基于Notch1信号通路调控巨噬细胞极化改善佐剂关节炎大鼠关节炎症的机制  

The mechanism of regulating macrophage polarization based on Notch1 signaling pathway to improve joint inflammation in adjuvant arthritis rats

作　　者：程静 万磊[2] 赵磊 李舒 李方泽 胡赛赛 陈莹莹 CHENG Jing;WAN Lei;ZHAO Lei;LI Shu;LI Fang-ze;HU Sai-sai;CHEN Ying-ying(Anhui University of Traditional Chinese Medicine,Hefei 230031,China;Department of Rheumatology,the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学,安徽合肥230031 [2]安徽中医药大学第一附属医院风湿科,安徽合肥230031

出　　处：《海南医学院学报》2023年第23期1780-1785,共6页Journal of Hainan Medical University

基　　金：国家自然基金面上项目(81973655,82274501);国家中医药管理局第七批全国老中医药专家学术经验继承项目[国中医药人教函(2022)76号];安徽省高校优秀青年人才支持计划项目[皖教秘人(2022)11号];新安医学教育部重点实验室(2020xayx04);高校优秀青年人才支持计划项目[皖教秘人(2022)11号];安徽省高等学校质量工程项目省级教育教学改革研究项目(2022jyxm883)。

摘　　要：目的:探讨Notch1/Jagged1/RBP-Jκ/Hes1信号轴通过调控巨噬细胞向M2型极化对佐剂性关节炎(AA)大鼠炎症反应的影响。方法:大鼠随机分成3组(6只):健康组(NC)、模型组(MC)及Notch1抑制剂组(FLI),在致炎后12 d开始给予药物,并于30 d后检测各组大鼠的关节炎指数(AI)、右后足关节肿胀度(E);采用流式细胞仪测定CD80+和CD163+细胞在大鼠外周血巨噬细胞中的表达量;ELISA检测大鼠血清中IL-4、IL-10、IL-1β、及TNF-α的水平;Western Blot检测大鼠关节滑膜组织中Notch1、Jagged1、RBP-Jκ、Hes1蛋白的表达。结果:MC组AA大鼠的E及AI均明显高于NC组(P<0.01);CD80+细胞的表达显著高于对照组(P<0.01),而CD163+细胞的表达显著低于对照组(P<0.01);IL-1β、TNF-α表达水平明显升高(P<0.01), IL-4、IL-10表达水平明显降低(P<0.01);Notch1、RBP-Jκ、Jagged1、Hes1蛋白的表达量明显增加(P<0.01);与MC组比较,Notch1抑制剂组大鼠E、AI显著降低(P<0.01);CD80+细胞表达明显降低(P<0.01),CD163+细胞表达明显升高(P<0.01);IL-1β、TNF-α表达水平明显降低(P<0.05),IL-4、IL-10显著增加(P<0.01);Notch1、Jagged1、Hes1、RBP-Jκ蛋白表达量显著降低(P<0.05)。相关性分析表明,CD80+与Notch1、Jagged1、Hes1、RBP-Jκ呈正相关(P<0.01),CD163+与上述蛋白的表达呈负相关(P<0.01)。结论:阻断Notch1/Jagged1/RBP-Jκ/Hes1信号轴可以调控巨噬细胞向M2型极化,减轻AA大鼠的炎症反应。Objective:To study the impact of the Notch1/Jagged1/RBP⁃Jκ/Hes1 signaling pathway on macrophage polar⁃ization and its role in modulating the inflammatory response in rats with adjuvant arthritis(AA).Methods:The rats were random⁃ly divided into three groups(6 rats):the healthy group(NC),the model group(MC),and the Notch1 inhibitor group(FLI).Medication was administered after 12 days of inducing inflammation.After 30 days,the arthritis index and degree of swelling in the right hind foot joint were measured in each group.The expression levels of CD80+and CD163+cells in peripheral blood macro⁃phages of rats were analyzed by flow cytometry.The levels of IL⁃4,IL⁃10,IL⁃1β,and TNF⁃αin rat serum were measured by enzyme⁃linked immunosorbent assay.The expression of Notch1,Jagged1,RBP⁃Jκ,and Hes1 proteins in rat synovial tissue was detected using Western blot.Results:The degree of swelling and arthritis index in the MC group rats with AA were significantly higher than those in the NC group(P<0.01).CD80+cell expression was significantly higher compared to the control group(P<0.01),while CD163+cell expression was significantly lower than the control group(P<0.01).IL⁃1βand TNF⁃αexpression lev⁃els were significantly elevated(P<0.01),whereas IL⁃4 and IL⁃10 expression levels were significantly decreased(P<0.01).Notch1,RBP⁃Jκ,Jagged1,and Hes1 protein expression levels were significantly increased(P<0.01).In comparison to the MC group,the rats in the Notch1 inhibitor group exhibited a significant reduction in toe swelling and arthritis index(P<0.01).CD80+cell expression was significantly decreased(P<0.01),while CD163+cell expression was significantly increased(P<0.01).IL⁃1βand TNF⁃αexpression levels were significantly decreased(P<0.05),whereas IL⁃4 and IL⁃10 levels were significantly increased(P<0.01).Notch1,Jagged1,Hes1,and RBP⁃Jκprotein expression levels were significantly decreased(P<0.05).Correlation analysis revealed a positive association between CD80+and Notch1,Jagged1,Hes1,

关 键 词：佐剂关节炎 Notch1/Jagged1/RBP-Jκ/Hes1信号轴 巨噬细胞极化 

分 类 号：R593.22[医药卫生—内科学]