中医学报2025，Vol.40Issue(5)：911-918,8.DOI:10.16368/j.issn.1674-8999.2025.05.146
五味扶正益精汤治疗男性不育症精子DNA损伤患者的蛋白质组学分析及初步实验验证
Proteomic Analysis and Preliminary Experimental Verification of Wuwei Fuzheng Yijing Decoction in Treatment of Male Infertility Patients with Sperm DNA Damage
摘要
Abstract
Objective:To analyze the proteomic characteristics of Wuwei Fuzheng Yijing Decoction in patients with sperm DNA damage in male infertility,and to carry out experimental verification based on sphingolipid metabolism pathway.Methods:A total of 20 patients with sperm DNA damage in male infertility were randomly divided into the experimental group and the verification group,with 10 cases in each group,and all of them were treated with Wuwei Fuzheng Yijing Decoction for 12 weeks.In addition,10 healthy subjects were used as the control group.Flow cytometry was used to detect the DNA fragmentation index(DFI)of the patient′s sperm.DIA pro-teomics technology was used to detect seminal plasma protein profile,and differentially expressed proteins were screened.The Kyoto En-cyclopedia of Genes and Genomes(KEGG)enrichment analysis of differentially expressed proteins was performed using the DAVID da-tabase.The contents of sphingomyelin phosphodiesterase 1(SMPD1),sphingosine-1-phosphate(S1P)and ceramide(Cer)were detected by ELISA.RT-qPCR was used to detect the expression levels of SMPD1,S1P and Cer genes in seminal plasma.Results:Com-pared with before treatment,the DFI level of sperm in the experimental group and the validation group decreased after treatment(P<0.001).DIA proteomics identified 2 753 quantitative proteins.Compared with the control group,there were 542 differentially expressed proteins in the seminal plasma in the experimental group before treatment(308 proteins were up-regulated and 234 proteins were down-regulated).Compared with before treatment,there were 505 differentially expressed proteins in the seminal plasma in the experi-mental group after treatment(267 proteins were up-regulated and 238 proteins were down-regulated).There were 235 intersecting proteins between the two.KEGG analysis showed that the differentially expressed proteins and intersecting proteins were concentrated and enriched in the sphingolipid metabolism pathway.ELISA showed that compared with the control group,the contents of SMPD1 and Cer in seminal plasma increased and the content of S1P decreased in the validation group before treatment(P<0.01),and compared with the content before treatment,the contents of SMPD1 and Cer in seminal plasma decreased and the content of S1P increased after treatment in the validation group(P<0.01).RT-qPCR showed that compared with the control group,the levels of SMPD1 mRNA and Cer mRNA in seminal plasma increased and the level of S1P mRNA decreased before treatment in the validation group(P<0.01),and compared with the pre-treatment group,the levels of SMPD1 mRNA and Cer mRNA in seminal plasma decreased and the level of S1P mRNA increased after treatment in the validation group(P<0.01).Conclusion:Wuwei Fuzheng Yijing Decoction can im-prove sperm DNA damage in male infertility patients,and its mechanism may be related to the regulation of sphingolipid metabolism pathway.关键词
五味扶正益精汤/男性不育症/精子DNA损伤/DNA碎片指数(DFI)/蛋白质组学/鞘脂代谢Key words
Wuwei Fuzheng Yijing Decoction/male infertility/sperm DNA damage/DNA fragmentation index(DFI)/proteomics/sphingolipid metabolism分类
中医学引用本文复制引用
马鹏,张琦,王祖龙..五味扶正益精汤治疗男性不育症精子DNA损伤患者的蛋白质组学分析及初步实验验证[J].中医学报,2025,40(5):911-918,8.基金项目
国家自然科学基金面上项目(82174377) （82174377）
河南省自然科学基金项目(242300420431) （242300420431）
