目的:研究人牙髓成纤维细胞(human dental pulp fibroblasts,HDPF)中炎症体NLRP3、Caspase-1基因和蛋白的表达和相关调控因素。方法:分别利用免疫细胞化学染色和RT-PCR检测NLRP3、Caspase-1、P2X7、IL-β基因和NLRP3、Caspase-1蛋白在正常HDPF中的表达情况;利用荧光实时定量PCR检测变异链球菌和ATP刺激后,HDPF中NLRP3、Caspase-1、P2X7、IL-β基因表达的变化。结果:NLRP3、Caspase-蛋白和NLRP3、Caspase-1、P2X7、IL-β基因在正常HDPF中均有表达;HDPF经ATP和变异链球菌共同刺激后,NLRP3、Caspase-1、P2X7、IL-β mRNA表达水平均明显升高,与对照组相比,差异均有统计学意义(P〈0.05);而单独ATP或变异链球菌刺激后,各基因的表达水平变化不明显(P〉0.05)。结论:在正常HDPF中存在着NLRP3、Caspase-1炎症体通路;ATP和变异链球菌共同刺激能调节NLRP3、Caspase-1炎症体通路。
AIM: To investigate the mRNA and protein expression of inflammasome NLRP3 and Caspase- 1, as well as their regulatory factors in human dental pulp fibroblasts (HDPFs). METHODS: The protein expression of NLRP3 and Caspase-1 in HDPF was detected by immunocytochemistry. The mRNA expression of NLRP3, Caspase -1, P2X7 and IL-1β was detected by semi-quantitative RT-PCR and quantitative Real-time RT-PCR. RESULTS: NLRP3 and Caspase-1 proteins were observed in HDPFs. The expressions of NLRP3, Caspase-1 , P2X7 and IL-1β mRNA were significanty up-regulated by ATP and Streptococcus mutans co-stimulation(P 〈 0.05 ). However , single stimulation with ATP or Streptococcus mutans could not produce a significant effect on the relevant gene expression (P 〉 0.05). CONCLUSION: The inflammasome pathway of NLRP3 and Caspase-1 is present in HDPFs and it can be activated by co-stimulation of ATP and Streptococcus mutans.