College of Animal Science, Yangtze University, Jingzhou, 434025, Hubei, China; College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, Heilongjiang, China.
College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, Heilongjiang, China.
Theriogenology. 2022 Aug;188:13-21. doi: 10.1016/j.theriogenology.2022.05.011. Epub 2022 May 17.
Somatic Sertoli cells constitute the microenvironment and produce essential substances, to support male germ cell development and maturation in testis. We previously found that ascorbic acid treatment of porcine immature Sertoli cells enhances its proliferation and secretion of reproductive hormones and metabolites, and reprograms the global transcriptome. Proteomics is a powerful tool to systematically profile the underlying protein changes. Here, by employing the TMT-based quantitative proteomics method, we identified 96 and 64 significantly up- and down-regulated proteins in porcine immature Sertoli cells treated by ascorbic acid, respectively. Gene enrichment (GO and KEGG) and protein-protein interaction analyses revealed important molecular pathways (dioxygenase activity, sterol biosynthetic process, PI3K-Akt, negative regulation of peptide hormone secretion, extracellular matrix etc.). Further validation of three proteins, HMGCS1 (cholesterol synthesis), P4HA1 (glycolysis) and KDM5A (demethylation of histone 3 at lysine 4), confirmed their significant differential abundance, respectively. Taken together, our findings show that ascorbic acid can alter multiple important protein molecules and related signaling pathways, which could explain partially phenotypic changes (proliferation, apoptosis, nucleic acid methylation, lactate and reproductive hormone secretion) of porcine immature Sertoli cells as induced by ascorbic acid.
体 Sertoli 细胞构成了微环境，并产生必需的物质，以支持睾丸中男性生殖细胞的发育和成熟。我们之前发现，抗坏血酸处理猪未成熟 Sertoli 细胞可增强其增殖和生殖激素及代谢物的分泌，并对其进行了全局转录组重编程。蛋白质组学是一种系统分析潜在蛋白变化的有力工具。在这里，我们采用基于 TMT 的定量蛋白质组学方法，分别鉴定出 96 种和 64 种在抗坏血酸处理的猪未成熟 Sertoli 细胞中显著上调和下调的蛋白质。基因富集（GO 和 KEGG）和蛋白质-蛋白质相互作用分析揭示了重要的分子途径（双加氧酶活性、固醇生物合成过程、PI3K-Akt、肽激素分泌的负调控、细胞外基质等）。进一步验证了三种蛋白质，即 HMGCS1（胆固醇合成）、P4HA1（糖酵解）和 KDM5A（组蛋白 3 赖氨酸 4 的去甲基化），它们的丰度差异具有统计学意义。综上所述，我们的研究结果表明，抗坏血酸可以改变多个重要的蛋白质分子和相关信号通路，这可以部分解释抗坏血酸诱导的猪未成熟 Sertoli 细胞的表型变化（增殖、凋亡、核酸甲基化、乳酸和生殖激素分泌）。
