基于CX3CL1/NF-κB信号通路探讨补肾强督方抑制M1巨噬细胞极化治疗强直性脊柱炎的机制研究  

Mechanism of Bushen Qiangdu formula inhibiting M1 macrophage polarization by CX3CL1/NF⁃κB signaling pathway in the treatment of ankylosing spondylitis

作　　者：吴琳 李松倍[1] 苏晓庆 韩天然 李泽光[2] WU Lin;LI Songbei;SU Xiaoqing;HAN Tianran;LI Zeguang(Heilongjiang University of Chinese Medicine,Harbin 150040,China;First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040

出　　处：《海南医学院学报》2024年第19期1478-1485,共8页Journal of Hainan Medical University

基　　金：白求恩医学科学研究基金(TY144EN)。

摘　　要：目的:观察补肾强督方对CX3CL1/NF-κB信号通路介导的M1型巨噬细胞极化的影响,从而阐明其治疗强直性脊柱炎的分子机制。方法:采用脂多糖(LPS)诱导THP-1细胞构建细胞模型,并给予补肾强督方含药血清和CX3CL1抑制剂AZD8797进行干预。采用酶联免疫吸附试验检测细胞炎症因子(TNF-α、IL-6、IL-1β和IL-17)的含量;采用流式细胞术检测CD86的表达;采用蛋白印迹检测iNOS、破骨分化标记蛋白(TRAP、Calcitonin和p-NFATC1β)及CX3CL1/NF-κB信号通路中相关蛋白的表达;采用实时荧光定量逆转录聚合酶链反应检测iNOS mRNA的表达。结果:与模型组相比,与空白组相比,模型组巨噬细胞TNF-α、IL-6、IL-1β、IL-17的含量和CD86+巨噬细胞的比例显著增加,iNOS、CX3CL1、p-P65、p-IKKα/β、p-IkBα、TRAP、Calcitonin和p-NFATC1β蛋白和iNOS mRNA的表达显著上调,差异均具有统计学意义(P<0.01)。与模型组相比,补肾强督方能够显著降低巨噬细胞TNF-α、IL-6、IL-1β、IL-17的含量和CD86+巨噬细胞的比例,下调iNOS、CX3CL1、p-P65、p-IKKα/β、p-IkBα、TRAP、Calcitonin和p-NFATC1β蛋白和iNOS mRNA的表达,差异均具有统计学意义(P<0.05)。结论:补肾强督方通过抑制CX3CL1/NF-κB信号通路的激活,从而抑制巨噬细胞M1型极化,实现抑制炎症和破骨细胞的分化的作用。Objective:To observe the effect of Bushen Qiangdu formula on CX3CL1/NF⁃κB signaling pathway mediated po⁃larization of M1 macrophages and elucidate its molecular mechanism in treating ankylosing spondylitis.Methods:Lipopolysaccha⁃ride(LPS)⁃induced THP⁃1 cells were used to construct a cell model,and Bushen Qiangdu medicated serum and CX3CL1 inhibi⁃tor AZD8797 were given for intervention.The levels of inflammatory factors(TNF⁃α,IL⁃6,IL⁃1βand IL⁃17)were detected by enzyme⁃linked immunosorbent assay.CD86 expression was detected by flow cytometry.Expression of iNOS,osteoblast differenti⁃ation marker proteins(TRAP,calcitonin and p⁃NFATC1β)and the CX3CL1/NF⁃κB pathway was assessed by Western blot.The expression of iNOS mRNA was quantified by real⁃time quantitative polymerase chain reaction(qRT⁃PCR).Results:Com⁃pared with the control group,the content of TNF⁃α,IL⁃6,IL⁃1β,IL⁃17 in macrophages and the proportion of CD86+macro⁃phages in the model group were significantly increased,and iNOS,CX3CL1,p⁃P65,p⁃IkKα/β,p⁃IkBα,TRAP,calcitonin and p⁃NFATC1βprotein and iNOS mRNA expression were significantly upregulated,all of which were statistically significant(P<0.01).Bushen Qiangdu-medicated serum significantly decreased the content of TNF⁃α,IL⁃6,IL⁃1β,IL⁃17 and the proportion of CD86+macrophages,and down⁃regulated the expression of iNOS,CX3CL1,p⁃P65,p⁃IKKα/β,p⁃IkBα,TRAP,Calcitonin and p⁃NFATC1βproteins and iNOS mRNA expression,which showed statistically significant differences compared with the model group(P<0.05).Conclusion:Bushen Qiangdu formula inhibits M1 polarization of macrophages by inhibiting CX3CL1/NF⁃κB signaling pathway,then playing a role of inhibiting inflammation and osteoclast differentiation in the treatment of ankylosing spon⁃dylitis.

关 键 词：补肾强督方 CX3CL1/NF-κB信号通路 巨噬细胞极化 破骨细胞 分子机制 

分 类 号：R285[医药卫生—中药学]