正畸对牙龈炎大鼠牙周组织TLR4/NF-κB信号通路的调控作用研究

Regulatory effect of orthodontics on TLR4/NF-κB signaling pathway in periodontal tissue of gingivitis rats

目的探讨正畸对牙龈炎大鼠牙周组织中TLR4/NF-κB信号通路相关分子表达的影响,为临床牙龈炎、牙周炎患者的正畸矫治提供实验依据。方法将100只大鼠随机分为4组,包括牙龈炎组、正常加力50 g组、正常对照组、牙龈炎加力50 g组(实验组),每组25只。丝线结扎法构建牙龈炎模型,正畸镍钛螺旋拉簧构建加力模型,分别于第1、3、5、7、14天处死,取大鼠双侧实验区近中侧牙周组织。酶联免疫吸附试验(ELISA)检测大鼠牙周组织中白介素1β(IL-1β)、白介素6(IL-6)、肿瘤坏死因子(TNF-α)含量。Real-time PCR法、免疫组化测定大鼠牙周组织中TLR4、NF-κB p65的表达水平。结果实验组大鼠牙周组织中IL-1β、IL-6、TNF-α含量均明显高于对照组,差异具有统计学意义(P<0.05);Real-time PCR、免疫组化结果显示,实验组大鼠牙周组织中TLR4、NF-κB p65的表达均明显高于对照组,差异具有统计学意义(P<0.05)。TLR4、NF-κB p65含量变化呈正态分布,第5~7天为高峰期。结论正畸力可促进牙龈炎大鼠TLR4/NF-κB炎性信号通路的开放,从而加重正畸治疗中的牙周组织的局部炎性反应。

This study was designed to investigate the effect of orthodontics on TLR4/NF-κB signaling pathway-related molecules in gingivitis rats periodontal tissues,with a aim of providing experimental basis for gingivitis and periodontitis therapy.One hundred rats were randomly divided into four groups,including gingivitis group,normal force 50 g group,normal control group,gingivitis force 50 g group(experimental group),with 25 rats in each group.After gingivitis model was established by silk ligation,orthodontic Ni-Ti spiral tension spring was used to build force model.Rats were executed at the 1^st,3^rd,5^th,7^th,and 14^th days,and then the periodontal tissues in the mesal sides of the rat experimental area were taken.The concentration of interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor(TNF-α)in rat periodontal tissues were measured by enzyme-linked immunosorbent assay(ELISA),while the expression of TLR4 and NF-κB p65 in rat periodontal tissues was detected by real-time PCR and immunohistochemistry.Data showed that the concentration of IL-1β,IL-6 and TNF-αin rats’periodontal tissues with gingivitis in the experiment group was higher than those of the control group(P<0.05).The results of real-time PCR and immunohistochemistry showed that the expression of TLR4 and NF-κB p65 in the periodontal tissues of rats with gingivitis in the experiment group was also higher than those of the control group(P<0.05).The expression levels of TLR4,NF-κB p65 showed a normal distribution,and the expression peak appeared in day 5-7.Taken together,orthodontic force can promote the expression of TLR4/NF-κB inflammatory signaling pathway in gingivitis rats,therefore aggravating the local inflammatory responses in periodontal tissues during orthodontic treatment.