Rig-I对LPS诱导的巨噬细胞增殖、凋亡及功能的作用研究

Effect of Rig-I on Proliferation, Apoptosis and Function of LPS-induced Macrophages

该研究旨在探讨维甲酸诱导基因I(retinoic acid-inducible gene I,Rig-I)对脂多糖(lipopolysaccharide,LPS)诱导的巨噬细胞增殖、凋亡和细胞因子分泌等生物学功能的影响及其作用机制。以不同剂量的LPS刺激Rig-I基因沉默和过表达的小鼠巨噬细胞系Raw264.7细胞,采用CCK-8和流式细胞术检测细胞的生长状态,采用q PCR方法检测细胞因子TNF-α、IL-10、IL-1和IL-6相关基因的表达水平,并采用Western blot方法检测LPS诱导的TLR4信号通路的相关蛋白表达水平。CCK-8和流式细胞术结果显示,Rig-I促进巨噬细胞的增殖并抑制LPS诱导的细胞凋亡;q PCR结果表明,Rig-I促进巨噬细胞中TNF-α、IL-10、IL-1和IL-6基因的表达。进一步的实验证实,Rig-I通过激活AKT及其下游蛋白p-38、NF-κB和Bcl-x L等抑制细胞凋亡并促进细胞因子的表达。该研究首次证实了Rig-I可通过AKT信号通路对LPS诱导的巨噬细胞增殖、凋亡及功能进行调节。

To investigate the biology function and mechanism of Rig-I on lipopolysaccharide（LPS）-induced macrophages such as proliferation, apoptosis and cytokines secretion, macrophages（Raw264.7） with Rig-I gene knock-down or overexpression were treated with different doses of LPS for indicated times. The viability was analyzed by CCK-8 and apoptosis was measured by flow cytometry. The expression levels of related cytokines were examined by q PCR. The Toll like receptor-4（TLR4） signaling induced by LPS was detected by Western blot. CCK-8 and flow cytometry analyses showed that Rig-I promoted the proliferation of macrophages and inhibited LPSinduced apoptosis. q PCR results indicated that Rig-I upregulated the expression of TNF-α, IL-10, IL-1 and IL-6 in macrophage cells. These effects caused by Rig-I were further demonstrated by activation of AKT and its downstream p-38, NF-κB and Bcl-x L. These data firstly suggested that Rig-I could regulate proliferation, apoptosis and function of LPS-induced macrophages through AKT signaling pathway.