Guilu erxian glue improves sperm quality in oligasthenospermia mice by promoting TDRP methylation and regulating the cAMP/PKA pathway.

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 H₂O₂-treated GC-2 cells by promoting the methylation level of TDRP and TDRP genes, and also improve the rate of competent spermatozoa in H₂O₂-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.