Pax-8基因抑制后心肌细胞中UCP2的表达

目的 建立Pax-8基因敲除模型及Pax -8基因的体外干扰实验,探索心肌细胞中Pax-8基因抑制后解偶联蛋白2(uncouplingprotein 2,UCP2)基因表达的变化.方法 Pax-8基因敲除纯合子小鼠(Pax-8 KO-/-)为实验组,Pax-8基因敲除小鼠杂合子(Pax -8 KO+/-)和野生型(Pax-8KO+/+)为对照组;同时在体外实验的大鼠心肌细胞株中,加入Pax -8基因的小干扰RNA(Pax-8siRNA)为实验组,非特异性siRNA(NC siRNA)组和空白组为对照组.采用RT-PCR和Westernblotting技术测定UCP2的mRNA和蛋白质表达.结果 在Pax-8基因敲除动物模型中,与Pax-8基因敲除小鼠杂合子组和野生型组相比,实验组Pax-8基因敲除小鼠纯合子心脏组织UCP2的mRNA和蛋白表达均上调( P<0.05),而对照组之间差异无统计学意义(P>0.05);在体外干扰实验中的结果显示,与NC siRNA组和空白对照组比较,Pax-8siRNA组UCP2的mRNA表达和蛋白表达均上调(P<0.05),而NC siRNA组与空白对照组间差异无统计学意义(P>0.05).结论 Pax-8基因被抑制后,心肌细胞凋亡增加,UCP2基因表达上调,提示UCP2参与了心肌细胞凋亡的调控.

The expression of UCP2 gene in the myocardium after the inhibition of Pax-8 gene

Objective:To establish Pax-8 gene knockout model and H9C2(2-1) interference in vitro to inhibit Pax-8 gene in the myocardium,and explore the expression of uncoupling protein 2(UCP2) gene.Methods: Pax-8 knockout mouse homozygous(Pax-8 KO-/-) was built as the experimental group,Pax-8 knockout mouse heterozygous(Pax-8 KO+/-) and wild-type(Pax-8 KO+/+) group as control.In vitro experiments at the same time,short interference RNA targeting Pax-8(Pax-8 siRNA) was added into the H9C2(2-1) as experimental group,and non specific siRNA group(NC siRNA)and the blank group as control.RT-PCR and Western blotting were used to analyze the expressions of UCP2 mRNA and protein.Results: Compared to the Pax-8 knockout mice heterozy-gous and wild-type group,UCP2 mRNA and protein expression in Pax-8 knockout mice homozygous group were apparently increased(both P <0.05),while the difference in control group was not statistically significant(P >0.05).In comparison with NC siRNA group and blank control group,the expression of UCP2 mRNA and protein in Pax-8 siRNA group were both upregulated(both P <0.05),while difference in statistically no significant was found between NC siRNA group and blank control group(P >0.05).Conclusion: The apoptosis of myocar-dium is in-creased and the UCP2 expression upregulated after Pax-8 gene is inhibited,All the result indicate that UCP2 gene may participate in the regulation of the myocardial apoptosis.