柚皮素通过基质细胞衍生因子1/趋化因子受体4信号轴对脂多糖作用下人牙周膜干细胞抗炎、成血管和成骨分化能力的影响

Effect of naringenin on the anti-inflammatory,vascularization,and osteogenesis differentiation of human periodontal ligament stem cells via the stromal cell-derived factor 1/C-X-C motif chemokine receptor 4 signaling axis stimulated by lipopolysaccharide

目的探究柚皮素(Nar)对脂多糖(LPS)刺激下的人牙周膜干细胞(hPDLSCs)抗炎、成血管和成骨能力的影响及其机制。方法采用细胞计数试剂盒(CCK-8)、划痕试验和Transwell实验研究hPDLSCs的增殖和迁移能力。采用碱性磷酸酶(ALP)染色、茜素红染色、管腔形成实验、酶联免疫吸附实验(ELISA)、实时荧光定量逆转录聚合酶链反应(qRT-PCR)和蛋白印迹实验(Western blot)检测hPDLSCs抗炎、成血管和成骨分化能力。结果10μmol/L Nar可减轻10μg/mL LPS刺激的hPDLSCs炎症反应,促进其增殖、迁移和成血管分化,0.1μmol/L Nar可有效恢复10μg/mL LPS刺激的hPDLSCs成骨能力。加入CXCR4抑制剂AMD3100后,Nar促进抗炎和成骨分化的作用降低,炎性hPDLSCs成血管分化升高。结论Nar促进了hPDLSCs抗炎、成血管和成骨分化,该作用与基质细胞衍生因子1/趋化因子受体4信号轴有关。

Objective This study aimed to investigate how naringenin(Nar)affected the anti-inflammatory,vascularization,and osteogenesis differentiation of human periodontal ligament stem cells(hPDLSCs)stimulated by lipopolysaccharide(LPS)and to preliminarily explore the underlying mechanism.Methods Cell-counting kit-8(CCK8),cell scratch test,and Transwell assay were used to investigate the proliferation and migratory capabilities of hPDLSCs.Alkaline phosphatase(ALP)staining,alizarin red staining,lumen-formation assay,enzyme-linked immunosorbent assay,quantitative timed polymerase chain reaction,and Western blot were used to measure the expression of osteopontin(OPN),Runt-related transcription factor 2(RUNX2),vascular endothlial growth factor(VEGF),basic fibroblast growth factor(bFGF),von Willebrand factor(vWF),tumor necrosis factor-α(TNF-α),and interleukin(IL)-6.Results We observed that 10μmol/L Nar could attenuate the inflammatory response of hPDLSCs stimulated by 10μg/mL LPS and promoted their proliferation,migration,and vascularization differentiation.Furthermore,0.1μmol/L Nar could effectively restore the osteogenic differentiation of inflammatory hPDLSCs.The effects of Nar’s anti-inflammatory and promotion of osteogenic differentiation significantly decreased and inflammatory vascularization differentiation increased after adding AMD3100(a specific CXCR4 inhibitor).Conclusion Nar demonstrated the ability to promote the anti-inflammatory,vascularization,and osteogenic effects of hPDLSCs stimulated by LPS,and the ability was associated with the stromal cell-derived factor/C-X-C motif chemokine receptor 4 signaling axis.