阳的功效,加强骨骼和肌肉,驱风驱湿,常用于治疗肾阳虚,阳痿和遗精。淫羊藿苷是淫羊藿的主要活性成分,具有延缓衰老和改善男性生殖功能障碍的作用。然而,目前尚不清楚淫羊藿苷如何通过保护睾丸支持细胞的连接功能免受损害来缓解年龄相关的睾丸功能下降.
目的:本研究旨在评价淫羊藿苷对支持细胞连接功能损伤的改善作用,并探讨其作用机制。
方法:采用雄性C57BL/6小鼠和小鼠睾丸支持细胞系TM4细胞评价淫羊藿苷对衰老相关的睾丸支持细胞连接功能损伤的改善作用。H&E染色,透射电子显微镜,qPCR,蛋白质印迹,分子对接,siRNA转染,在这项研究中进行了免疫荧光。
结果:淫羊藿苷的饮食给药显著减轻了年龄相关的生精功能恶化,睾丸重量和指数升高证明了这一点,精子浓度和精子活力。此外,淫羊藿苷保护睾丸支持细胞连接功能免受与年龄相关的损害,如睾丸支持细胞数量增加所证明的,改进的紧密连接超微结构,和上调连接相关蛋白(ZO-1,Occludin和β-Catenin)。此外,淫羊藿苷显著上调睾丸支持细胞ERα/c-fos信号和PKR通路。同样,体外研究表明,ERα的缺失,c-fos或PKR消除淫羊藿苷对支持细胞连接功能损伤的改善作用。
结论:淫羊藿苷通过ERα/c-fos信号上调PKR通路,减少支持细胞连接功能损伤,从而有效缓解年龄相关的睾丸功能下降。因此,通过ERα/c-fos信号上调PKR途径减轻睾丸支持细胞连接功能损伤可能是预防和治疗睾丸生精功能衰老的有效靶点。
UNASSIGNED: Sertoli cells are vital to maintain spermatogenesis and their function decline during aging. Epimedium has the effects of tonifying kidney-yang, strengthening bones and muscles, and expelling wind and dampness, and is commonly used in the treatment of kidney-yang deficiency, impotence and spermatorrhea.
Icariin is the main active ingredients from Epimedium exhibiting delaying aging effects and improving male reproductive dysfunction. Whereas, it remains poorly understood how icariin alleviates age-associated decline in testicular function by protecting against the damage of junction function of Sertoli cells.
OBJECTIVE: This study aimed to evaluate the improvement effect of
icariin on Sertoli cell junction function damage and explore the underlying mechanisms.
METHODS: Male C57BL/6 mice and mouse Sertoli cell line TM4 cells were utilized to assess the improvement effect of icariin on aging-associated Sertoli cell junction function injury. H&E staining, transmission electron microscopy, qPCR, Western blot, molecular docking, siRNA transfection, and immunofluorescence were performed in this study.
RESULTS: Dietary administration of
icariin remarkly attenuated age-associated deterioration in spermatogenic function as evidenced by elevated testicular weight and index, sperm concentration and sperm viability. In addition,
icariin protected Sertoli cell junction function from age-associated damage as proven by increased Sertoli cell numbers, improved tight junction ultrastructure, and upregulated junction-related proteins (ZO-1, Occludin and β-Catenin). Moreover,
icariin significantly upregulated ERα/c-fos signaling and PKR pathway in testicular Sertoli cells. Similarly, in vitro studies revealed that deletion of ERα, c-fos or PKR abolished the improvement effects of icariin on Sertoli cell junction function damage.
CONCLUSIONS: Icariin effectively mitigates age-associated decline in testicular function by diminished Sertoli cell junction function damage through upregulating PKR pathway via ERα/c-fos signaling. Therefore, attenuating Sertoli cell junction function injury by the upregulation of PKR pathway via ERα/c-fos signaling probably indicates an effective target for the prevention and treatment of testicular spermatogenic function with aging.