Abstract：Objective: To investigate the effect of eriodictyol on the osteogenic differentiation of human periodontal ligament stem cells (hPDLSC) induced by lipopolysaccharide (LPS) by regulating the Yes-associated protein (YAP)/transcriptional co-activator (TAZ) signaling pathway. Methods: The Periodontitis model of rats was established by the ligation method. After successful modeling, the rats were randomly divided into the model group and the eriodictyol group, with 6 rats in each group, and 6 normal rats as the control group. HPDLSC was cultured in vitro, induced with 10 μg/mL LPS, and treated with 0, 40, 80, 160, 320, and 480 μmol/L of eriodictyol, the alkaline phosphatase (ALP) activity of cells in each treatment group was detected with a kit to screen the optimal concentration of eriodictyol. hPDLSC were randomly divided into control group, LPS group, LPS+eriodictyol group, LPS+empty group, LPS+eriodictyol+YAP knockdown group, after inducing osteogenic differentiation and treated with LPS, eriodictyol, and plasmids. The pathological changes of periodontal tissue were observed by HE staining. Real-time fluorescence quantitative PCR and immunoblotting experiments were applied to detect the expression of YAP and TAZ in periodontal tissues and cells. ALP activity detection kit, ALP staining, and alizarin red staining were applied to detect osteogenic differentiation of cells. Enzyme linked immunosorbent assay (ELISA) was applied to detect the levels of inflammatory cytokines prostaglandin E2 (PGE2), interleukin (IL)-6 and IL-18 in periodontal tissues and cells. And real-time fluorescent quantitative PCR and Western blot were applied to detect the expression of osteogenic differentiation factors Runx2, OSX, and osteopontin (OPN) of cells. Results: Compared with the control group, the periodontal tissue of the model group showed obvious pathological damage, the serum levels of PGE2, IL-6 and IL-18 were increased, and the mRNA and protein expressions of YAP and TAZ were decreased (P<0.05). Compared with the model group, the pathological damage of periodontal tissue in the eriodictyol group was alleviated, the levels of serum PGE2, IL-6, and IL-18 were decreased, and the mRNA and protein expressions of YAP and TAZ in periodontal tissue were increased (P<0.05). The ALP activity in hPDLSC induced by LPS was increased after treatment with 40, 80, 160, 320, and 480 μmol/L eriodictyol (P<0.05), and the increasing effect was enhanced with the increase of the concentration of eriodictyol and reached a plateau at 320 μmol/L. Therefore, 320μmol/L sericol was selected for subsequent experiments. Compared with the control group, the expression of YAP and TAZ mRNA and protein, the relative proportion of ALP positivity, ALP activity, the relative proportion of mineralized nodules, the expression of Runx2 and OSX, OPN mRNA and protein in the LPS group decreased (P<0.05), the levels of PGE2, IL-6, and IL-18 increased (P<0.05). Compared with the LPS group, the expression of YAP and TAZ mRNA and protein, the relative proportion of ALP positivity, ALP activity, the relative proportion of mineralized nodules, the expression of Runx2 and OSX, OPN mRNA and protein in LPS+eriodictyol group increased (P<0.05), the levels of PGE2, IL-6, and IL-18 decreased (P<0.05); there was no obvious change in all indicators of cells in the LPS+empty group (P>0.05). Compared with the LPS+eriodictyol group, the expression of YAP and TAZ mRNA and protein, the relative proportion of ALP positivity, ALP activity, the relative proportion of mineralized nodules, the expression of Runx2 and OSX, OPN mRNA and protein in LPS+eriodictyol+YAP knockdown group decreased (P<0.05), the levels of PGE2, IL-6, and IL-18 increased (P<0.05). Conclusion: Eriodictyol can reduce the inflammatory damage of periodontitis rats, and inhibit LPS-induced hPDLSC inflammation by up-regulating the expression of YAP/TAZ pathway proteins, thereby promoting its osteogenic differentiation.
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2024, Vol. 30 
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