EZH2影响炎症状态下人牙周膜干细胞成骨分化能力的研究

EZH2 regulates LPS-induced HPDLSC osteogenesis

目的:探究人类同源基因2(EZH2)对脂多糖(Lipopolysaccharides,LPS)刺激下的牙周膜干细胞(periodontal ligament stem cells,PDLSCs)成骨分化能力的影响。方法:体外分离PDLSCs,使用Western Blot技术筛选出LPS体外刺激PDLSCs最佳浓度与时间模拟炎症环境,使用RNA沉默技术敲减EZH2基因,利用脂质体将siEZH2转染至PDLSCs细胞后使用Western blot检测EZH2的转染效率。使用脂质体转染技术敲低PDLSCs的EZH2基因后使用Western Blot检测EZH2的表达,Western Blot检测EZH2敲低对炎症状态下PDLSCs中成骨蛋白Runx2、碱性磷酸酶(Alkalinephosphatase,ALP)、骨钙素(osteocalcin,OCN)表达影响,使用茜素红染色检测PDLSCs成骨变化。结果:EZH2敲低后炎症状态下PDLSCs中骨形成标志物ALP、Runx2和OCN表达增强,茜素红染色矿化结节形成增加。结论:抑制EZH2可以促进LPS刺激所模拟炎症状态下PDLSCs成骨分化能力。

Objective:To explore the effect of EZH2 on the osteogenic differentiation of periodontal ligament stem cells(PDLSCs)stimulated by lipopolysaccharides(LPS).Methods:PDLSCs were isolated in vitro,and Western blot was used to screen out the optimal concentration and time of LPS to stimulate PDLSCs.Liposomal transfection technology was used to knock down the EZH2 gene of PDLSCs and Western blot was used to detect the expression of EZH2,and Western blot was used to detect Runx2,ALP,OCN in PDLSCs in inflammatory conditions.Alizarin red staining was used to detect the osteogenic changes of PDLSCs.Results:After EZH2 knockdown,the expression of ALP,Runx2 and OCN in PDLSCs increased in the inflammatory state,and the formation of mineralized nodules increased by Alizarin Red.Conclusions:Inhibition of EZH2 promoted osteogenic differentiation of PDLSCs under LPS-stimulated inflammatory conditions.