肾上腺素对猪季节性基因表达的影响

选用部分具有季节性表达的基因,探讨肾上腺素对这些基因mRNA表达量的影响。(1)将猪肾细胞(PK15)分别置于含有100,250,500,750,1 000 nmol/L肾上腺素的培养基中培养6 h,实时荧光定量PCR(qRT-PCR)检测ADRβ2、ARNTL、CLOCK、IL-6R、NR3C1、VDR的mRNA表达量的变化。结果表明:1 000 nmol/L肾上腺素处理PK15细胞时,与对照组相比ADRβ2、IL-6R的mRNA表达量明显升高,差异极显著(P0.01);VDR的mRNA表达量降低,差异极显著(P0.01);CLOCK、NR3C1、ARNTL的mRNA表达量无明显变化。(2)选择1 000 nmol/L的肾上腺素分别处理PK15细胞6,12,24,36,48 h。qRT-PCR检测ADRβ2的mRNA表达量变化。结果表明:与对照组相比,培养12 h时ADRβ2的mRNA表达量达到1.6倍,差异极显著(P0.01),随着培养时间延长,ADRβ2的mRNA表达量逐渐降低,36 h时降到最低,为对照组的0.4倍,差异显著(P0.05)。这说明肾上腺素对部分季节周期性变化的基因具有调节作用,季节变化产生应激时可以通过肾上腺素影响ADRβ2、IL-6R、VDR的表达,从而影响机体功能。

Effects of Epinephrine on Expression Level of Porcine Seasonal Gene

The aim of this study is to investigate the effects of epinephrine on the expression levels of partial seasonal gene mRNA.Porcine kidney cells were maintained in DMEM supplemented with different concentrations of epinephrine for 6 h,with the concentration gradient of 100,250,500,750,1 000 nmol/L,respectively.qRT-PCR assay was proceeded to detect the expression level of seasonal gene mRNA,including ADRβ2,ARNTL,CLOCK,IL-6R,NR3C1 and VDR.The results of qRT -PCR showed that when 1 000 nmol/L epinephrine was sued to treat PK15 cells,the expression levels of ADRβ2 and IL-6R mRNA were significantly up-regulated compared with the control group.But the expression level of VDR mRNA was significantly down-regulated and the expression levels of CLOCK,NR3C1 and ARNTL mRNA had no significant difference.1 000 nmol/L epinephrine was chosen to treat PK15 cells for 6 h,12 h,24 h,36 h and 48 h,respectively,and then the expression level of ADRβ2 mRNA was analyzed by qRT-PCR.The qRT-PCR results showed that the expression level of ADRβ2 mRNA after being cultured for 12 h was 1.6 times as much as that of the control group.With the increase of incubation time,the expression level of ADRβ2 mRNA was gradually attenuated,reaching the minimum at 36 h,which is 0.4 times as much as that of the control group.The study demonstrates that epinephrine plays an important role in the partial gene characterized by seasonal changes.The seasonal changes produced by stress influence the expression levels of ADRβ2,IL-6R and VDR,and affect the function of organism by regulating the level of epinephrine.