脓肿分枝杆菌经TLR2信号通路诱导人单核细胞株表达炎症细胞因子分析

Study on the inflammatory cytokines of human monocyte cell in Mycobacterium abscessus inducing through TLR2 pathway

目的了解脓肿分枝杆菌脓肿亚种和马赛亚种诱导人单核细胞株内TNF-α、IL-8和IL-10的表达变化及与TLR2信号途径的相关性。方法将脓肿分枝杆菌脓肿亚种和马赛亚种感染人单核细胞,感染复数(MOI)=3,采用荧光定量PCR检测人单核细胞被两细菌亚种感染6 h^18 h后胞内TNF-α、IL-8和IL-10的表达水平;采用AntiTLR2 Abs阻断TLR2受体,荧光定量PCR检测人单核细胞被感染6 h后胞内TNF-α、IL-8和IL-10的表达变化。结果脓肿亚种和马赛亚种作用人单核细胞6 h^18 h后,均可诱导人单核细胞内TNF-α、IL-8和IL-10的表达水平上调,以TNF-α表达水平上调明显,IL-10表达水平上调较低,差异有统计学意义(P<0.05);TLR2受体阻断后,脓肿亚种和马赛亚种感染人单核细胞上调胞内TNF-α、IL-8和IL-10表达均出现明显抑制,差异均有统计学意义(P<0.05)。结论脓肿分枝杆菌脓肿亚种和马赛亚种均可经TLR2信号途径诱导人单核细胞内TNF-α、IL-8和IL-10表达水平上调,以TNF-α表达水平上调明显。

Objective To determine the TNF-α,IL-8 and IL-10 expression levels induced by the M. abscessus subsp. abscessus（ Mabc） and Mycobacterium subsp. massiliense（ Mmass） and the correlation with the pathway of Toll-like receptor（ TLR）-2. Methods THP-1 cells were stimulated with Mabc and Mmass（ MOI = 3）. QRT-PCR was used to detect the TNF-α,IL-8 and IL-10 expression levels in 6-18 hours infected with Mabc and Mmass. TNF-α,IL-8 and IL-10 expression activity were measured by qRT-PCR with anti-TLR2 Abs in 6 hours infected with Mabc and Mmass. Results Mabc-induced TNF-α,IL-8 and IL-10 expression levels and Mmass-induced TNF-α,IL-8 and IL-10 expression levels were upregulated in the THP-1 cells within 6 h-18 h,the TNF-α expression level was significantly upregulated,the IL-10 expression level was slight upgegulated,with the differences statistically significant（ P 〈 0. 05）; after TLR2 receptor blockade,the expressions of TNF-α,IL-8 and IL-10 were significantly inhibited in the cells of the infected and infected human monocytes,with the differences statistically significant（ P 〈 0. 05）. Conclusion The TNF-α,IL-8 and IL-10 expression levels were upregulated through TLR2 pathway in the THP-1 cells infected with Mabc and Mmass,with the TNF-α expression level upregulation significant.