南京医科大学学报（自然科学版）2025，Vol.45Issue(6)：766-776,11.DOI:10.7655/NYDXBNSN250241
黄芩苷激活自噬抑制炎症促进牙髓干细胞成牙/骨分化
Baicalin activates autophagy,inhibits inflammation and promotes odontogenic/osteogenic differentiation of dental pulp stem cells
摘要
Abstract
Objective:This study aims to verify the anti-inflammatory effect of baicalin(BA)in an inflammatory environment and explore its influence on the odontogenic/osteogenic differentiation of human dental pulp stem cells(hDPSC),providing a reference for the vital pulp therapy of pulpitis.Methods:Select the early inflammatory pulp tissue of the human teeth for making frozen sections and conducting HE staining,interleukin(IL)-1β immunohistochemical staining,as well as IL-1β and inducible nitric oxide synthase(iNOS)immunofluorescence staining.Human monocyte leukemia cells(THP-1)were differentiated into macrophages and polarized to the M1 subtype under the induction of lipopolysaccharide(LPS)and interferon-γ(IFN-γ).Then,cells were treated with BA(50 μmol/L),the expression levels of iNOS,IL-1β,and IL-8 were analyzed using immunofluorescence,real-time quantitative PCR(RT-qPCR),and Western blot.Additionally,the expression changes of LC3B,Beclin-1,and P62 were assessed by Western blot to monitor autophagic flux alterations in macrophages.To further investigate the relationship between inflammation inhibition and autophagy,we employed the autophagy inhibitor 3-MA to block autophagic flux and re-evaluated the expression changes of IL-1β and IL-8.Supernatants from macrophages cultured under various conditions were collected,and the conditioned medium was prepared and added to mineralized hDPSC.The odontogenic/osteogenic differentiation ability of hDPSC in an inflammatory environment was analyzed by alkaline phosphatase ALP staining,alizarin reds(ARS)staining,RT-PCR,and Western blot.Results:The expression of IL-1β and iNOS around the inflammatory area of early pulpitis tissue was significantly increased.After induced by LPS and IFN-γ,macrophages were polarized from M0 to M1,and the expressions of iNOS,IL-1β and IL-8 significantly increased.After co-incubation with 50 μmol/L BA for 24 hours,the polarization degree of M1 macrophages significantly decreased,the mRNA levels of IL-1β and IL-8 significantly decreased compared with the M1 group.Compared with the M1 group,the expressions of LC3B-Ⅱand Beclin-1 increased after the addition of BA,while the expression of P62 was inhibited.After 3-MA blocked autophagy,the mRNA levels of IL-1β and IL-8 significantly increased.After adding the supernatant of M1 macrophages to hDPSC and inducing mineralization for 7～21 days,the ALP activity of hDPSC decreased,the calcium salt deposition significantly reduced,and the expressions of ALP,DSPP,RUNX2,OPN and COL-1 significantly decreased.When adding the supernatant of M1 macrophages treated with BA,the ALP activity of hDPSC significantly increased,the calcium salt deposition significantly increased,and the expressions of ALP,DSPP,RUNX2,OPN and COL-1 significantly increased.Conclusion:BA can inhibit inflammatory responses by activating autophagy,thereby enabling hDPSC to function in an inflammatory microenvironment.关键词
黄芩苷/M1巨噬细胞/THP-1/成牙/骨分化/牙髓干细胞Key words
baicalin/M1 macrophages/THP-1/odontogenic/osteogenic differentiation/dental pulp stem cell分类
口腔医学引用本文复制引用
崔闯,孙思怡,秦梓一,沙颖,徐海,江飞,张光东..黄芩苷激活自噬抑制炎症促进牙髓干细胞成牙/骨分化[J].南京医科大学学报（自然科学版）,2025,45(6):766-776,11.基金项目
国家自然科学基金(82270955) （82270955）
南京市卫生科技发展专项资金(YKK24293) （YKK24293）
