肺炎支原体脂质相关膜蛋白诱发THP-1细胞炎症反应研究

A study on inflammation of THP-1 cells induced by lipid-associated membrane protein of Mycoplasma pneumoniae

探讨肺炎支原体脂质相关膜蛋白(LAMPs)诱发THP-1细胞炎症反应机制。提取LAMPs并刺激THP-1细胞,ELISA法检测细胞培养液上清中IL-1β,IL-18和TNF-α浓度,qRT-PCR法检测NLRP3和Caspase-1基因表达量,免疫印迹技术检测NF-κB信号通路活化,流式细胞术检测活性氧(ROS)生成量。结果显示:LAMPs显著提高IL-1β,IL-18和TNF-α表达(P<0.05);NLRP3和Caspase-1基因表达量均显著升高(P<0.05);LAMPs刺激后胞核NF-κB p65蛋白表达显著升高(P<0.05);细胞ROS生成量显著升高(P<0.05)。LAMPs可以通过ROS/NF-κB/NLRP3途径诱发宿主巨噬细胞炎症反应,且随浓度升高诱导炎症反应能力增强。

The purpose of the study is to explore the mechanism of Mycoplasma pneumoniae lipid-related membrane proteins(LAMPs)inducing the inflammatory response of THP-1 cells.LAMPs are extracted to stimulate THP-1 cells,detecting the concentration of IL-1β,IL-18 and TNF-αin the cell culture supernatant by ELISA and the expression of NLRP3 and Caspase-1 genes by qRT-PCR.The activation degree of NF-κB signaling pathway was determined by Western blotting,and the amount of reactive oxygen species(ROS)produced by flow cytometry is measured.The results showed that LAMPs significantly increased the expression of IL-1β,IL-18 and TNF-α(P<0.05).Both NLRP3 and caspase-1 gene expression increased significantly(P<0.05).The expression of nuclear NF-κB p65 protein was significantly increased after LAMPs stimulation(P<0.05).The amount of cellular ROS generation increased significantly(P<0.05).LAMPs can induce the inflammatory response of host macrophages through the ROS/NF-κB/NLRP3 pathway,and the ability to induce inflammatory response increases as the concentration increases.