Zhao Han, Mou Qiao, Wang Fang, Du Zhi-Qiang, Yang Cai-Xia
College of Animal Science and Technology, Yangtze University, Jingzhou, Hubei 434025, China.
College of Animal Science and Technology, Yangtze University, Jingzhou, Hubei 434025, China.
J Steroid Biochem Mol Biol. 2024 Oct;243:106580. doi: 10.1016/j.jsbmb.2024.106580. Epub 2024 Jul 10.
Vitamin C (Ascorbic acid, AA), as vital micro-nutrient, plays an essential role for male animal reproduction. Previously, we showed that vitamin C reprogrammed the transcriptome and proteome to change phenotypes of porcine immature Sertoli cells (iSCs). Here, we used LC-MS-based non-targeted metabolomics to further investigate the metabolic effects of vitamin C on porcine iSCs. The results identified 43 significantly differential metabolites (DMs) (16 up and 27 down) as induced by vitamin C (L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate, AA2P) treatment of porcine iSCs, which were mainly enriched in steroid related and protein related metabolic pathways. ELISA (Enzyme-Linked ImmunoSorbent Assay) showed that significantly differential metabolites of Dehydroepiandrosterone (DHEA) (involved in steroid hormone biosynthesis) and Desmosterol (involved in steroid degradation) were significantly increased, which were partially consistent with metabolomic results. Further integrative analysis of metabolomics, transcriptomics and proteomics data identified the strong correlation between the key differential metabolite of Dehydroepiandrosterone and 6 differentially expressed genes (DEGs)/proteins (DEPs) (HMGCS1, P4HA1, STON2, LOXL2, EMILIN2 and CCN3). Further experiments validated that HMGCS1 could positively regulate Dehydroepiandrosterone level. These data indicate that vitamin C could modulate the metabolism profile, and HMGCS1-DHEA could be the pathway to mediate effects exerted by vitamin C on porcine iSCs.
维生素C（抗坏血酸，AA）作为一种重要的微量营养素，对雄性动物繁殖起着至关重要的作用。此前，我们发现维生素C可重编程转录组和蛋白质组，从而改变猪未成熟支持细胞（iSCs）的表型。在此，我们使用基于液相色谱-质谱联用的非靶向代谢组学方法，进一步研究维生素C对猪iSCs的代谢影响。结果鉴定出43种由维生素C（L-抗坏血酸2-磷酸倍半镁盐水合物，AA2P）处理猪iSCs诱导产生的显著差异代谢物（DMs）（16种上调和27种下调），这些代谢物主要富集在类固醇相关和蛋白质相关的代谢途径中。酶联免疫吸附测定（ELISA）表明，参与类固醇激素生物合成的脱氢表雄酮（DHEA）和参与类固醇降解的羊毛甾醇的显著差异代谢物显著增加，这与代谢组学结果部分一致。对代谢组学、转录组学和蛋白质组学数据的进一步综合分析确定了脱氢表雄酮的关键差异代谢物与6个差异表达基因（DEGs）/蛋白质（DEPs）（HMGCS1、P4HA1、STON2、LOXL2、EMILIN2和CCN3）之间的强相关性。进一步的实验验证了HMGCS1可以正向调节脱氢表雄酮水平。这些数据表明维生素C可以调节代谢谱，并且HMGCS1-DHEA可能是介导维生素C对猪iSCs作用的途径。
