TLR4对人牙周膜细胞分泌IL-1β、IL-6和TNF-α的影响

Effect of TLR4 on production of IL-1β,IL-6 and TNF-α in human periodontal ligament cells

目的观察在内毒素脂多糖(LPS)刺激下,Toll样受体4(TLR4)对人牙周膜成纤维细胞(HPDLC)分泌白细胞介素(IL)中的IL-1β、IL-6和肿瘤坏死因子α(TNF-α)的影响。方法选用100ng/mL、1μg/mL和10μg/mL大肠杆菌LPS分别刺激HPDLC,双抗体夹心法检测刺激后6、12、24、48h时,HPDLC分泌IL-1β、IL-6和TNF-α的水平。运用不同滴度anti-TLR4单克隆抗体预先处理HPDLC,观察1μg/mLLPS刺激下其分泌IL-1β、IL-6和TNF-α的水平变化。结果LPS刺激HPDLC6h后,即可检测到IL-1β、IL-6和TNF-α,24h达到顶峰,然后逐渐下降,各LPS浓度组规律基本一致;anti-TLR4单克隆抗体预先处理的HPDLC,在1μg/mLLPS刺激下,其产生IL-1β、IL-6和TNF-α的水平明显下降(P<0.05),且与anti-TLR4单克隆抗体滴度呈量效关系。结论TLR4参与了HPDLC的炎症反应过程;anti-TLR4单克隆抗体能有效抑制LPS刺激后HPDLC分泌IL-1β、IL-6和TNF-α的能力。

Objective To observe the effect of Toll-like receptor 4 （TLR4） on production of interleukin （IL）-1β IL-6 and tumor necrosis factor-α （TNF-α） in human periodontal ligament cells （HPDLC） stimulated by lipopolysaccharide（LPS）. Methods The level of IL-1β, IL-6 and TNF-α in HPDLC stimulated by 100 ng/mL,1 μg/mL and 10 μg/mL LPS after 6, 12, 24 and 48 h were detected by double antibody sandwich ELISA. The level of IL-1β IL-6 and TNF-α in HPDLC stimulated by 1 μg/mL LPS after pretreatment with anti-TLR4 monoclonal antibody were observed. Results Productions of IL-1β IL-6 and TNF-α in the cultured medium were detected 6 h after stimulation with LPS, and the levels of these cytokines were highest at 24 h, and then were gradually decreased. After pretreatment with anti-TLR4 monoclonal antibody, the levels of these cytokines were significantly decreased （ P 〈 0.05） , and the effects of TLR4 on these cytokines were in a dose-dependent manner. Conclusion TLR4 plays a role in the inflammatory reaction of HPDLC. Anti- TLR4 monoclonal antibody can effectively suppress the productions of IL-1β IL-6 and TNF-α in HPDLC stimulated by LPS.