烟草提取物对牙周炎小鼠牙周组织TLR4炎症通路的影响
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摘要
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.
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.
引文
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[12] 张任飞,杨泓,郭立新.Nod样受体蛋白3和白细胞介素1β在2型糖尿病合并慢性牙周炎中的作用[J].中国糖尿病杂志,2017,9(8):509-514.
[13] 黄敏福.牙周治疗对牙周炎伴冠心病患者血清C反应蛋白、炎症因子及血脂水平的影响[J].中外医学研究,2017,15(1):20-21.
[14] 陈庆勇,李霞,张芳,等.TRAF6沉默对LPS刺激牙周膜成纤维细胞增殖能力的影响[J].口腔医学研究,2017,33(11):1181-1184.
[15] Sun Y,Huang J,Song K.BET protein inhibition mitigates acute myocardial infarction damage in rats via the TLR4/TRAF6/NF-κB pathway [J].Exp Ther Med,2015,10(6):2319-2324.
[16] 韩倩倩,戚威娟,刘钊,等.Toll样受体4抑制剂对糖尿病小鼠体内肿瘤坏死因子α及精氨酸酶1表达的影响[J].中华口腔医学研究杂志(电子版),2017,11(3):142-148.
[2] 尹伟,王晨辰,刘向辉.不同因素对牙种植体稳定性的影响[J].安徽医学,2016,37(2):249-251.
[3] 邵珺雅,汪建中.烟草对人牙龈成纤维细胞影响的研究进展[J].南昌大学学报(医学版),2016,56(4):95-97.
[4] 瞿凤环,邓婧,潘克清,等.局部注射LPS对慢性牙周炎模型大鼠牙髓、修复性牙本质形成的影响[J].山东医药,2012,52(18):36-37.
[5] Chapman KR,Mannino DM,Soriano JB,et al.Epidemiology andcosts of chronic obstructive pulmonary disease [J].Eur Respir,2006,27(8) :188-207.
[6] 瞿凤环,邓婧,潘克清,等.局部注射LPS对慢性牙周炎模型大鼠牙髓、修复性牙本质形成的影响[J].山东医药,2012,52(18):36-37.
[7] 王中秀,杨岚,谭静怡,等.辅助性T细胞1和17细胞特征性分泌因子在大鼠实验性牙周炎模型中的表达及意义[J].中华口腔医学杂志,2017,52(12):194-196.
[8] Lee SY,Nho TH,Choi BD,et al.Secretory leukocyte protease inhibitor reduces inflammation and alveolar bone resorption in LPS-induced periodontitis in rats and in MC3T3-E1 preosteoblasts [J].Animal Cells Systems,2016,20(6):1-9.
[9] Kubota M,Yanagita M,Mori K,et al.The effects of cigarette smoke condensate and nicotine on periodontal tissue in a periodontitis model mouse[J].PLoS One,2016,11(5):e0155594.
[10] 周芹,孙睿男,朱震坤.热休克蛋白27在烟草烟雾提取物介导的牙龈成纤维细胞损伤中的表达变化[J].华西口腔医学杂志,2016,34(1):17-22.
[11] 林梅,张冬雪,刘志强,等.慢性牙周炎患者血清IL-33、TNF-α、IL-6的检测及意义[J].首都医科大学学报,2016,37(3):255-259.
[12] 张任飞,杨泓,郭立新.Nod样受体蛋白3和白细胞介素1β在2型糖尿病合并慢性牙周炎中的作用[J].中国糖尿病杂志,2017,9(8):509-514.
[13] 黄敏福.牙周治疗对牙周炎伴冠心病患者血清C反应蛋白、炎症因子及血脂水平的影响[J].中外医学研究,2017,15(1):20-21.
[14] 陈庆勇,李霞,张芳,等.TRAF6沉默对LPS刺激牙周膜成纤维细胞增殖能力的影响[J].口腔医学研究,2017,33(11):1181-1184.
[15] Sun Y,Huang J,Song K.BET protein inhibition mitigates acute myocardial infarction damage in rats via the TLR4/TRAF6/NF-κB pathway [J].Exp Ther Med,2015,10(6):2319-2324.
[16] 韩倩倩,戚威娟,刘钊,等.Toll样受体4抑制剂对糖尿病小鼠体内肿瘤坏死因子α及精氨酸酶1表达的影响[J].中华口腔医学研究杂志(电子版),2017,11(3):142-148.