炎症环境下牙髓成纤维细胞中NOD样受体蛋白3炎症体相关分子表达的调节  

Regulating the expression of nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome-related molecules in dental pulp fibroblasts under inflammation

作　　者：张安生 张海欧 倪龙兴 Zhang Ansheng;Zhang Haiou;Ni Longxing(Department of Stomatology,Xi’an International Medical Center Hospital,Xi’an 710018,Shaanxi Province,China;Department of Ophthalmology and Otorhinolaryngology,the 928th Hospital of Chinese PLA,Haikou 570100,Hainan Province,China)

机构地区：[1]西安国际医学中心医院口腔科,陕西省西安市710018 [2]解放军联勤保障部队第928医院五官科,海南省海口市570100

出　　处：《中国组织工程研究》2022年第26期4107-4112,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:NOD样受体蛋白3炎症体是细胞抵御外来病原入侵的重要炎性因子,前期研究已证实其在牙髓成纤维细胞中有表达,然而炎症环境下牙髓成纤维细胞NOD样受体蛋白3炎症体相关分子的转录调节机制尚不清楚。目的:阐明炎症环境下牙髓成纤维细胞中NOD样受体蛋白3炎症体相关分子的转录调节机制。方法:选择第4代牙髓成纤维细胞,分6组:A组正常培养,不进行任何处理;B组加入脂多糖刺激6 h;C组加入Toll样受体4特异性抑制剂处理1 h,再加入Toll样受体4特异性抑制剂与脂多糖混合溶液处理6 h;D组加入髓样分化因子88特异性抑制剂处理1 h,再加入髓样分化因子88特异性抑制剂与脂多糖混合溶液处理6 h;E组加入核因子κB特异性抑制剂处理1 h,再加入核因子κB特异性抑制剂与脂多糖混合溶液处理6 h;F组加入髓样分化因子88特异性抑制剂阴性对照处理1 h,再加入髓样分化因子88特异性抑制剂阴性对照与脂多糖混合溶液处理6 h。采用RT-PCR检测NOD样受体蛋白3、Caspase-1与白细胞介素1βmRNA的表达,Western blot检测NOD样受体蛋白3与Caspase-1蛋白表达,ELISA法检测白细胞介素1β释放水平。结果与结论:①与A组比较,B组NOD样受体蛋白3、Caspase-1及白细胞介素1βmRNA表达水平升高(P<0.05);与B组比较,C-E组NOD样受体蛋白3、白细胞介素1βmRNA表达水平降低(P<0.05),C组Caspase-1 mRNA表达水平降低(P<0.05);②与A组比较,B组NOD样受体蛋白3、Caspase-1蛋白表达升高(P<0.05);与B组比较,C-E组NOD样受体蛋白3蛋白表达降低(P<0.05),C组Caspase-1蛋白表达降低(P<0.05);③与A组比较,B组白细胞介素1β水平升高(P<0.05);与B组比较,C-E组白细胞介素1β水平降低(P<0.05);④结果表明,脂多糖通过TLR4/MyD88/NF-κB信号通路上调牙髓成纤维细胞内NOD样受体蛋白3和白细胞介素1β的表达。BACKGROUND:Nucleotide-binding oligomerization domain(NOD)-like receptor protein 3 inflammasome is an important inflammatory factor for cellular defense against various pathogens,which has been confirmed to be expressed in dental pulp fibroblasts.However,the mechanisms underlying the transcription regulation of NOD-like receptor protein 3 inflammasome in dental pulp fibroblasts under inflammation remains unclear.OBJECTIVE:To elucidate the mechanisms underlying the transcription regulation of NOD-like receptor protein 3 inflammasome in dental pulp fibroblasts under inflammation.METHODS:The 4^(th) generation dental pulp fibroblasts were divided into six groups.Group A was cultured normally without any treatment.Group B was simulated with lipopolysaccharide for 6 hours.Group C was treated with a specific inhibitor of Toll-like receptor 4 for 1 hour,and then it was also treated with a mixed solution of Toll-like receptor 4 specific inhibitor and lipopolysaccharide for 6 hours.Group D was treated with a specific inhibitor of myeloid differentiation factor 88 for 1 hour,and then treated with a mixed solution of specific inhibitor of myeloid differentiation factor 88 and lipopolysaccharide for 6 hours.Group E was treated with a specific inhibitor of nuclear factor κB for 1 hour,and then treated with a mixture solution of specific inhibitor of nuclear factor κB and lipopolysaccharide for 6 hours.Group F was treated with a negative control of myeloid differentiation factor 88 specific inhibitor for 1 hour,and then treated with a mixed solution of myeloid differentiation factor 88 specific inhibitor negative control and lipopolysaecharide for 6 hours.Real-time polymerase chain reaction was used to detect the mRNA expression of NOD-like receptor protein 3,Caspase-1 and interleukin 1β.Western bolt was used to detect the expression of NOD-like receptor protein 3 and Caspase-1.And enzyme linked immunosorbent assay was used to detect the release level of interleukin 1β.RESULTS AND CONCLUSION:Compared with group A,the mRNA ex

关 键 词：牙髓成纤维细胞 固有免疫 NOD样受体4 NOD样受体蛋白3炎症体 信号通路 白细胞介素1β 核因子κB 

分 类 号：R459.9[医药卫生—治疗学] R781.3[医药卫生—临床医学]