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目的观察肺炎支原体(Mycoplasma pneunoniae,Mp)感染RAW264.7细胞早期NLRP3炎性体及前炎性细胞因子的表达。方法将RAW264.7细胞随机分为5组,正常组用常规方法培养,不感染Mp;4个实验组RAW264.7细胞均用Mp感染4h,感染复数(细胞︰Mp)分别为1︰20、1︰40、1︰80、1︰100,采用FQ-PCR法检测细胞NLRP3、ASC、caspase-1mRNA表达和IL-1β、IL-18水平。结果 Mp感染复数1︰20、1︰40、1︰80、1︰100组NLRP3、ASC、caspase-1mRNA表达量分别为2.10±0.62、2.14±0.66、2.66±0.69、3.29±0.64和3.91±0.83、4.21±0.95、4.25±0.86、4.30±0.99和1.65±0.48、1.65±0.36、1.94±0.51、2.00±0.57,与正常对照组0.98±0.08、1.00±0.08、0.99±0.09比较,差异均有统计学意义(P<0.01);感染复数1︰100组IL-1β为200.00±9.25pg/ml,与对照组159.92±5.89pg/ml比较差异有统计学意义(P<0.01)。结论 Mp感染可诱导RAW264.7细胞NLRP3炎性体活化。NLRP3炎性体可能参与了Mp的早期感染。
Objective To investigate the expression of NLRP3 inflammasome and proinflammatory cytokines in RAW264.7 cells infected with Mycoplasma pneunoniae (Mp). Methods RAW264.7 cells were randomly divided into 5 groups. The normal group was cultured by conventional methods without infection of Mp. The RAW264.7 cells in 4 experimental groups were all infected with Mp for 4 hours, the multiplicity of infection (cells: Mp) were 1: 20, 1: 40,1: 80,1: 100. The expression of NLRP3, ASC, caspase-1 mRNA and IL-1β and IL-18 were detected by FQ-PCR. Results The mRNA expression levels of NLRP3, ASC and caspase-1 in the 1: 20,1: 40,1: 80,1: 100 group infected with Mp were 2.10 ± 0.62, 2.14 ± 0.66, 2.66 ± 0.69, 3.29 ± 0.64 and 3.91 ± 0.83 , 4.21 ± 0.95, 4.25 ± 0.86, 4.30 ± 0.99 and 1.65 ± 0.48, 1.65 ± 0.36, 1.94 ± 0.51 and 2.00 ± 0.57, respectively, which were statistically different from those in normal control group (0.98 ± 0.08,1.00 ± 0.08,0.99 ± 0.09) (P <0.01). The IL-1β in the 1: 100 infected group was 200.00 ± 9.25 pg / ml, which was significantly different from that in the control group (159.92 ± 5.89 pg / ml) (P <0.01). Conclusion Mp infection can induce the activation of NLRP3 inflammasome in RAW264.7 cells. The NLRP3 inflammasome may be involved in the early Mp infection.