槲皮素对人口腔上皮细胞再生的生物学效应研究
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摘要
目的:研究槲皮素对人口腔角质细胞(human oral keratinocytes,HOKs)再生的生物学效应。方法:通过细胞计数试剂盒-8(cell counting kit-8,CCK-8)及迁移实验验证槲皮素对人牙龈角质细胞的增殖与迁移的药理作用。通过牙龈卟啉单胞菌脂多糖作为炎症刺激因子构建体外炎症模型,实时荧光定量聚合酶链反应(real time quantitative polymerase chain reaction,RT-qPCR)法检测转化生长因子(transforming growth factor,TGF)-β1和TGF-β3的表达及炎症因子白细胞介素(interleukin,IL)-1β和肿瘤坏死因子(tumor necrosis factor,TNF)-α的基因表达验证槲皮素的抗炎效果。酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测TGF-β1和TGF-β3的分泌水平。结果:20μmol/L槲皮素促进HOKs的增殖与迁移,并提高TGF-β3的mRNA水平与蛋白质表达,而50μmol/L槲皮素对TGF-β1的表达有促进作用。20、50μmol/L的槲皮素都有明显的抗炎作用。结论:20μmol/L槲皮素在体外实验中能够促进创面愈合并且促进HOKs再生。
Objective:To assess the potential of quercetin on gingival regeneration.Methods:Cell viability and proliferation capacity were tested by CCK-8 kit.Cell migration was monitored using Transwell chambers.An inflammatory situation was mimicked using Pg.LPS.RT-qPCR was performed to measure mRNA levels of IL-β1,TNF-α,TGF-β1,and TGF-β3.Protein levels of TGF-β1 and TGF-β3 were evaluated with ELISA.Results:20μmol/L quercetin enhanced the proliferation and migration of HOKs.Furthermore,20μmol/L quercetin increased both mRNA and protein levels of TGF-β3 under inflammatory conditions while 50μmol/L quercetin increased expression of TGF-β1.Expression of IL-β1 and TNF-αwere downregulated by quercetin treatment.Conclusion:Compared with untreated controls,the re-epithelialization of 20μmol/L quercetin-treated HOKs were highly rapid.
Objective:To assess the potential of quercetin on gingival regeneration.Methods:Cell viability and proliferation capacity were tested by CCK-8 kit.Cell migration was monitored using Transwell chambers.An inflammatory situation was mimicked using Pg.LPS.RT-qPCR was performed to measure mRNA levels of IL-β1,TNF-α,TGF-β1,and TGF-β3.Protein levels of TGF-β1 and TGF-β3 were evaluated with ELISA.Results:20μmol/L quercetin enhanced the proliferation and migration of HOKs.Furthermore,20μmol/L quercetin increased both mRNA and protein levels of TGF-β3 under inflammatory conditions while 50μmol/L quercetin increased expression of TGF-β1.Expression of IL-β1 and TNF-αwere downregulated by quercetin treatment.Conclusion:Compared with untreated controls,the re-epithelialization of 20μmol/L quercetin-treated HOKs were highly rapid.
引文
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[17]闫福华,李丽丽.牙周再生治疗研究进展[J].口腔医学研究,2018,34(3)∶217-222
[2]Smith PC,Cceres M,Martínez C,et al.Gingival wound healing:an essential response disturbed by aging[J].J Dent Res,2015,94(3)∶395-402
[3]Hasan A,Palmer RM.A clinical guide to periodontology:Pathology of periodontal disease[J].Br Dent J,2014,216(8)∶457-461
[4]Polimeni G,Xiropaidis AV,Wikesj9UM.Biology and principles of periodontal wound healing/regeneration[J].Periodontol 2000,2006,41(1)∶30-47
[5]Nieto MA,Huang RY,Jackson RA,et al.EMT:2016[J].Cell,2016,166(1)∶21-45
[6]Shimoe M,Yamamoto T,Shiomi N,et al.Overexpression of Smad2inhibits proliferation of gingival epithelial cells[J].J Periodontal Res,2014,49(3)∶290-298
[7]Chang Z,Kishimoto Y,Hasan A,et al.TGF-β3modulates the inflammatory environment and reduces scar formation following vocal fold mucosal injury in rats[J].Dis Model Mech,2014,7(1)∶83-91
[8]Eslami A,GallantBehm CL,Hart DA,et al.Expression of integrinαvβ6and TGF-βin scarless vs scar-forming wound healing[J].J Histochem Cytochem,2009,57(6)∶543-557
[9]Hatahet T,Morille M,Hommoss A,et al.Quercetin topical application,from conventional dosage forms to nanodosage forms[J].Eur J Pharm Biopharm,2016,108∶41-53
[10]Pietta PG.Flavonoids as antioxidants[J].J Nat Prod,2000,63(7)∶1035-1042
[11]Gómez-Florit M,Monjo M,Ramis JM.Quercitrin for periodontal regeneration:effects on human gingival fibroblasts and mesenchymal stem cells[J].Sci Rep,2015,5∶16593
[12]Gomez-Florit M,Pacha-Olivenza MA,Fernandez-Calderon MC.Quercitrin-nanocoated titanium surfaces favour gingival cells against oral bacteria[J].Sci Rep,2016,6∶22444
[13]Larjava H,Hkkinen L,Koivisto L.Re-Epithelialization of Wounds[M].Oral Wound Healing.John Wiley&Sons,Ltd.2011.81-123
[14]SatuéM,Arriero MM,Monjo M,et al.Quercitrin and taxifolin stimulate osteoblast differentiation in MC3T3-E1cells and inhibit osteoclastogenesis in RAW 264.7cells[J].Biochem Pharmacol,2013,86(10)∶1476-1486
[15]Vicentini FT,He T,Shao Y,et al.Quercetin inhibits UV irradiation-induced inflammatory cytokine production in primary human keratinocytes by suppressing NF-kappaB pathway[J].J Dermatol Sci,2011,61(3)∶162-168
[16]Caddeo C,Sales OD,Valenti D,et al.Inhibition of skin inflammation in mice by diclofenac in vesicular carriers:liposomes,ethosomes and PEVs[J].Int J Pharm,2013,443(1-2)∶128-136
[17]闫福华,李丽丽.牙周再生治疗研究进展[J].口腔医学研究,2018,34(3)∶217-222