基于miR-486-5p、巨噬细胞极化探讨新风胶囊改善佐剂性关节炎大鼠关节炎症的作用机制  

Exploring the mechanism of action of Xinfeng capsule to improve joint inflammation in rats with adjuvant arthritis based on miR⁃486⁃5p and macrophage polarization

作　　者：李柯霏 万磊[2,3,4] 刘磊 王丽文[1] 闫佩文 张玉如 LI Kefei;WAN Lei;LIU Lei;WANG Liwen;YAN Peiwen;ZHANG Yuru(The First Clinical College of 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;Center for Xin'an Medicine and Modernization of Traditional Chinese Medicine of IHM,Hefei 230012,China;Key Laboratory of Xin'an Medical Education Ministry,Hefei 230012,China)

机构地区：[1]安徽中医药大学第一临床医学院,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 [3]新安医学与中医药现代化研究所,安徽合肥230012 [4]新安医学教育部重点实验室,安徽合肥230012

出　　处：《海南医学院学报》2024年第21期1617-1624,共8页Journal of Hainan Medical University

基　　金：国家自然科学基金面上项目(82274501);新安医学与中医药现代化研究所揭榜挂帅项目(2023CXMMTCM020,2023CXM-MTCM004);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];新安医学教育部重点实验室项目(2020xayx04);安徽省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:观察新风胶囊对佐剂性关节炎(adjuvant arthritis,AA)大鼠巨噬细胞极化及miR-486-5p的影响,探究新风胶囊改善AA大鼠炎症的可能机制。方法:32只SD大鼠被随机分为两个不同组别:正常组(NC组,8只)和造模组(24只),通过对造模组大鼠右后足跖皮内注射弗氏完全佐剂,成功构建AA大鼠模型后,进一步随机细分为3组:模型组(MC组,8只)、新风胶囊组(XFC组,8只)以及miR-486-5p抑制剂组(miR-486-5p inhibitor组,8只)。于炎症诱导后的第14天开始给予相应药物和试剂,NC组及MC组持续21天灌胃等量的生理盐水,然后检测各组AA大鼠足跖肿胀度(E)、关节炎指数(AI);采用酶联免疫吸附法测定(ELISA)检测大鼠细胞因子白细胞介素-23(interleukin-23,IL-23)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、IL-4及IL-13;流式细胞术检测大鼠外周血CD86、CD206表达;RT-qPCR检测大鼠关节滑膜miR-486-5p表达。结果:(1)与NC组比较,MC组大鼠E、AI及IL-23、TNF-α、CD86和miR-486-5p表达升高(P<0.01);IL-4、IL-13、CD206表达降低(P<0.01)。(2)与MC组比较,XFC组和miR-486-5p inhibitor组E、AI、IL-23、TNF-α、CD86和miR-486-5p表达降低(P<0.01);IL-4、IL-13、CD206表达升高(P<0.01)。(3)与miR-486-5p inhibitor组比较,XFC组IL-4、IL-13、CD206表达升高(P<0.01);IL-23、TNF-α、CD86和miR-486-5p表达降低(P<0.01);E、AI差异不明显(P>0.05)。结论:AA大鼠炎症与巨噬细胞极化和miR-486-5p表达有关,新风胶囊能明显改善AA大鼠关节炎症,其机制可能与抑制miR-486-5p表达,调节巨噬细胞极化有关。Objective:To observe the effects of Xinfeng capsule on macrophage polarization and miR-486-5p in adjuvant arthritis(AA)rats,and to explore the possible mechanism of Xinfeng capsule to improve inflammation in AA rats.Methods:Thirtytwo SD rats were randomly divided into two different groups:the normal group(NC group,8 rats)and the modeling group(24 rats).After the AA rat model was successfully constructed by intradermal injection of Fuchs′complete adjuvant into the right hindfoot metatarsal of the rats in the modeling group,the rats were further randomly subdivided into three groups:the modeling group(MC group,8 rats),the Xinfeng capsule group(XFC group,8 rats),and the miR-486-5p inhibitor group(miR-486-5p inhibitor group,8 pcs).The corresponding drugs and reagents were given from the 14th day after the induction of inflammation,and the NC group and MC group continued to gavage an equal amount of saline for 21 days,and then the foot-plantar swelling(E)and arthritis index(AI)of the AA rats in each group were detected;and the cytokines interleukin-23(IL-23),tumor necrosis factor-α(TNF-α),IL-4 and IL-13 and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA)in the rats;flow cytometry was used to detect the expression of CD86 and CD206 in peripheral blood of rats;RT-qPCR was used to detect the expression of miR-486-5p in synovial membrane of rats.Results:(1)Compared with the NC group,rats in the MC group showed elevated expression of E,AI and IL-23,TNF-α,CD86 and miR-486-5p(P<0.01);and decreased expression of IL-4,IL-13 and CD206(P<0.01).(2)Compared with the MC group,the expression of E,AI,IL-23,TNF-α,CD86 and miR-486-5p was decreased in the XFC group and miR-486-5p inhibitor group(P<0.01);the expression of IL-4,IL-13 and CD206 was elevated(P<0.01).(3)Compared with miR-486-5p inhibitor group,IL-4,IL-13,CD206 expression was elevated in XFC group(P<0.01);IL-23,TNF-α,CD86,and miR-486-5p expression was reduced(P<0.01);the difference of E and AI was not significant(P>0.05).Conclu

关 键 词：类风湿关节炎 新风胶囊 miR-486-5p 巨噬细胞极化 佐剂性关节炎大鼠 

分 类 号：R285.5[医药卫生—中药学]