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基于转录组学分析评估远近配穴电针疗法对少弱精子症大鼠下丘脑-垂体-睾丸轴的影响

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

作者信息

Hongyu Li, Nan Yang, Kaiying Li, Zhenning Zhao, Lili Zhao, Jing Mu, Huisheng Ma

机构信息

Ningxia Medical University, Yinchuan, 750004, Ningxia Hui Autonomous Region, China.

General Hospital of Ningxia Medical University, Yinchuan, 750004, Ningxia Hui Autonomous Region, China.

出版信息

Reprod Sci. 2025 Apr;32(4):1228-1240. doi: 10.1007/s43032-025-01821-x. Epub 2025 Feb 27.

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.

摘要

采用转录组学分析方法,探讨基于下丘脑-垂体-睾丸(HPT)轴的远近配穴电针疗法对少弱精子症(OAT)大鼠基因表达模式和通路的影响。本研究选取灌胃腺嘌呤后建立的OAT大鼠模型为研究对象,随机分为空白组(C)、模型组(M)和远近配穴电针治疗组(D)。电针干预后,检测大鼠附睾精子质量和血清性激素水平,采集HPT轴的三个组织样本,采用转录组测序技术筛选差异表达基因(DEG)。对DEG进行GO功能注释和KEGG通路富集分析。同时检测血清和HPT轴中的氧化应激相关指标,以验证转录组学分析结果。与C组相比,M组大鼠精子数量(p<0.001)、精子存活率(p<0.001)和精子活动率(p<0.001)均降低;M组大鼠血清GnRH水平降低(p<0.001),FSH升高(p<0.001),LH升高(p<0.001),T降低(p<0.001)。与M组相比,D组大鼠精子数量(p<0.01)、精子存活率(p<0.001)、精子活动率(p<0.001)升高,GnRH水平升高(p<0.001),FSH水平降低(p<0.01),LH水平降低(p<0.001),T水平升高(p<0.001)。生物信息学分析显示,与M组相比,D组下丘脑、垂体和睾丸中分别鉴定出1656、518、530个DEG。结合GO和KEGG分析结果,筛选出三条可能与电针干预OAT大鼠氧化应激相关的信号通路。主要涉及谷氨酸能突触通路、MAPK信号通路和谷胱甘肽代谢通路。确定了6个关键基因(Gng12、Grin1、Gng7、Jun、Nf1和Gstp1)为HPT轴上调节附睾精子质量的关键候选基因,它们可能通过影响体内氧化应激过程来影响大鼠的生殖功能。无论在血清还是HPT轴的三个组织中,M组的GPX4水平均低于K组(p<0.0001),而D组的GPX4水平高于M组(p<0.0001)。本研究发现,基于HPT轴的远近配穴电针疗法对OAT大鼠的治疗作用与氧化应激过程有关。并鉴定出氧化应激通路中的主要相关基因,为后续研究提供了方向和思路。但这些结果仅是转录组学的初步结果,需要设计相关实验进一步验证电针治疗OAT大鼠的机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c75d/11978543/9d32704bc6ae/43032_2025_1821_Fig1_HTML.jpg

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