基于TLR4/NF-κB通路对固齿牙粉联合针刺疗法治疗牙周炎的机制研究

Mechanism of treating periodontitis with solid tooth powder combined with acupuncture based on the TLR4/NF-κB pathway

目的 研究固齿牙粉联合针刺疗法对实验性牙周炎大鼠的抗炎作用及作用机制,为中药治疗牙周炎提供实验依据。方法 以钢丝结扎大鼠上颌右侧第一磨牙、涂布牙龈卟啉单胞菌和饲喂高糖软饲料的方法建立实验性大鼠牙周炎模型。将60只大鼠随机分正常组、模型组、牙粉组、牙粉+针刺组、针刺组以及替硝唑组,每组10只。造模成功后,各药物组大鼠在牙齿上涂布相应药物,模型组大鼠涂布等体积生理盐水,每日3次,连续7 d。HE染色观察牙龈和牙槽骨组织形态学变化;RT-PCR法检测牙龈组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、Toll样受体4(TLR4)以及核因子κB p65(NF-κB p65)的m RNA表达水平;采用酶联免疫吸附法检测牙龈组织中TNF-α、IL-1β、IL-6的表达水平;采用Western Blot法测定大鼠牙龈组织中TLR4、NF-κB p65和p-NF-κB p65蛋白表达量。结果 与模型组比较,牙粉组以及牙粉+针刺组大鼠牙龈炎性浸润明显减少,牙龈组织TNF-α、IL-1β、IL-6含量以及TLR4、NF-κB p65和p-NF-κB p65蛋白表达水平均显著降低(P <0.05)。而牙粉组和牙粉+针刺组上述指标差异无统计学意义(P>0.05)。结论 固齿牙粉联合针刺疗法可促进牙周炎模型大鼠的牙周炎症状减轻,具有一定的抗炎作用,其机制可能与抑制TLR4/NF-κB信号通路、减少炎症反应有关。

Objective To study the anti-inflammatory effect and its related mechanisms of solid tooth powder combined with acupuncture in experimental periodontitis rats,and to provide experimental basis for the treatment of periodontitis with traditional Chinese medicine.Methods The periodontitis model rat was established by liqating the upper right first molar,coating porphyromonas gingivalis and feeding high-sugar soft feed.Sixty rats were randomly divided into normal group,model group,dental powder group,dental powder+acupuncture group,acupuncture group and tinidazole group,with 10 rats in each group.After successful modeling,rats in each drug group were coated with corresponding drugs on the teeth,and rats in the model group were coated with equal volume normal saline,3 times a day for 7 days.HE staining was used to observe the histomorphological changes of gum and alveolar bone.The mRNA expression levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),Toll-like receptor 4(TLR4)and nuclear factor-κB p65(NF-κB p65)were detected by RT-PCR.The expression levels of TNF-α,IL-1βand IL-6 in gingival tissues were detected by enzyme-linked immunosorbent assay.The expression levels of TLR4,NF-κB p65 and p-NF-κB p65 in gingival tissue of rats were determined by Western Blot.Results Compared with model group,qingival inflammatory infiltration in dental powder group and dental powder+acupuncture group were significantly reduced.The contents of TNF-α,IL-1βand IL-6 and the expression levels of TLR4,NF-κB p65 and p-NF-κB p65 in gum tissue of rats were significantly decreased(P<0.05).There was no significant difference in the above indexes between dental powder group and dental powder+acupuncture group(P>0.05).Conclusions Solid tooth powder combined with acupuncture can alleviate the symptoms of periodontitis in periodontitis model rats,and has certain anti-inflammatory effect,which may be related to inhibiting TLR4/NF-κB signaling pathway and reducing inflammatory response.