Abstract:

Objective: To explore the mechanism of Baicalin （BA） in promoting odontogenic/osteo differentiation and anti-inflammation in inflammatory pulp. Methods: Dental pulp stem cells (DPSCs) were isolated and extracted by enzyme digestion method.CCK-8 was used to detect the effect of BA on the activity of DPSCs and mononuclear macrophages cells (THP-1). ALP activity detection and staining were used to select the optimal concentration of BA to promote the expression of ALP in DPSCs. LPS was used to construct an inflammatory microenvironment in vitro. Real-time quantitative PCR and Western blot were used to detect the changes in the expression levels of proteins and genes related to odontogenic/osteo differentiation of DPSCs induced by BA. The expression levels of autophagy associated ubiquitin-binding protein (p62) and LC3Ⅱ/Ⅰ were detected by immunofluorescence staining and Western bolt. THP-1 was induced into inflammatory M1 state, and the effects of BA on the expression of (IL-1β, TNF-α and iNOS were detected by qRT-PCR. Results: BA at the concentration of ≤30 μmol/L did not significantly inhibit the proliferation of DPSCs (P＞0.05). There was no significant difference in THP-1 among the groups exposed to BA at the concentration of 0-100  μmol/L (P＞0.05), and the proliferation of THP-1 cells was not significantly inhibited. The ALP activity of LPS-stimulated DPSCs was significantly increased when treated with 10 μmol/L BA (P＜0.01). The expressions of DSP, COL-I, RUNX2, OSX, and OCN were up-regulated (P＜0.05),while autophagy related protein p62 was down-regulated and LC3 Ⅱ/Ⅰ expression was up-regulated (P＜0.01). The expressions of IL-1β, TNF-α and iNOS in THP-1 were increased by LPS (P＜0.01), and these inflammatory indexes were down-regulated by 10 μmol/L BA (P＜0.05). Conclusions: BA may play an anti-inflammatory role in inflammatory pulp cells by regulating autophagy and promoting odontogenic/osteo differentiation.