Abstract
Objective:Exploring the mechanism of curcumin based on the RIPK1/RIPK3/MLKL signaling pathway in necroptosis of human gingival fibroblasts(HGFs)induced by Porphyromonas gingivalis lipopolysaccharide(LPS P.gingiva-lis)and its effects on a periodontitis rat model.Methods:HGF-1 cells were treated with different concentrations(0,5,10,15,20,40 μmol/L)of curcumin.Cell survival rate was detected by CCK-8 method,apoptosis rate was detected by flow cytometry,and necrotic apoptosis rate of HGFs stimulated by LPS P.gingivalis was detected by ELISA.The expression of interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α),and hypoxia inducible factor-1α(HIF-1α)in the RIPK1/RIPK3/MLKL pathway was also analyzed by immunofluorescence and Western blot.The phosphorylation levels of key proteins in the RIPK1/RIPK3/MLKL pathway were determined.Establish a rat periodontitis model induced by LPS P.gingivalis,and use Micro-CT to detect bone density(BMD),bone volume fraction(BV/TV),trabecular separation(Tb.Sp),and trabecular num-ber(Tb.N).RT-qPCR was used to detect the levels of IL-1β,IL-6,TNF-α and IL-10 mRNA in periodontal tissue,and West-ern blot was used to detect the phosphorylation levels of RIPK1/RIPK3/MLKL in rat periodontal tissue.Cell experiments were conducted with Control group,Model group,Curcumin group,RIPK activator TSZ group(TSZ group),and RIPK inhibi-tor Nec-1 group(Nec group).Animal experiments were conducted with Control group,Model group,Curcumin group,RIPK activator TSZ group(TSZ group),and positive drug Metronidazole(Met group).Results:After treatment with curcumin,the apoptosis rate,IL-1β,IL-6,TNF-α,HIF-1α levels,and RIPK1,RIPK3,and MLKL phosphorylation levels were all reduced(P<0.01).In animal models,after treatment with curcumin,BMD,BV/TV,and Tb.N increased,while Tb.Sp,IL-6,IL-1β,TNF-α,and IL-10 decreased(P<0.05).And the phosphorylation levels of RIPK1/RIPK3/MLKL in periodontal tissue de-creased(P<0.01).The effect was comparable to that of the Nec-1 inhibitor and the positive control drug Met,and it can an-tagonize the effects of TSZ activators.Conclusion:Curcumin alleviates periodontitis by targeting and inhibiting the RIPK1/RIPK3/MLKL signaling pathway,thereby blocking LPS P.gingivalis-induced necroptosis in HGFs and the inflammatory cas-cade reaction,effectively reducing periodontal tissue destruction and bone loss,providing a novel mechanistic basis for the prevention and treatment of periodontitis.关键词
姜黄素/RIPK1/RIPK3/MLKL/牙周炎Key words
Curcumin/RIPK1/RIPK3/MLKL/Periodontitis分类
医药卫生
