紫檀芪调节AMPK/NF-κB/NLRP3信号通路对人牙周膜成纤维细胞炎症及凋亡的影响OA

Objective:To investigate the impacts of pterostilbene on inflammation and apoptosis of human periodontal ligament fibroblasts(PDLFs)by regulating the AMP-activated protein kinase(AMPK)/nuclear factor-kappa B(NF-κB)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)signaling pathway.Methods:Human PDLFs cells were divided into six group:control group,lipopolysaccharide(LPS)group,different-dose(5,10,and 20 μmol/L)pterostilbene groups,and pterostilbene(20 μmol/L)+AMPK inhibitor(1 μmol/L Compound C)group.After 2 h of pretreatment with pterostilbene and Compound C,the cells were cultured in medium containing 1 mg/L LPS for 24 h.CCK-8 assay was used to detect the viability of PDLFs cells.ELISA was applied to determine interleukin(IL)-6,tumor necrosis factor α(TNF-α),and IL-1β levels in PDLFs.Flow cytometry was adopted to detect apoptosis in PDLFs.RT-qPCR was utilized to measure the mRNA expression of AMPK,NF-κB,and NLRP3 in PDLFs.Western blotting was performed to observe the expression of AMPK,p-AMPK,NF-κB,p-NF-κB,and NLRP3 proteins in PDLFs.Results:Compared with the control group,the OD450 value,AMPK mRNA,and p-AMPK/AMPK of PDLFs in LPS group reduced,while the IL-6,TNF-α,IL-1β,apoptosis rate,NF-κB and NLRP3 mRNA,p-NF-κB/NF-κB and NLRP3 proteins increased.Compared with the LPS group,the OD450 value,AMPK mRNA,and p-AMPK/AMPK of PDLFs in Pterostilbene groups(5,10,and 20 μmol/L)increased,while the IL-6,TNF-α,IL-1β,apoptosis rate,NF-κB and NLRP3 mRNA,p-NF-κB/NF-κB and NLRP3 proteins reduced.Compared with the 20 μmol/L Pterostilbene group,the OD450 value,AMPK mRNA,and p-AMPK/AMPK of PDLFs in pterostilbene+Compound C group reduced,while the IL-6,TNF-α,IL-1β,apoptosis rate,NF-κB and NLRP3 mRNA,p-NF-κB/NF-κB and NLRP3 proteins increased.Conclusion:Pterostilbene activates AMPK to downregulate NF-κB and NLRP3,thereby inhibiting inflammation and apoptosis of PDLFs cells.