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Abstract （Department of Stomatology, Hebei Petro China Central Hospital, Langfang 065000, China）
Abstract: To investigate the effects of luteic acid on inflammation and apoptosis of human periodontal stem cells induced by lipopolysaccharide and the regulation of Wnt/β-catenin signaling pathway, hPDLSCs were cultured in vitro, which was divided into two steps: pre-experiment and formal experiment. The pre-experiment was divided into control group (no intervention), lipopolysaccharide group (10 μg/mL lipopolysaccharide), low/medium/high concentration luboflavic acid group (10 μg/mL lipopolysaccharide +0.5, 1.0, 2.0 μmol/L luboflavic acid). The formal experiment was divided into control group, lipopolypaccharide group, luteic acid group (10 μg/mL lipopolypaccharide +2.0 μmol/L luteic acid), inhibitor group (10 μg/mL lipopolypaccharide +2 μmol/L Wnt/β-catenin signaling pathway inhibitor XAV939), luteic acid + inhibitor group (10 μg/mL lipopolysaccharide +2.0 μmol/L luteic acid +2 μmol/L XAV939) and luteic acid + activator group (10 μg/mL lipopolysaccharide +2.0 μmol/L luteic acid +20 μmol/L Wnt/β-catenin signaling pathway activator SKL2001), they were treated for 24 h. Cell viability was measured by CCK-8. The expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were detected by enzyme-linked immunosorbent assay (ELISA). The apoptosis rate was detected by Hoechst 33258 staining. The expression levels of B lymphoblastoma-2 (Bcl-2), cysteine aspartate protease-3 (Caspase-3) and Wnt/β-catenin signaling pathway were detected by Western blot (Wb). In the preliminary experiment, the cell viability of LPS group decreased compared with control group (P<0.05), and the expression levels of inflammatory factors TNF-α, IL-1β and IL-6 increased (P<0.05). Compared with LPS group, the cell viability of high-concentration luteic acid group increased (P<0.05), and the levels of inflammatory factors TNF-α, IL-1β and IL-6 decreased (P<0.05). Therefore, the high-concentration luteic acid group was selected as the optimal concentration for formal experiment. In the formal experiment, the cell viability and anti-apoptotic Bcl-2 protein expression levels in LPS group decreased compared with those in control group (P<0.05), while the expression levels of inflammatory factors TNF-α, IL-1β and IL-6, apoptosis rate and the expression levels of Caspase-3, Wnt and β-catenin proteins increased (P<0.05). Compared with LPS group, the cell viability and anti-apoptotic Bcl-2 protein expression levels in luteic acid group and inhibitor group increased (P<0.05), while the expression levels of inflammatory factors TNF-α, IL-1β, IL-6, cell apoptosis rate and the expression levels of Caspase-3, Wnt and β-catenin proteins decreased (P<0.05). Compared with the luteic acid group, the change trend of the above indexes in the luteic acid + inhibitor group was more significant (P<0.05), and the change trend of these indexes in the luteic acid + activator group was significantly reversed (P<0.05). Luteic acid may promote the cellular activity of human hPDLSCs induced by lipopolysaccharide and inhibit their apoptosis and inflammation by inhibiting the signal transduction of Wnt/β-catenin signaling pathway. This study can provide a new theoretical reference for the treatment of periodontitis.
Key words: periodontitis; gambogic acid; lipopolysaccharide; periodontal membrane stem cells; Wnt/β-catenin signaling pathway
（Acta Laser Biology Sinica, 2024, 33(6): 567-576）
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. [J]. journal1, 2024, 33(6): 1-6. |
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