五子衍宗丸对少精症小鼠睾丸基因表达谱的影响

Effects of Wuzi Yanzong pills on gene expression profile of testis in mice with oligospermia

目的观察五子衍宗丸对少精症小鼠睾丸基因表达谱的影响,从基因表达层面探讨五子衍宗丸对少精症的调控机制。方法采用白消安法建立少精症小鼠动物模型(n=24),少精症小鼠随机分成少精症组(生理盐水,14 mL/kg/d,灌胃2周,n=12),五子衍宗丸治疗组(五子衍宗丸,1.56 g/kg/d,灌胃2周,n=12),另设Balb/c小鼠为正常对照组(生理盐水,14 mL/kg/d,灌胃2周,n=12),给药结束后取各组睾丸组织,提取总RNA,采用基因芯片技术检测基因表达谱,筛选五子衍宗丸调控的差异基因,通过基因本体(GO)注释分析及京都基因和基因组百科全书(KEGG)信号通路分析,对五子衍宗丸调控的差异基因进行功能注释和信号通路分析。结果在睾丸组织基因表达谱中,与少精症组相比,五子衍宗丸治疗组和正常对照组共同调控的差异表达基因共有597个,其中五子衍宗丸治疗组和正常对照组同时上调的基因有55个,同时下调的基因542个。GO功能分析发现五子衍宗丸调控的睾丸差异基因(597)显著富集的生物学过程有1110条,涉及精子生成、生精细胞减数分裂的周期、精子细胞分化等生物过程;KEGG通路分析发现五子衍宗丸调控的睾丸差异基因(597)显著调节的信号通路共有109条,包括同源重组、肾素-血管紧张素系统、胆固醇代谢信号通路等。结论五子衍宗丸通过影响少精症小鼠睾丸组织的基因表达,调节生精细胞的减数分裂、睾丸同源重组、肾素-血管紧张素系统、胆固醇代谢等途径,修复睾丸生精阻滞。

Objective To observe the effect of Wuzi Yanzong pills on gene expression profile of tesis of mice with oligospermia,and to explore the regulation mechanism of W uzi Yanzong pills on oligospermia from the perspective of gene expression.Methods The mouse model with oligospermia was established by busulfan method(n=24),mice with oligospermia were randomly divided into oligospermia group(normal saline,14 mL/kg/d,gavage for 2 weeks,n=12),Wuzi Yanzong pills treatment group(Wuzi Yanzong pills,1.56g/kg/d,gavage for 2 weeks,n=12),and Balb/c mice as the normal control group(normal saline,14 mL/kg/d,gavage for 2 weeks,n=12).After drug administration,testis tissue was taken from each group and total RNA were extracted.Gene expression profile was detected by using gene chip technology,and the different genes regulated by Wuzi Yanzong pills were screened out.Through ontology(GO)annotation analysis and Kyoto Encyclopedia of Genes and Genomics(KEGG)signaling pathway analysis,functional annotation and signal pathway analysis were performed on the differential genes regulated by regulated by Wuzi Yanzong pills.Results In the gene expression profile of testis tissue,compared with the oligospermia group,there were 597 differentially expressed genes co-regulated by the Wuzi Yanzong pills treatment group and the normal control group,among which 55 genes were up-regulated and 542 genes were down-regulated in the Wuzi Yanzong pills treatment group and the normal control group.GO functional analysis revealed that there were 1110 biological processes,involving sperm generation,spermatogenic meiosis cycle,sperm cell differentiation and other biological processes regulated by Wuzi Yanzong pills.KEGG pathway analysis revealed that the differential testis genes(597)regulated by Wuzi Yanzong pills significantly regulated 109 signaling pathways,including homologous recombination,renin-angiotensin system,cholesterol metabolism signaling pathway and others.Conclusion Wuzi Yanzong pills can regulate the meiosis of spermatogenic cells,homologous recombination of testis,renin-angiotensin system,cholesterol metabolism pathway and others,so as to repair the blocking of testicular spermatogenesis,by affecting the gene expression of testis tissue in mice with oligospermia.