Zhu Congxu, Li Yingqiu, Liu Lumei, Xie Jing, Hu Zongren, He Qinghu, Sheng Wen
Andrology Laboratory, Hunan University of Chinese Medicine, Changsha, 410208, China.
School of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha, 410208, China.
Sci Rep. 2025 Jul 28;15(1):27498. doi: 10.1038/s41598-025-09667-8.
Oligoasthenozoospermia (OAS) is a common cause of male infertility, and Guilu Erxian Glue (GLEXG) is a traditional Chinese medicine known to improve male reproductive function. The role of GLEXG in OAS and its mechanism of action are not well understood. After the OAS model was established with Tripterygium wilfordii polyglycoside (GTW)-induced mice, the mice were orally administered GLEXG. The effects of GLEXG on testis and epididymis of GTW-induced mice were evaluated by HE, TEM, and Johnson scores. Meanwhile, the effects of GLEXG on semen quality of GTW-induced mice were explored by CTC staining and sperm viability assay, etc. Additionally, an H2O2-treated mouse spermatogonial (GC-2) cell model was established and incubated with GLEXG-containing serum. The mechanism of action of GLEXG was investigated by immunofluorescence, methylation-specific PCR (MSP), and western blot. The OAS mouse model was successfully constructed, and the testes, epididymis, and semen quality of mice were improved after GLEXG treatment. In vivo, GLEXG could promote the expression of TDRP, cAMP, and PKA, and the methylation levels of TDRP genes. Mechanistically, GLEXG could improve sperm viability in HO-treated GC-2 cells by promoting the methylation level of TDRP and TDRP genes, and also improve the rate of competent spermatozoa in HO-treated GC-2 cells by promoting cAMP/PKA. Our study demonstrates that GLEXG can improve sperm quality in GTW-induced mice and H2O2-treated GC-2 cells by promoting TDRP methylation and modulating the cAMP/PKA pathway. We elucidated the regulatory mechanisms of GLEXG in OAS, providing new potential targets for the treatment of OAS.
少弱精子症（OAS）是男性不育的常见原因，而龟鹿二仙胶（GLEXG）是一种已知可改善男性生殖功能的传统中药。GLEXG在OAS中的作用及其作用机制尚不清楚。在用雷公藤多苷（GTW）诱导的小鼠建立OAS模型后，给小鼠口服GLEXG。通过苏木精-伊红（HE）染色、透射电子显微镜（TEM）和约翰逊评分评估GLEXG对GTW诱导小鼠睾丸和附睾的影响。同时，通过计算机辅助精液分析（CTC）染色和精子活力测定等方法探索GLEXG对GTW诱导小鼠精液质量的影响。此外，建立了过氧化氢（H2O2）处理的小鼠精原细胞（GC-2）细胞模型，并用含GLEXG的血清进行孵育。通过免疫荧光、甲基化特异性聚合酶链反应（MSP）和蛋白质免疫印迹法研究GLEXG的作用机制。成功构建了OAS小鼠模型，GLEXG治疗后小鼠的睾丸、附睾和精液质量得到改善。在体内，GLEXG可促进睾丸决定相关蛋白（TDRP）、环磷酸腺苷（cAMP）和蛋白激酶A（PKA）的表达以及TDRP基因的甲基化水平。机制上，GLEXG可通过促进TDRP和TDRP基因的甲基化水平提高H2O2处理的GC-2细胞中的精子活力，还可通过促进cAMP/PKA提高H2O2处理的GC-2细胞中的合格精子率。我们的研究表明，GLEXG可通过促进TDRP甲基化和调节cAMP/PKA途径改善GTW诱导小鼠和H2O2处理的GC-2细胞中的精子质量。我们阐明了GLEXG在OAS中的调控机制，为OAS的治疗提供了新的潜在靶点。
