【摘要】 目的:检测牙周炎患者自身组织核酸刺激小鼠巨噬细胞后炎性相关因子NOD样受体蛋白3（NLRP3）、半胱氨酸天冬氨酸特异蛋白酶1 （caspase-1）、白细胞介素1β（IL-1β）和白细胞介素18 （IL-18）mRNA表达水平,探讨牙周炎患者自身组织核酸对小鼠巨噬细胞中炎性相关因子的作用。方法:于慢性牙周炎患者翻瓣术中采集炎性牙周组织、于牙周组织健康患者冠延长术中采集健康牙周组织,分别提取组织总RNA逆转录成cDNA (cDNA质量浓度为1mg·L^-1)。培养小鼠巨噬细胞RAW264.7,分为对照组和实验组,分别加入上述提取的健康牙周组织cDNA （健康牙周组织）和炎症牙周组织cDNA （炎症牙周组织）共孵育。共孵育4、6和8h后,实时定量聚合酶链反应（RT-PCR）法检测各组RAW264.7细胞中NLRP3、caspase-1、IL-1β和IL-18mRNA表达水平。结果:显微镜下观察,小鼠巨噬细胞RAW264.7生长状态良好,细胞形态似圆形和多边形,胞浆透明,胞体丰满。与对照组比较,实验组在4、6和8h时RAW264.7细胞中NLRP3、caspase-1、IL-1β和IL-18mRNA表达水平明显升高（P<0.05或P<0.01）,6h时实验组RAW264.7细胞中NLRP3、caspase-1、IL-1β和IL-18mRNA表达水平最高。结论:牙周炎患者自身组织核酸可促进小鼠巨噬细胞中炎性相关因子mRNA表达,牙周炎患者自身组织核酸对小鼠巨噬细胞的活化具有免疫调节作用。更多还原

【Abstract】 Objective:To detect the mRNA expression levels of inflammatory-related factors NLRP3,caspase-1,IL-1β,and IL-18 in the murine macrophages infected by periodontitis patient’s own tissue nucleic acid,and to discuss the effects of periodontitis patient’s own tissue nucleic acid on the inflammation-related factors in the macrophages.Methods:The inflammatory periodontal tissue samples were collected during periodontal flap surgery of the chronic periodontitis patients,and the healthy periodontal tissue samples were collected from the patients without any periodontal diseases undergoing crown lengthening surgery.Then the total RNA from gingival tissue was extracted and reversely transcribed into cDNA.The cultured mouse macrophages RAW264.7 were divided into control group and experiment group,then the healthy periodontal tissue cDNA and inflammatory periodontal tissue cDNA(the cDNA at a concentration of 1 mg·L^-1)were added into the RAW264.7 cells,respectively.Real-time PCR was used to detect the mRNA expression levels of NLRP3,Caspase-1,IL-1β,and IL-18 in the macrophages in various groups at 4,6 and 8 hafter incubation.Results:The microscope observation showed that the mouse macrophages RAW264.7 grew well with round and polygon shapes,clear cytoplasm,and full cell body.Compared with control group,the expression levels of NLRP3,Caspase-1,IL-1β,and IL-18 mRNA in the RAW264.7 cells in experiment group at 4,6,and 8 hwere increased significantly（P<0.05 or P<0.01）,and the expression levels of NLRP3,Caspase-1,IL-1β,and IL-18 mRNA in RAW264.7 cells at 6 hin experiment group were the highest.Conclusion:The periodontitis patient’s own tissue nucleic acids can promote the mRNA expressions of inflammation-related factors in the RAW264.7 cells,suggesting that the periodontitis patient’s own tissue nucleic acid has an immunomodulatory effect on the activation of RAW264.7 cells.更多还原