乳铁蛋白治疗牙周炎的炎症免疫机制研究

Study on Inflammatory Immune Mechanism of Lactoferrin in the Treatment of Periodontitis

目的 研究乳铁蛋白调节炎症免疫反应在治疗牙周炎中的作用及其机制。方法 取100只SD大鼠随机分成空白对照组,模型组,乳铁蛋白给药组低、中、高剂量组(1,2,3 g·kg^(-1)),甲硝唑阳性对照组(0.02 g·kg^(-1)),PDTC组(200 mg·kg^(-1)),乳铁蛋白+PDTC组(2 g·kg^(-1),200 mg·kg^(-1)),MCC950组(1 mg·kg^(-1)),乳铁蛋白+MCC950(2 g·kg^(-1),1 mg·kg^(-1)),每组10只。采用丝线结扎联合10%蔗糖饮水建立模型后开始给药,每天1次口腔给药,空白对照组和模型组口腔给药0.9%NaCl,连续给药1个月后处死各组大鼠。ELISA试剂盒检测IL-1β、IL-8、IL-10的含量;Western blotting检测TLR2-NF-κB通路和NLRP3炎症小体相关蛋白的表达。采用HE染色观察各组大鼠牙周组织的病理变化。结果 与模型组比较,乳铁蛋白各剂量组牙周炎症状得到明显改善,HE染色显示炎症细胞浸润减少,成纤维细胞增生活跃。TLR2、NF-κB、NLRP3、Caspase-1 p20和GSDMD-N蛋白表达降低,促炎因子IL-1β、IL-8的含量降低,抗炎因子IL-10的含量升高。结论 乳铁蛋白在治疗牙周炎中发挥的调节炎症免疫反应可能通过下调TLR2-NF-κB-NLRP3通路的蛋白表达,降低炎症反应的启动和炎症因子的释放,从而达到抗炎的目的。

OBJECTIVE To study the anti-inflammatory immune response effects of lactoferrin in the treatment of periodontitis and its mechanism.METHODS One hundred SD rats were randomly divided into blank control group,model group,lactoferrin administration group low,medium,high dose group(1,2,3 g·kg^(-1)),metronidazole positive control group(0.02 g·kg^(-1)),PDTC group(200 mg·kg^(-1)),lactoferrin+PDTC group(2 g·kg^(-1),200 mg·kg^(-1)),MCC950 group(1 mg·kg^(-1)) and lactoferrin+MCC950 group(2 g·kg^(-1),1 mg·kg^(-1)),10 rats in each group.Silk thread ligation combined with 10%sucrose drinking water was used to establish the model,and then the drug was administered orally once a day.The blank control group and the model group were administered orally with 0.9%NaCl.The rats in each group were sacrificed after one month of continuous administration.The contents of IL-1β,IL-8 and IL-10 were detected by ELISA kit,and the expressions of TLR2-NF-κB pathway and NLRP3 inflammasome related proteins were detected by Western blotting.HE staining was used to observe the pathological changes of the periodontal tissues of the rats in each group.RESULTS Compared with the model group,the symptoms of periodontitis in each dose group of lactoferrin were significantly improved.HE staining showed that the infiltration of inflammatory cells was reduced,and the proliferation of fibroblasts was active.The protein expressions of TLR2,NF-κB,NLRP3,Caspase-1 p20 and GSDMD-N decreased,the content of pro-inflammatory factor IL-8 and IL-1β decreased,and the content of anti-inflammatory factor IL-10 increased.CONCLUSION Lactoferrin may play a role in the regulation of inflammatory immune response in the treatment of periodontitis by down-regulating the protein expression of TLR2-NF-κB-NLRP3 pathway,reducing the initiation of inflammatory response and the release of inflammatory factors,so as to achieve the purpose of anti-inflammatory.