脂联素对酮病奶牛中性粒细胞自噬相关基因mRNA表达的影响

Effect of adiponectin on mRNA expression levels of autophagy related gene in neutrophil of ketosis cows

通过模拟向高BHBA的中性粒细胞培养液中添加不同质量浓度的脂联素(ADPN),运用荧光定量PCR技术,分别检测中性粒细胞自噬相关基因mRNA的表达水平。结果显示:添加高BHBA(4.8mol/L)的中性粒细胞培养6h后,与对照组相比,自噬相关基因LC3B、Beclin、ULK1的mRNA表达水平显著升高(P<0.01),SQSTM1(p62)、mTOR的mRNA表达水平显著下降(P<0.01);添加不同质量浓度的ADPN(25,50,100μg/L)后,相对于高BHBA组,中性粒细胞自噬相关基因LC3B、Beclin、ULK1的mRNA表达水平显著下调,且在ADPN 50μg/L时,具有极显著差异性(P<0.01);此外SQSTM1、mTOR的mRNA表达水平显著升高。结果表明:BHBA可增加中性粒细胞自噬相关基因的表达,ADPN可抑制高BHBA引起的中性粒细胞细胞程序性死亡,进一步说明ADPN对调控酮病奶牛中性粒细胞免疫抑制具有重要的价值意义。

The aim of this study was to evaluate the effect of adiponctin on the neutrophil pro- grammed cell death（autophagy） of cows. By adding different concentrations of ADPN to the cul- ture medium with a high concentration of BHBA, detect the mRNA expression levels of autoph- agy related genes using the technology of quanlititative PCR. The results showed that treatments with high levels of BHBA （4.8 mol/L） cultured for 6 h, significantly elevated the mRNA expres- sion levels of LC3B,Beclinl ,ULK1 in neutrophil,compared with the control group（P〈0. 01） ;the mRNA expression levels of mTOR and SQSTM1 were significantly decreased（P〈0.05）. After adding diferent concentrations of ADPN（25,50 and 100 μg/L）, compared with the high BHBA, the mRNA expression levels of LC3B,Beclin and ULK1 in neutrophil were significantly lowered, especilly at ADPN 50 μg/L （P〈0.01） ;in additon, the mRNA expression levels of SQSTM1 and roTOR were significantly increased（P（0.05）. In summary,ADPN has an improvement effects on the neutro- phils programmed cell death induced by high levels of BHBA in cows, which further illustrates the impor- tant valuation of ADPN on regulating immunosuppression of neutroohils in ketosis cows.