In recent years, resveratrol has been shown to protect against metabolic damage, including obesity-associated subfertility/infertility. In the present study, proteomic alterations in testicular tissues were investigated by tandem mass tag (TMT) in mice fed with a high-fat diet (HFD) without or with resveratrol supplementation (HFD+RSV). Serum testosterone levels, spermatozoa parameters and testicular histological morphology were assessed. Resveratrol treatment was shown to significantly reduce serum cholesterol, prevent the HFD-induced reductions in serum testosterone and spermatozoa parameters, and decrease the ultrastructural degeneration of testicular tissues. The comparative proteomics analysis revealed 58 differentially expressed proteins between the HFD and control groups and 38 differentially expressed proteins between the HFD and HFD+RSV groups. Gene ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the most highly enriched differential proteins were correlated to spermatozoa function and cholesterol metabolism. The real-time RT-PCR and western blotting results confirmed the differential expression of the corresponding proteins related to spermatozoa function that were identified by proteomics. The present study provides new insight into the mechanisms of the beneficial effects of resveratrol, and may present it as a potential therapeutic strategy for obesity-associated male subfertility/infertility.Abbreviations:TMT: Tandem mass tag; HFD: High-fat diet; RSV: Resveratrol; GO: Gene ontology; Protein-proteinKEGG: Kyoto Encyclopedia of Genes and Genomes; RT-PCR: Reverse transcription-polymerase chain reaction; SDS-PAGE: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis; PVDF: Polyvinylidene fluoride; ECL: Enhanced chemiluminescence; RIPA: Radio-immunoprecipitation assay; CTRL: Control; PPI: interaction; RIA: Radioimmunoassay; T: Testosterone; TG: Triglycerides; TC: Total cholesterol; LDL-c: Low-density lipoprotein cholesterol; HDL-c: High-density lipoprotein cholesterol; Crisp1: Cysteine-rich secretory protein 1; SIRT1: Sirtuin 1; GPx5: Glutathione peroxidase 5; Svs4: Seminal vesicle secretory protein 4; Tssk3: Testis-specific serine kinase 3; Pate4: Prostate and testis expressed 4; Sva: Seminal vesicle antigen; Lcn5: Lipocalin 5; Spinkl: Serine protease inhibitor, Kazal type-like.

中文翻译：

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比较蛋白质组学揭示了白藜芦醇对高脂饮食诱导的小鼠睾丸损伤的保护作用。

近年来，白藜芦醇已被证明可以抵抗代谢损害，包括与肥胖相关的不育/不育。在本研究中，通过串联质量标签（TMT）研究了饲喂高脂饮食（HFD）或未添加白藜芦醇（HFD + RSV）的小鼠睾丸组织中的蛋白质组学变化。评估血清睾丸激素水平，精子参数和睾丸组织学形态。白藜芦醇治疗可显着降低血清胆固醇，防止HFD诱导的血清睾丸激素和精子参数降低，并减少睾丸组织的超微结构变性。比较蛋白质组学分析显示，HFD和对照组之间有58个差异表达的蛋白质，HFD和HFD + RSV组之间有38个差异表达的蛋白质。基因本体论（GO）术语富集和京都基因与基因组百科全书（KEGG）途径分析显示，富集程度最高的差异蛋白与精子功能和胆固醇代谢相关。实时RT-PCR和蛋白质印迹结果证实了由蛋白质组学鉴定的与精子功能相关的相应蛋白质的差异表达。本研究为白藜芦醇的有益作用机理提供了新的见解，并可能将其作为肥胖相关的男性不育/不育症的潜在治疗策略。HFD：高脂饮食；RSV：白藜芦醇；GO：基因本体论；Protein-proteinKEGG：《京都基因与基因组百科全书》；RT-PCR：逆转录聚合酶链反应；SDS-PAGE：十二烷基硫酸钠-聚丙烯酰胺凝胶电泳；PVDF：聚偏二氟乙烯；ECL：增强化学发光；RIPA：放射免疫沉淀试验；CTRL：控制；PPI：互动；RIA：放射免疫分析；T：睾丸激素；TG：甘油三酸酯；TC：总胆固醇；LDL-c：低密度脂蛋白胆固醇；HDL-c：高密度脂蛋白胆固醇；Crisp1：富含半胱氨酸的分泌蛋白1；SIRT1：Sirtuin 1；GPx5：谷胱甘肽过氧化物酶5；Svs4：精囊分泌蛋白4；Tssk3：睾丸特异性丝氨酸激酶3；Pate4：前列腺和睾丸表示4；Sva：精囊泡抗原；Lcn5：Lipocalin 5；Spinkl：丝氨酸蛋白酶抑制剂，