罗伊氏乳杆菌提取物对牙周炎症微环境中巨噬细胞功能及牙槽骨吸收的影响研究  

Effect and mechanism of Lactobacillus reuteri extracts on the macrophage function and alveolar bone resorption in periodontitis microenvironment

作　　者：刘奕彤 韩楠楠 杜鹃 郭力嘉[2] 刘怡[1] LIU Yi-tong;HAN Nan-nan;DU Juan;GUO Li-jia;LIU Yi(Department of Periodontics,Beijing Stomatological Hospital,Capital Medical University,Beijing 100050,China;Department of Orthodontics,Beijing Stomatological Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京口腔医院牙周科,北京100050 [2]首都医科大学附属北京口腔医院正畸科,北京100050

出　　处：《口腔颌面修复学杂志》2024年第1期6-14,共9页Chinese Journal of Prosthodontics

基　　金：国家重点研发计划(项目编号:2022YFC2504200);国家自然科学基金(项目编号:82201053);北京口腔医院青年科技创新人才培育计划(项目编号:YSP202105);北京市医管局青年人才培养计划“青苗”计划(项目编号:QML20231506)。

摘　　要：目的:探讨罗伊氏乳杆菌超声提取物对牙周炎微环境中巨噬细胞功能的影响。方法:体外筛选罗伊氏乳杆菌提取物干预小鼠巨噬细胞系RAW264.7的最佳浓度。随后将细胞分为PBS对照组、LPS组和LPS+罗伊氏乳杆菌提取物组,利用real-time PCR、ELISA和免疫荧光染色检测各组细胞的炎性巨噬细胞标志物的表达水平,利用Western blot对其机制进行探讨。体内建立大鼠丝线结扎牙周炎模型,观察罗伊氏乳杆菌提取物局部注射对于牙周骨组织破坏的治疗作用。结果:体外结果表明50μg/mL的罗伊氏乳杆菌提取物通过抑制NF-κB信号通路p-IκK和p-NF-κB蛋白的磷酸化(P<0.01),下调了LPS诱导的炎性巨噬细胞inos(P<0.05),Il1b(P<0.01)的基因表达水平,并抑制了LPS诱导的巨噬细胞TNF-α的细胞因子表达(P<0.05)。体内结果表明,罗伊氏乳杆菌提取物通过抑制牙周局部炎症进而下调破骨细胞的数量(P<0.01)和阳性面积(P<0.05),相较于牙周炎造成的骨吸收(P<0.01),罗伊氏乳杆菌提取物治疗减少了牙槽骨的吸收高度(P<0.05)。结论:罗伊氏乳杆菌提取物抑制炎症微环境中巨噬细胞的炎症因子表达,对牙周炎组织中牙槽骨的吸收破坏起到了保护作用。Objective:To investigate the effect and mechanism of the Lactobacillus reuteri extracts on the macrophages functions in the periodontitis microenvironment.Methods:The optimal concentration of Lactobacillus reuteri extract solution in vitro was explored using RAW264.7.Subsequently,the cells were divided into the PBS control group,the LPS group and the LPS+Lactobacillus reuteri extracts group.The expression levels of multiple inflammatory markers were detected by real-time PCR,ELISA and immunofluorescence staining,and the regulatory mechanism was explored by western blot.In addition,the regulatory effect of Lactobacillus reuteri extracts were investigated in vivo in the rat periodontitis model.Results:50μg/mL Lactobacillus reuteri extracts solution down-regulated the expression of LPS�induced inos(P<0.05)and Il1b(P<0.01)in RAW264.7 by inhibiting the phosphorylation of p-IκK and p-NF-κB proteins in NF-κB signaling pathway(P<0.01).The expression of TNF-αin RAW264.7 induced by LPS was also inhibited by Lactobacillus reuteri extracts(P<0.05).In vivo results showed that Lactobacillus reuteri extracts reduced the number of osteoclasts(P<0.01)and the TRAP positive area(P<0.05)in periodontitis tissues.Comparing with the bone resorption caused by periodontitis(P<0.01),Lactobacillus reuteri extracts reduced the resorption height of alveolar bone(P<0.05).Conclusion:Lactobacillus reuteri extracts inhibit the LPS-induced inflammatory macrophages activation via NF-κB signaling pathway,and play a protective role in the absorption and destruction of alveolar bone tissue under the inflammatory microenvironment.

关 键 词：罗伊氏乳杆菌 巨噬细胞 炎症 骨吸收 NF-ΚB信号通路 

分 类 号：R781.42[医药卫生—口腔医学]