重度冷应激对三河牛血液生化指标及相关基因表达的影响

Effects of Severe Cold Stress on Blood Biochemical Parameters and Related Gene Expression in Sanhe Cattle

本研究旨在检测与牛冷应激反应相关的生化指标,筛选参与牛冷应激反应的相关基因。以内蒙古三河牛为研究对象,对30头青年牛进行室外-32℃冷应激处理3h,采集冷应激前后试验个体血样,检测血液生化指标及基因表达量的变化。冷应激3h后,试验个体T3、T4及ACTH3个血液生化指标极显著升高(P<0.01)。mRNA差异显示方法(DDRT)检测到基因LIN54和KLF12在冷应激后表达量降低,荧光定量RT-PCR验证了冷应激后2个基因表达下调(LIN54下调显著(P<0.05),KLF12下调极显著(P<0.01))的结果。筛选到的2个基因分别属于LINC和KLF家族,LINC参与有丝分裂基因的活化,KLF参与细胞增殖和凋亡。T3、T4及ACTH 3项生化指标可以初步作为衡量三河牛冷应激反应强弱的侯选指标在大群体中进行验证,筛选到的2个基因,可作为参与三河牛冷应激反应的重要侯选基因,进一步研究2个基因在牛冷应激反应中的作用机制,综合这些生化指标及侯选基因表达的变化,用于评价个体冷应激反应的强度,在三河牛抗冷应激育种中将具有更广泛的应用价值。

The aim of this study was to detect the biochemical indexes and relevant genes related to cold stress in Sanhe cattle.Thirty heifer Sanhe cattle in Inner Mongolia were kept at-32 ℃for 3h,and then blood samples were collected before and after cold stress separately to detect the biochemical indexes and gene expression changes.After 3hcold stress,T3,T4 and ACTH inblood significantly increased(P<0.01).The expressions of LIN54 and KLF12decreased in silver staining map produced by mRNA differential display method(DDRT),decrease of the 2genes expression were further verified at the level of P<0.05 for LIN54and P<0.01 for KLF12by fluorescence quantitative RT-PCR.The 2genes belonged to LINC and KLF families,respectively,LINC involved in mitotic gene activation,KLF involved in cell proliferation and apoptosis.T3,T4 and ACTH may be used as the candidate indexes to measure the strength of cold stress in large population,LIN54 and KLF12may be the important candidate genes for further study on genetic mechanism of cold stress in cattle.The combination of these biochemical indexes and expression change of candidate genes would be used to assess the change of individual under cold stress,which will be more valuable in breeding for cold resistance ability in Sanhe cattle.