摘要
为探究健康和酮病奶牛CD4+T细胞中丝氨酸代谢相关基因的表达及炎症相关基因的表达.首先使用磁珠分选的方法,分离奶牛外周血中CD4+T细胞.其次,使用荧光定量PCR方法检测健康、酮病奶牛CD4+T细胞中丝氨酸代谢相关基因 PHGDH、PAST1、PSPH、SHMT1、SHMT2、SDS、SFXN1 及炎症相关基因 IL-6、IFN-γ、IL-17A、FOXP3、IL-10、TGF-β的表达.与健康奶牛相比,酮病奶牛CD4+T细胞中PHGDH、PAST1和PSPH的转录水平均极显著升高(P＜0.01),SDS的转录水平与翻译水平均显著升高(P＜0.01).而SHMT2的转录水平显著降低(P＜0.05).SFXN1的转录水平与翻译水平均显著降低(P＜0.05).SHMT1的转录水平降低但无显著性.与健康奶牛相比,酮病奶牛CD4+T细胞中IL-6、IFN-γ、IL-17A的转录水平均极显著升高(P＜0.01),IL-10、TGF-β、FOXP3的转录水平均显著降低(P＜0.05).结果表明,酮病奶牛丝氨酸合成过程基因表达升高,丝氨酸分解过程相关基因表达降低,而促炎因子表达升高,抑炎因子表达降低,说明丝氨酸代谢在奶牛体内发生炎症过程中发挥重要作用.
Abstract
To investigate the expression of serine metabolism related genes and inflammatory relat-ed genes in CD4+T cells of healthy and cows with ketosis.Firstly,CD4+T cells in peripheral blood of dairy cows were isolated by magnetic bead sorting.Second,expression of serine metabolism re-lated genes PHGDH,PAST1,PSPH,SHMT1,SHMT2,SDS,SFXN1 and inflammation related genes IL-6,IFN-γ,IL-17A,FOXP3,IL-10,TGF-β in CD4+T cells in healthy and ketosis cows was detected by fluorescence quantitative PCR.The transcription levels of PHGDH,PAST1 and PSPH in CD4+T cells of ketosis cows were significantly increased compared with healthy cows(P＜0.01),the transcription level and translation level of SDS were significantly increased(P＜0.01).The transcription levels of SHMT2 were significantly decreased(P＜0.05).Transcription and translation levels of SFXN1 were significantly decreased(P＜0.05).The transcription level of SHMT1 was decreased but not significantly.Compared with healthy cows,the transcription levels of IL-6,IFN-γ and IL-17A in CD4+T cells of ketosis cows were significantly increased(P＜0.01),the transcription levels of IL-10,TGF-β and FOXP3 were significantly decreased(P＜0.05).The results showed that the expression of genes related to serine synthesis increased,the expression of genes related to serine decomposition decreased,the expression of pro-inflammatory factors increased,and the expression of anti-inflammatory factors decreased,suggesting that serine metab-olism plays an important role in the inflammatory process of dairy cows.
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