地塞米松对脂多糖介导下人牙周膜干细胞骨向分化过程中的调控作用  

Effect of dexamethasone on lipopolysaccharide mediated osteogenic differentiation of human periodontal ligament stem cells

作　　者：何晓光[1] 赵君[1] 李娜[1] HE Xiaoguang;ZHAO Jun;LI Na(Department of Pharmacy,Hefei Stomatological Hospital,Hefei 230001,China)

机构地区：[1]合肥市口腔医院安徽医科大学合肥口腔临床学院药剂科,安徽合肥230001

出　　处：《皖南医学院学报》2022年第2期166-169,共4页Journal of Wannan Medical College

基　　金：合肥市自主创新政策“借转补”资金项目(J2019Y03)。

摘　　要：目的:探讨地塞米松(Dex)对脂多糖(LPS)引起的炎症状态下人牙周膜干细胞骨向分化过程的影响及其可能的分子机制。方法:体外组织块法培养人牙周膜干细胞,并进行干细胞功能鉴定,取生长状况良好的第3代牙周膜干细胞,分为4组,正常对照组、1×10^(-8)mol/L Dex组、10μg/mL LPS组、1×10^(-8)mol/L Dex+10μg/mL LPS组。培养7 d后Elisa法检测炎症因子IL-6的表达情况;qRT-PCR检测骨相关基因ALP、RUNX-2、BSP表达水平;培养21 d后茜素红染色观察钙化结节形成情况;Western blot检测核蛋白中p65表达情况。结果:成功分离培养的人牙周膜干细胞,具有克隆形成能力以及多向分化的潜能;Elisa检测结果显示,LPS组IL-6的表达水平升高(P<0.05),Dex+LPS组IL-6的表达水平较LPS组降低(P<0.05);qRT-PCR结果显示Dex组骨相关基因ALP、RUNX-2、BSP表达水平上调(P<0.05),Dex+LPS组ALP、RUNX-2、OPN表达水平较Dex组下调(P<0.05);茜素红染色结果显示,Dex组钙化结节最多,Dex+LPS组次之,LPS组与空白对照组无钙化结节形成;Western blot检测结果显示,核蛋白中LPS组p65表达量较高(P<0.05),Dex+LPS组p65表达较LPS组降低(P<0.05)。结论:Dex具有促进牙周膜干细胞成骨分化及抗炎的作用,其可能是由于抑制了NF-κB信号通路的关键分子p65入核降低了炎症反应的发生。Objective:To investigate the effect of dexamethasone(Dex)on osteogenic differentiation of human periodontal ligament stem cells in lipopolysaccharide(LPS)-induced inflammation and its possible molecular mechanism.Methods:Human periodontal ligament stem cells were cultured with in vitro technique,and the function of stem cells was identified.The third generation of periodontal ligament stem cells were obtained,and divided into blank control group,1×10^(-8) mol/L Dex group,10μg/mL LPS group and 1×10^(-8) mol/L Dex+10μg/mL LPS group.Seven days after culturing,IL-6 expression was detected by ELISA,and expression levels of ALP,Runx-2 and BSP were measured by qRT-PCR.By day 21 after culturing,alizarin red staining was performed to observe the formation of calcified nodules,and Western blot was used to assay the expression of p65.Results:Human periodontal ligament stem cells were successfully isolated and cultured,which had the ability of clone formation and multi-directional differentiation.ELISA results showed that IL-6 expression in LPS group was significantly higher than that in Dex+LPS group(P<0.05).qRT-PCR demonstrated that the expression levels of ALP,Runx-2 and BSP were up-regulated in Dex group(P<0.05),whereas down regulated in Dex+LPS group compared to Dex group(P<0.05).Alizarin red staining exhibited that calcified nodules were most found in Dex group,followed by Dex+LPS group,yet no calcified nodules were seen in LPS group and blank control group.Western blot assay indicated that p65 expression in LPS group was higher than that in Dex+LPS group(P<0.05).Conclusion:Dexamethasone can promote the osteogenic differentiation of and anti-inflammatory effect on the periodontal ligament stem cells.The potential mechanism may be associated with inhibition of p65,the key molecule of NF-κB signaling pathway,into the nucleus and the reduction of inflammatory reaction.

关 键 词：地塞米松 脂多糖 牙周膜干细胞 骨向分化 

分 类 号：R781.4[医药卫生—口腔医学]