鸡MyD88基因对巨噬细胞增殖和凋亡的影响

Effect of chicken MyD88 gene on macrophage proliferation and apoptosis

为探究鸡MyD88基因对巨噬细胞增殖和凋亡的影响,通过构建MyD88基因过表达质粒,以及合成MyD88基因干扰片段,并分别转染至鸡巨噬细胞(HD11)细胞中,采用CCK-8、流式细胞术及qRT-PCR方法检测过表达和干扰MyD88对HD11细胞增殖和凋亡的影响。结果显示,MyD88过表达可显著降低HD11细胞活力(P<0.05),下调促增殖标志基因CCND1和PCNA的表达水平(P<0.05),上调抑增殖标志基因P21的表达水平(P<0.05),减少S期细胞数,阻滞细胞周期进程,增加HD11细胞凋亡率,上调促凋亡基因Caspase 3、Caspase 9和Bax的表达水平(P<0.05)。干扰MyD88可显著提高HD11细胞活力(P<0.05),上调促增殖标志基因的表达水平,降低G1期细胞数,增加S期细胞数,加快细胞周期进程,降低HD11细胞凋亡率。结果表明,MyD88基因能抑制HD11细胞增殖,促进HD11细胞凋亡。

To investigate the effects of chicken MyD88 gene on macrophage proliferation and apoptosis,MyD88 gene overexpression plasmid was constructed,MyD88 gene interference fragments were synthesized and transfected into chicken macrophage(HD11)cells,respectively,and the effects of overexpression and interference of MyD88 on proliferation and apoptosis of HD11 cells were examined by CCK-8,flow cytometry and qRT-PCR.The results showed that MyD88 overexpression significantly decreased HD11 cell viability(P<0.05),down-regulated the expression levels of pro-proliferation marker genes CCND1 and PCNA(P<0.05),up-regulated the expression level of proliferation inhibitor marker gene P21(P<0.05),decreased the number of S phase cells,blocked cell cycle progression,increased the apoptosis rate of HD11 cells,and up-regulated the expression levels of pro-apoptotic genes Caspase 3,Caspase 9 and Bax expression levels(P<0.05).Interfering with MyD88 significantly increased HD11 cell viability(P<0.05),upregulated the expression level of pro-proliferation marker genes,decreased the number of G1 phase cells,increased the number of S phase cells,accelerated cell cycle progression and decreased the rate of HD11 cell apoptosis.The above results indicate that MyD88 gene inhibits HD11 cell proliferation and promotes HD11 cell apoptosis.