摘要
目的:研究敲低耐酸因子F-ATPase促进变形链球菌感染人牙髓成纤维细胞(human dental pulp fibroblasts,HDPFs)成骨/成牙本质分化的作用。方法:培养HDPFs并采用免疫荧光染色、流式细胞术进行鉴定,而后分为对照组、感染组、感染+si-NC组、感染+si-F-ATPase组、感染+si-F-ATPase+NLR家族PYRIN域蛋白3(NLRP3)激动剂(NIG)组。比较各组间白介素(IL)-1β、IL-18、肿瘤坏死因子-α(TNF-α)、NLRP3、ASC、Cleaved caspase-1、骨钙素(OCN)、Runt相关转录因子2(Runx2)以及成骨诱导分化后茜素红染色OD540nm值、碱性磷酸酶(ALP)活力的差异。结果:感染组HDPFs中IL-1β、IL-18、TNF-α的含量,NLRP3、ASC、Cleaved caspase-1的表达量均高于对照组,OCN、Runx2表达量及成骨诱导后的OD540nm水平、ALP活力均低于对照组;感染+si-F-ATPase组HDPFs中IL-1β、IL-18、TNF-α的含量,NLRP3、ASC、Cleaved caspase-1的表达量均低于感染组,OCN、Runx2表达量及成骨诱导后的OD540nm水平、ALP活力均高于感染组;感染+si-F-ATPase+NIG组HDPFs中IL-1β、IL-18、TNF-α的含量,NLRP3、ASC、Cleaved caspase-1的表达量均高于感染+si-F-ATPase组,OCN、Runx2表达量及成骨诱导后的OD540nm水平、ALP活力均低于感染+si-F-ATPase组。结论:敲低F-ATPase能够促进变形链球菌感染HDPFs的成骨/成牙本质分化,抑制NLRP3炎症小体介导的炎症反应是可能的分子机制。
Objective:To investigate the effect of knockdown of F-ATPase on promoting osteogenic/odontogenic differentiation of human dental pulp fibroblasts(HDPFs)infected by Streptococcus mutans.Methods:HDPFs were cultured and identified by immunofluorescence staining and flow cytometry.Then they were divided into control group,infection group,infection+si-NC group,infection+si-F-ATPase group,infection+si-F-ATPase+NLRP3 agonist(NIG)group.Then IL-1β,IL-18,TNF-α,NLRP3,ASC,Cleaved caspase-1,osteocalcin(OCN),Runt related transcription factor 2(Runx2)and alizarin red staining OD540 nm,ALP activity after osteogenic differentiation were compared.Results:The content of IL-1β,IL-18 and TNF-α,the expression of NLRP3,ASC and cleaved caspase-1 in HDPFs of infection group were higher than control group,the expression of OCN,Runx2,the level of OD540 nm and the activity of ALP after osteogenesis induction were lower than control group.The content of IL-1β,IL-18 and TNF-α,the expression of NLRP3,ASC and cleaved caspase-1 in HDPFs of infection+si-F-ATPase group were lower than infection group,the expression of OCN,Runx2,the level of OD540 nm and the activity of ALP after osteogenesis induction were higher than infection group;the content of IL-1β,IL-18 and TNF-α.The expression of NLRP3,ASC and cleaved caspase-1 in HDPFs of infection+si-F-ATPase+NIG group were higher than infection+si-F-ATPase group,the expression of OCN,Runx2,the level of OD540 nm and the activity of ALP after osteogenesis induction were lower than infection+si-F-ATPase group.Conclusion:Knockdown of F-ATPase can improve the osteogenic/odontogenic differentiation of Streptococcus mutans infected HDPFs,the inhibion of inflammation mediated by NLRP3 inflammasomes is the possible molecular mechanism.
作者
李亚
熊洁
LI Ya;XIONG Jie(Dongfeng Stomatological Hospital,Hubei University of Medicine,Hubei Shiyan 442000,China)
出处
《临床口腔医学杂志》
2021年第3期136-141,共6页
Journal of Clinical Stomatology
