Vitamin E is generally believed to promote the production of ovine sperm mainly through its antioxidant effect. Our previous studies have shown that some non-antioxidant genes may also be key in mediating this process. The objective of this study was to identify key candidate proteins that were differentially expressed in response to a treatment with Vitamin E. Prepubertal ovine testicular cells were isolated and divided into two groups. They were either treated with 800 μM Vitamin E (based on our previous results) or used as a non-treated control. After 24 h, all the cells were harvested for proteomic analysis. We found 115 differentially expressed proteins, 4 of which were up-regulated and 111 were down-regulated. A GO term enrichment analysis identified 127 Biological Process, 63 Cell Component and 26 Molecular Function terms that were enriched. Within those terms, 13, 11 and 26 terms were significantly enriched, respectively. Terms related to membrane and enzyme activity including the inner acrosomal membrane, signal peptidase complex, cysteine-type endopeptidase activity, etc., were also markedly enriched, while none of the KEGG pathways were enriched. We found that many of the differentially expressed proteins, such as CD46 (membrane cofactor protein), FLNA (Filamin A), DYSF (Dysferlin), IFT20 (Intraflagellar transport 20), SPCS1 (Signal peptidase complex subunit 1) and SPCS3 (Signal peptidase complex subunit 3) were related to the acrosomal and plasma membranes. A parallel reaction monitoring (PRM) analysis verified that Vitamin E improved spermatogenesis by regulating the expression of FLNA, SPCS3, YBX3 and RARS, proteins that are associated with the plasma membranes and protamine biosynthesis of the spermatozoa.

一般认为维生素E主要通过其抗氧化作用来促进绵羊精子的产生。我们以前的研究表明，某些非抗氧化剂基因也可能是介导这一过程的关键。这项研究的目的是鉴定在应答维生素E后差异表达的关键候选蛋白。将青春期前的睾丸细胞分离并分为两组。他们要么用800μM维生素E治疗（根据我们之前的结果），要么用作未治疗的对照。24小时后，收获所有细胞用于蛋白质组学分析。我们发现了115个差异表达的蛋白质，其中4个被上调，而111个被下调。GO术语富集分析确定了127个生物过程，63个细胞成分和26个分子功能术语被富集。在这些术语中，分别显着丰富了13、11和26个术语。与膜和酶活性有关的术语，包括内顶体膜，信号肽酶复合物，半胱氨酸型内肽酶活性等，也显着丰富，而KEGG途径均未丰富。我们发现许多差异表达的蛋白质，例如CD46（膜辅因子蛋白），FLNA（纤维蛋白A），DYSF（Dysferlin），IFT20（鞭毛内转运20），SPCS1（信号肽酶复合物亚基1）和SPCS3（信号肽酶）复杂的亚基3）与顶体和质膜有关。平行反应监测（PRM）分析证明，维生素E通过调节FLNA，SPCS3，YBX3和RARS的表达来改善精子发生，