红景天苷对LPS刺激人牙周膜细胞增殖和分泌表达的影响

目的研究红景天苷(salidroside,SAL)对大肠杆菌脂多糖(lipopolysaccharide,LPS)刺激后体外培养人牙周膜细胞(human periodontal ligament cells,h PDLCs)的增殖及对分泌表达Toll样受体(Toll-like receptor 4,TLR 4)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、骨保护素(osteoprotegerin,OPG)及核因子-κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)的影响,为红景天苷在牙周炎治疗方面提供初步的实验依据。方法组织块法原代培养正常h PDLCs;以2 mg/L LPS刺激第4代细胞,加入不同浓度的红景天苷作用12、24、48 h;采用MTT方法检测细胞的增殖;采用Western blot、ELISA、qRT-PCR等方法检侧TLR-4、TNF-α、IL-1β、IL-6、OPG及RANKL的表达水平,并进行统计学分析。结果 MTT结果显示低浓度的红景天苷(≤10μmol/L)均能促进细胞增殖,并存在浓度依赖性,其中0.5μmol/L对h PDLCs增殖作用最明显(P<0.05),高浓度的红景天苷(>10μmol/L)不能促进牙周膜细胞增殖;Western blot、ELISA及qRT-PCR结果显示:0.5μmol/L红景天苷可使LPS刺激的h PDLCs的TLR4、TNF-α、IL-1β、IL-6及RANK表达水平随时间延长而逐渐降低,其中48 h降低的最为明显,存在显著性差异(P<0.05);另外,LPS刺激对OPG的表达几乎没有影响,而0.5μmol/L红景天苷作用可以显著上调OPG的表达水平。结论红景天苷可减轻LPS刺激后h PDLCs的细胞损伤,其机制可能通过调节与TLR4相关信号通路,降低炎症因子TNF-α、IL-1β、IL-6及与骨吸收相关因子RANKL、OPG的表达,从而抑制炎症反应和骨吸收。

Effect of salidroside on the proliferation and secretion of human periodontal ligament cells stimulated by lipopolysaccharide

Objective To investigate the effect of salidroside ( SAL ) on the proliferation and secretion of Toll?like receptor 4 ( TRL4) , tumor necrosis factor?α( TNF?α) , interleukin?1β( IL?1β) , interleukin?6 ( IL?6) , osteoprotegerin ( OPG) and receptor ac?tivator of NF?κB ligand ( RANKL) in human periodontal ligament cells ( hPDLCs) treated by lipopolysaccharide ( LPS) and to provide preliminary experimental basis for treatment of periodontitis by salidroside. Methods hPDLCs were cultured by tissue cultivation in vitro and treated with 2 mg/L LPS and various concentrations of salidroside for 12, 24 and 48 h. Cell proliferation of hPDLCs was de?tected by MTT assay. The expression levels of TRL4 TNF?α, IL?1β, IL?6, RANKL and OPG in hPDLCs were examined by Western blot, ELISA or qRT?PCR analysis. The data were processed by statistical analysis. Results Salidroside at low concentrations (≤10μmol/L) significantly promoted hPDLCs proliferation in dose?dependent manner; 0. 5 μmol/L of salidroside had the greatest effect whereas high concentrations of salidroside (>10 μmol/L) had no obvious effect on cell proliferation of hPDLCs; The expression of TLR4, TNF?α, IL?1β, IL?6 and RANKL was significantly increased by LPS stimulation. After treatment with 0.5 μmol/L of salidro?side, the expression levels of TLR4, TNF?α, IL?1β, IL?6 and RANKL were significantly inhibited and the obvious effect occurred at 48 h ( P<0.05);In addition, the expression of OPG was nearly unchanged by LPS, but significantly upregulated by 0.5μmol/L of sal?idroside. Conclusion Salidroside could attenuate LPS?induced cell injury in hPDLCs through inhibiting TLR4 signaling pathways and the production of inflammation factors including TNF?α, IL?1β, IL?6 and RANKL and promoting OPG expression which thereby inhibi?ted inflammatory reaction and bone resorption.