Effect of acacetin on lipopolysaccharide induced apoptosis of dental pulp cells by regulating the HMGB1/TLR4 signaling pathway
- VernacularTitle:金合欢素调节HMGB1/TLR4信号通路对脂多糖诱导牙髓细胞凋亡的影响
- Author:
Yao LIN
1
;
Congna LIU
;
Shixia WANG
;
Zhiyong ZHANG
Author Information
- Keywords: HMGB1 protein; Toll-like receptor 4; acacia; lipopolysaccharides; pulpitis; apoptosis; cell proliferation; pulp cell
- From: Tianjin Medical Journal 2024;52(12):1238-1243
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the effect of acacetin(ACE)on the apoptosis of dental pulp cells induced by lipopolysaccharide(LPS)by regulating the high mobility histone 1(HMGB1)/Toll-like receptor 4(TLR4)signaling pathway.Methods Third generation dental pulp cells were isolated,cloned and purified from pulp of five patients with healthy teeth extracted from orthodontics and impaction.Cell morphology was observed and identification of stromal cell surface antigen1(STRO1)by immunofluorescence,and epidermal antigens CD90,CD105,CD34 and CD45 were identified by flow cytometry.MTT was applied to identify the effect of ACE on the activity of primary dental pulp cells(DPSCs).DPSCs were divided into the control group(0 μmol/L ACE culture),the LPS group(1 ng/L LPS treatment),the L-ACE group(added 20 μmol/L ACE on the basis of LPS group),the H-ACE group(added 40 μmol/L ACE on the basis of LPS group),the pcDNA-NC group(transfected pcDNA-NC plasmid on the basis of H-ACE group)and the HMGB group(transfected pcDNA-HMGB on the basis of H-ACE group).Cell proliferation was detected by CCK-8.Clone formation number was detected by clone formation assay.Western blot experiments were applied to detect the expression of HMGB1/TLR4 pathway,apoptosis and inflammation related proteins.ELISA assay was applied to detect levels of interleukin(IL-)-1β,IL-4 and tumor necrosis factor-α(TNF-α)in cell supernatant.Results DPSCs were successfully isolated and identified based on cell morphology,positive expression of STRO1,positive expression of CD90 and CD105,and negative identification of CD34 and CD45.Compared with the control group,cell proliferation activity,clonal cell count,B-cell lymphoma-2(Bcl-2)and IL-4 levels were decreased in the LPS group,while apoptosis rate,the pathway related proteins HMGB1,TLR4,apoptosis related proteins aspartate-specific cysteine protease(Caspase)-3,Caspase-7,Bcl-2 associated X protein(Bax),and inflammatory factors IL-1β,TNF-α were increased in the LPS group(P＜0.05).Compared with the LPS group,cell proliferation activity,Bcl-2 and IL-4 were increased successively in the L-ACE group and the H-ACE group,and apoptosis rate,pathway-related proteins HMGB1,TLR4,Caspase-3,Caspase-7 and Bax,and inflammatory factors IL-1β,TNF-α were reduced successively(P＜0.05).Overexpression of HMGB reversed the effects of H-ACE on cell proliferation,apoptosis,expression of related proteins and inflammatory factors(P＜0.05).Conclusion ACE inhibits LPS induced apoptosis of dental pulp cells,and it may be achieved by inhibiting the HMGB1/TLR4 signaling pathway.
