烟草提取物对牙周炎小鼠牙周组织TLR4炎症通路的影响

Effects of Tobacco Extract on TLR4 Inflammatory Pathway in Periodontal Tissues of Mice with Periodontitis

目的:探究烟草提取物对牙周炎小鼠牙周组织中Toll样受体4(TLR4)炎症通路的影响。方法:将小鼠随机分为对照租、模型组、烟草提取物低、中、高剂量组。Micro-CT扫描观察牙周骨骼变化;酶联免疫法(Elisa)检测血清中肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)水平;HE染色检测牙周组织病理形态学变化;实时荧光定量PCR(qRT-PCR)、免疫印迹(WB)、免疫组化法检测牙周组织中TLR4、肿瘤坏死因子受体相关因子6(TRAF6)、核因子κB(NF-κB)表达。结果:模型组骨体积比、骨表面积比、骨小梁厚度均显著降低,骨小梁间隙增大,血清中TNF-α、IL-1β、IL-6水平、牙周组织中TLR4、TRAF6、NF-κB表达均显著升高(P<0.05)。与模型组相比,烟草提取物各组骨体积比、骨表面积比、骨小梁厚度降低,骨小梁间隙增大,血清TNF-α、IL-1β、IL-6水平、牙周组织中TLR4、TRAF6、NF-κB表达均显著升高(P<0.05)。结论:烟草提取物可加重牙周炎小鼠牙周组织炎症反应,其机制可能与上调牙周组织中TLR4信号通路有关。

Objective: To explore the effects of tobacco extract on Toll-like receptor 4 (TLR4) inflammatory pathway in periodontal tissues of mice with periodontitis.Methods: Rats were randomly divided into control group,model group,low,medium,and high dose groups.Micro-CT scan was used to observe periodontal bone changes.The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6 (IL-6) in serum were detected by enzyme-linked immunoassay (Elisa).HE staining was used to detect the pathomorphological changes of periodontal tissues.The expressions of TLR4,tumor necrosis factor receptor related factor 6 (TRAF6),and nuclear factor κB (NF-κB) in periodontal tissues were detected by real-time fluorescence quantitative PCR (qRT-PCR),immunoblotting (WB),and immunohistochemistry.Results: Bone volume ratio,bone surface area ratio,and trabecular thickness in model group were significantly decreased,trabecular space was increased,serum levels of TNF-α,IL-1β,IL-6,expression of TLR4,TRAF6,and NF-κB in periodontal tissue were significantly increased ( P < 0.05 ).Compared with the model group,the bone volume ratio,bone surface area ratio,trabecular thickness,trabecular space,serum levels of TNF-α,IL-1β,IL-6,expression of TLR4,TRAF6 and NF-κB in periodontal tissues of the tobacco extract groups were significantly increased ( P <0.05).Conclusion: Tobacco extract can aggravate periodontal tissue inflammatory reaction in mice with periodontitis,which may be related to up-regulation of TLR4 signaling pathway in periodontal tissue.