奶牛白细胞氧代谢VEGF通路和新基因散热调节

VEGF pathway of oxygen metabolism and regulation of heat dissipation of new gene in white blood cells of dairy cows

旨在讨论奶牛适应高温散热生理基因调控和信号通路。采集热环境(25,30,35℃)中娟珊牛血液样品,分离白细胞提取总RNA,采用Smart-Seq2扩增、构建序列文库,RNA-seq高通量测序筛选基因和q-PCR荧光定量进行验证。结果显示热差异基因功能主要集中在免疫抑制、氧化应激和内分泌紊乱;qRT-PCR验证PRDM1、SAMSN1、BHLHE40、NR4A2、DUSP6、EDN1、MYL9、PTGS2和IL-8基因表达上调,而IL1A和TGFB2基因下调;VEGF信号通路调节氧代谢,发现新基因ENSBTAG00000005339与PRDM1、DUSP67、SAMSN1、END1、ENSBTAG00000010303、PTGS2和IL-1参与VEGF信号通路。结果显示,奶牛白细胞中VEGF信号通路及其基因(ENSBTAG00000005339和DUSP6、DUSP67、SAMSN1、END1、ENSBTAG00000010303、PTGS2和IL-1A)调控氧代谢实现高温散热。

Aimed to explore cow adaptation to hot environment by physiological gene regulation and the signal pathway.Jersey cows were natural exposed to different temperature of 25,30 and 35℃.Leukocyte(5×10~6)was isolated from whole blood sample(2 mL)for mRNA-seq analysis and then amplified via the Smart-Seq2 method to construct RNA libraries and sequenced using RNA-seq,further verified using q-PCR.Transcriptome analysis results showed that VEGF pathway manipulated oxidative stress of leukocyte of cow which would enhance heat loss at high temperature.And a novel gene ENSBTAG00000005339 at VEGF pathway was relative with PRDM1,DUSP67,SAMSN1,END1,ENSBTAG00000010303,PTGS2 and IL-1 A;some relative genes were validated using qRT-PCR and gene expression of PRDM1,SAMSN1,BHLHE40,NR4 A2,DUSP6,EDN1、MYL9、PTGS2 and IL-8 increased mRNA expression,but TGFB2 and IL1 A decreased in the experiment group.Thus,transcriptome analysis revealed that VEGF pathway and genes(ENSBTAG00000005339,DUSP6,DUSP67,SAMSN1,END1,ENSBTAG00000010303,PTGS2 and IL-1 A)in leukocyte were as potential markers for oxidative stress assessment at dairy cattle.