摘要
目的基于Notch1/Jagged1/RBP-Jκ/Hes1信号通路调控巨噬细胞极化探讨复方雷公藤制剂对佐剂关节炎大鼠关节炎症的影响。方法将30只雄性SD大鼠随机分为正常组、模型组、复方雷公藤组、甲氨蝶呤组、Notch1抑制剂组,每组6只,除正常组外,其余组大鼠构建佐剂关节炎模型。致炎后第19天开始,复方雷公藤组给予340 mg/L复方雷公藤制剂混悬液灌胃,每天1次;甲氨蝶呤组给予0.95 mg/L甲氨蝶呤混悬液灌胃,每周1次;Notch1抑制剂组给予0.56 mg/L Notch1抑制剂混悬液尾静脉注射,隔天1次;正常组及模型组给予生理盐水灌胃,每天1次。各组均连续干预30 d。检测各组大鼠足趾肿胀度、关节炎指数;末次灌胃结束后,HE染色观察关节滑膜组织病理学形态,ELISA法检测血清IL-1β、IL-4、IL-10、IL-13、TNF-α水平,流式细胞术检测外周血炎症极化标志物CD86、CD206表达情况,免疫组化及免疫印迹法检测关节滑膜组织中Notch1、Jagged1、RBP-Jκ、Hes1蛋白表达情况。结果与正常组比较,模型组大鼠足趾肿胀度、关节炎指数、外周血CD86表达占比及血清IL-1β、TNF-α水平和关节滑膜组织中Notch1、Jagged1、RBP-Jκ、Hes1蛋白相对表达占比均明显升高(P均<0.05),外周血CD206表达占比及血清IL-4、IL-10、IL-13水平均明显降低(P均<0.05);与模型组比较,复方雷公藤组、甲氨蝶呤组、Notch1抑制剂组大鼠足趾肿胀度、关节炎指数(除Notch1抑制剂组)、外周血CD86表达占比及血清IL-1β、TNF-α水平和关节滑膜组织中Notch1、Jagged1、RBP-Jκ、Hes1蛋白相对表达占比均明显降低(P均<0.05),外周血CD206表达占比及血清IL-4、IL-10、IL-13水平均明显升高(P均<0.05)。结论复方雷公藤制剂可有效减轻佐剂关节炎大鼠关节炎症反应,其机制可能与抑制Notch1/Jagged1/RBP-Jκ/Hes1信号通路轴调控巨噬细胞极化,抑制促炎细胞因子的分泌有关。
Objective It is to observe the effects of compound Tripterygium wilfordii preparation on joint inflammation in adjuvant arthritis rats via regulating macrophage polarization through regulating Notch1/Jagged1/RBP-Jκ/Hes1 signaling pathway.Methods Thirty male SD rats were randomly divided into normal group,model group,compound Tripterywilfornia preparation group,methotrexate group,Notch1 inhibitor group,with 6 rats in each group.The rats of all groups except for the normal group were prepared to establish models of adjuvant arthritis(AA).The dosage was administered on the 19th day after inflammation,and the degree of toe swelling and arthritis index of each group were detected 30 days after administration.Beginning on the 19th day after the onset of inflammation,the compound Tripterywilfornia preparation group was given 340 mg/L of compound Tripterywilfornia preparation suspension by gavage,once per day;the methotrexate group was given 0.95 mg/L of methotrexate suspension by gavage,once per week;the Notch1 inhibitor group was given 0.56 mg/L of Notch1 inhibitor suspension by caudal vein injection,every other day;the normal group and model group were given normal saline by gavage,once per day.All the groups were continuously treated for 30 days.The toe swelling and arthritis index of rats in each group were detected;at the end of the last gavage,the histopathological morphology of synovial joint tissues was observed by HE staining,the serum levels of IL-1β,IL-4,IL-10,IL-13,and TNF-α were detected by ELISA,the expressions of inflammation polarization markers CD86 and CD206 in peripheral blood were detected by flow cytometry,and the expressions of Notch1,Jagged1,RBP-Jκ,and Hes1 proteins in synovial joint tissues were detected by immunohistochemistry and immunoblotting.Results Compared with the normal group,the toe swelling,arthritis index,CD86 percentage in peripheral blood,levels of IL-1βand TNF-αin serum,and relative expressions of Notch1,Jagged1,RBP-Jκ,and Hes1 proteins in synovial tissues of rats in the model group were all significantly increased(all P<0.05),while CD206 percentage in peripheral blood and the levels of IL-4,IL-10 and IL-13 in serum were significantly decreased(all P<0.05).Compared with the model group,the toe swelling,arthritis index(except for the Notch 1 inhibitor group),CD86 percentage in peripheral blood,levels of IL-1βand TNF-α in serum,and relative expressions of Notch1,Jagged1,RBP-Jκ,and Hes1 proteins in synovial tissues of rats in the compound Tripterywilfornia preparation group,methotrexate group,Notch1 inhibitor group were all significantly decreased(all P<0.05),while CD206 percentage in peripheral blood and the levels of IL-4,IL-10 and IL-13 in serum were significantly increased(all P<0.05).Conclusion Compound Tripterygium wilfordii preparation can effectively alleviate joint inflammation in AA rats,and the mechanism may be related to regulate macrophage polarization and inhibit the secretion of pro-inflammatory cytokines via inhibiting Notch1/Jagged1/RBP-Jκ/Hes1 signaling pathway.
作者
赵磊
万磊
刘健
黄传兵
贾光辉
朱子衡
胡恩钦
娄渊和
ZHAO Lei;WAN lei;LIU Jian;HUANG Chuanbing;JIA Guanghui;ZHU Ziheng;HU Enqin;LOU Yuanhe(JieShou People’s Hospital,Jieshou 236500,Anhui,China;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,Anhui,China;Key Laboratory of Education Ministry of Xin’an Medicine,Hefei 230038,Anhui,China)
出处
《现代中西医结合杂志》
CAS
2024年第5期602-608,共7页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
国家自然科学基金面上项目(81973655,82274501)
安徽省阜阳市卫生健康委科研立项项目(FY2023-089)
新安医学与中医药现代化研究所揭榜挂帅项目(2023CXMMTCM020)
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)
安徽省自然科学基金项目(2308085MH291)
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)
安徽省高校自然科学研究项目(2023AH050814)
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)。
