Baicalin activates autophagy,inhibits inflammation and promotes odontogenic/osteogenic differentiation of dental pulp stem cells
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1Department of General Dentistry,2Department of Endodontics,the Affiliated Stomatological Hospital of NanjingMedical University,State Key Laboratory Cultivation Base of Research,Prevention and Treatment for Oral Diseases,Jiangsu Province Engineering Research Center of Stomatological Translational Medicine,Nanjing 210029 ,China

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    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 -Ⅱ and 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.

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CUI Chuang, SUN Siyi, QIN Ziyi, SHA Ying, XU Hai, JIANG Fei, ZHANG Guangdong. Baicalin activates autophagy, inhibits inflammation and promotes odontogenic/osteogenic differentiation of dental pulp stem cells[J].,2025,(6):766-776.

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  • Received:March 03,2025
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  • Online: June 10,2025
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