Assessment of Electroacupuncture Therapy with Distant-Approximal Acupoints Based on the HPT Axis in Rats with Oligoasthenospermia Through Transcriptomic Analysis.

Abstract

The transcriptomic analysis was used to explore the effect of electroacupuncture therapy with distant-approximal acupoints based on the hypothalamic-pituitary-testicular (HPT) on gene expression patterns and pathways in oligoasthenospermia (OAT) rats. In this study, the rat model of OAT after intragastric administration of adenine was selected as the research object, and randomly divided into a blank group (C), a model group (M), and a electroacupuncture therapy with distant-approximal acupoints group (D). After electroacupuncture intervention, the epididymal sperm quality and serum sex hormone levels of rats was detected and three tissue samples of HPT axis were taken, and differentially expressed genes (DEGs) were screened by transcriptome sequencing technology. GO functional annotation and KEGG pathway enrichment analysis were performed on the DEGs. The oxidative stress related indicators in serum and HPT axis were also detected to verify the transcriptomic analysis results. Compared with group C, group M rats showed a decrease in sperm count (p < 0.001), sperm survival rate (p < 0.001), and sperm motility rate (p < 0.001); the serum levels of GnRH in the group M rats decreased (p < 0.001), FSH increased (p < 0.001), LH increased (p < 0.001), and T decreased (p < 0.001). Compared with group M, group D rats showed an increase in sperm count (p < 0.01), sperm survival rate (p < 0.001), sperm motility rate (p < 0.001), an increase in GnRH levels (p < 0.001), a decrease in FSH levels (p < 0.01), a decrease in LH levels (p < 0.001), and an increase in T levels (p < 0.001). In bioinformatics analysis, compared with group M, we identified 1656, 518, 530 DEGs in the hypothalamus, pituitary, and testis in group D, respectively. Combining the go and KEGG analysis results, three oxidative stress signaling pathways that may be related to electroacupuncture intervention in OAT rats were screened. It mainly involves the glutamatergic synaptic pathway, the MAPK signaling pathway and the glutathione metabolism pathway. Six key genes (Gng12、Grin1、Gng7、Jun、Nf1 and Gstp1) were identified as key candidate genes regulating epididymal sperm quality on the HPT axis, which may affect the reproductive function of rats by affecting the process of oxidative stress in vivo. No matter in serum or in three tissues of HPT axis, GPX4 level in group M was decreased compared with Group K (p < 0.0001), while GPX4 level in group D was increased compared with group M (p < 0.0001). This study found that the effect of electroacupuncture therapy with distant-approximal acupoints based on the HPT axis in rats with OAT is related to the process of oxidative stress. And the main genes involved in the oxidative stress pathway were identified, which provided directions and ideas for subsequent research. But these results are only the preliminary results of transcriptomics, and relevant experiments need to be designed to further verify the mechanism of electroacupuncture therapy in rats with OAT.