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目的:观察芩百清肺浓缩丸含药血清对肺炎支原体(Mycoplasma pneumoniae,MP)感染RAW264.7细胞NLRP3炎性体及IL-1β的影响。方法:试验分为空白对照组、模型组和芩百清肺浓缩丸含药血清组。常规培养RAW264.7细胞、肺炎支原体菌株,制备芩百清肺浓缩丸含药血清,用10 MOI MP感染RAW264.7细胞,分别于8、16、24 h收集细胞。FQ-PCR检测NLRP3、ASC、Caspase-1 mRNA表达;Western-blot检测NLRP3、ASC、Caspase-1 p20蛋白表达;ELISA检测细胞上清液IL-1β含量。结果:与空白对照组比较,MP感染RAW264.7细胞后8、16、24 h NLRP3、ASC、Caspase-1 mRNA表达显著上调(P<0.05或P<0.01);感染后16、24 h NLRP3、ASC、Caspase-1 p20蛋白表达显著上调(P<0.05或P<0.01);感染后24 h细胞上清液IL-1β含量显著升高(P<0.01)。与模型组比较,芩百清肺浓缩丸含药血清可显著下调MP感染RAW264.7细胞16、24 h NLRP3、ASC、Caspase-1 mRNA的表达(P<0.05或P<0.01);显著降低感染后24 h NLRP3、ASC、Caspase-1p20蛋白表达和细胞上清液IL-1β的分泌(P<0.05或P<0.01)。结论:芩百清肺浓缩丸抗肺炎支原体作用的机制可能与下调NLRP3炎性体表达有关。
Objective: To observe the effect of Qinbaiqing Pneumoconiosis-containing serum on NLRP3 inflammasome and IL-1β in RAW264.7 cells infected with Mycoplasma pneumoniae (MP). Methods: The experiment was divided into blank control group, model group and Qin Bai Qingfei Pill containing drug serum group. RAW264.7 cells and Mycoplasma pneumoniae strains were routinely cultured, and serum containing Qinlian Qingfei Pill was prepared. RAW264.7 cells were infected with 10 MOI MP, and the cells were harvested at 8, 16 and 24 hours respectively. The expression of NLRP3, ASC and Caspase-1 mRNA was detected by FQ-PCR. The expressions of NLRP3, ASC and Caspase-1 p20 protein were detected by Western-blot. The content of IL-1β in supernatant was detected by ELISA. RESULTS: Compared with the blank control group, the mRNA expression of NLRP3, ASC and Caspase-1 in MP infected RAW264.7 cells was significantly up-regulated at 8, 16 and 24 h after MP infection (P <0.05 or P <0.01) (P <0.05 or P <0.01). The content of IL-1β in supernatant of cells was significantly increased at 24 h after infection (P <0.01). Compared with the model group, Qinbaiqing Pneumoconiosis-containing serum could significantly reduce the mRNA expression of NLRP3, ASC and Caspase-1 (P <0.05 or P <0.01) at 16 and 24 h in MP-infected RAW264.7 cells and significantly reduce the infection The expression of NLRP3, ASC, Caspase-1p20 protein and the secretion of IL-1β in cell supernatant (P <0.05 or P <0.01) after 24 h. Conclusion: The mechanism of Qinbaiqing Pneumoconiosis Pill against Mycoplasma pneumoniae may be related to the down-regulation of NLRP3 inflammasome expression.