Abstract：
The present study was conducted to understand transcriptional response of skin fibroblast of yak (Bos grunniens) and cows of Bos indicus origin to hypoxia stress. Six primary fibroblast cell lines derived from three individuals each of Ladakhi yak (Bos grunniens) and Sahiwal cows (Bos indicus) were exposed to low oxygen concentration for a period of 24 h, 48 h and 72 h. The expression of 10 important genes known to regulate hypoxia response such as HIF1A, VEGFA, EPAS1, ATP1A1, GLUT1, HMOX1, ECE1, TNF-A, GPx and SOD were evaluated in fibroblast cells of Ladakhi yak (LAY-Fb) and Sahiwal cows (SAC-Fb) during pre- and post-hypoxia stress. A panel of 10 reference genes (GAPDH, RPL4, EEF1A1, RPS9, HPRT1, UXT, RPS23, B2M, RPS15, ACTB) were also evaluated for their expression stability to perform accurate normalization. The expression of HIF1A was significantly (p < 0.05) induced in both LAY-Fb (2.29-fold) and SAC-Fb (2.07-fold) after 24 h of hypoxia stress. The angiogenic (VEGFA), metabolic (GLUT1) and antioxidant genes (SOD and GPx) were also induced after 24 h of hypoxia stress. However, EPAS1 and ATP1A1 induced significantly (p < 0.05) after 48 h whereas, ECE1 expression induced significantly (p < 0.05) at 72 h after exposure to hypoxia. The TNF-alpha which is a pro-inflammatory gene induced significantly (p < 0.05) at 24 h in SAC-Fb and at 72 h in LAY-Fb. The induction of hypoxia associated genes indicated the utility of skin derived fibroblast as cellular model to evaluate transcriptome signatures post hypoxia stress in populations adapted to diverse altitudes.
摘要：
进行本研究是为了了解牦牛（Bosgrunniens）和Bosindicus牛的皮肤成纤维细胞对缺氧应激的转录反应。来自Ladakhi牦牛（Bosgrunniens）和Sahiwal牛（Bosindicus）的三个个体的六个原代成纤维细胞细胞系暴露于低氧浓度24小时，48h和72h。已知10个调节缺氧反应的重要基因如HIF1A、VEGFA,EPAS1,ATP1A1,GLUT1,HMOX1,ECE1,TNF-A,在缺氧前后应激期间，在拉达基牦牛（LAY-Fb）和Sahiwal奶牛（SAC-Fb）的成纤维细胞中评估了GPx和SOD。一组10个参考基因(GAPDH，RPL4,EEF1A1,RPS9,HPRT1,UXT,RPS23,B2M,还评估了RPS15，ACTB）的表达稳定性以进行准确的归一化。在低氧胁迫24小时后，在LAY-Fb（2.29倍）和SAC-Fb（2.07倍）中均显着（p<0.05）诱导了HIF1A的表达。血管生成（VEGFA），低氧胁迫24h后还诱导了代谢（GLUT1）和抗氧化基因（SOD和GPx）。然而,EPAS1和ATP1A1在48小时后显著诱导(p<0.05)，暴露于缺氧后72小时，ECE1表达显着诱导（p<0.05）。作为促炎基因的TNF-α在SAC-Fb中在24小时和在LAY-Fb中在72小时显著诱导(p<0.05)。缺氧相关基因的诱导表明皮肤来源的成纤维细胞作为细胞模型用于评估适应不同海拔高度的群体中缺氧应激后的转录组特征。
