去氢吴茱萸碱在治疗类风湿关节炎中的作用和机制研究  

Study on the role and mechanism of dehydroevodiamine in the treatment of rheumatoid arthritis

作　　者：张婷婷 李希凤 冯晴雯 王林 宋冠华 ZHANG Tingting;LI Xifeng;FENG Qingwen;WANG Lin;SONG Guanhua(Shandong First Medical University(Shandong Academy of Medical Sciences),Jinan 250000,China)

机构地区：[1]山东第一医科大学(山东省医学科学院),山东济南250000

出　　处：《临床医学研究与实践》2023年第16期1-4,18,共5页Clinical Research and Practice

基　　金：山东第一医科大学省级大学生创新创业训练计划项目(No.S202110439153)。

摘　　要：目的研究去氢吴茱萸碱(DHED)对类风湿关节炎(RA)的影响及其作用机制。方法将培养的滑膜成纤维样细胞(FLS)分为对照组和DHED组。体外实验采用CCK-8检测细胞增殖;Annexin V-FITC/PI双染检测细胞凋亡;Transwell和划痕实验检测细胞侵袭和迁移;酶联免疫吸附测定(ELISA)检测炎性因子分泌水平;双荧光素酶报告基因法检测抗核因子-κB(NF-κB)信号通路活性;Western blot实验检测P65的磷酸化。体内实验以80只6~8周龄DAB/1小鼠[体重(20.24±2.51)g]为研究对象,以胶原诱导性关节炎(CIA)小鼠为模型,将其分为对照组、阳性药物处理组[依那西普组(400μg/kg)、DHED低剂量组(10 mg/kg)和DHED高剂量组(20 mg/kg)],验证DHED对小鼠关节炎进展的影响。结果DHED可显著降低RA FLS增殖活性,促进凋亡,抑制侵袭与迁移,下调肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)介导的促炎因子分泌,抑制P65磷酸化水平,显著缓解关节炎小鼠疾病进展。结论DHED可显著改善RA FLS异常表型,缓解RA疾病进展,其潜在作用机制可能与抑制NF-κB信号通路有关。Objective To study the effect of dehydroevodiamine(DHED)on rheumatoid arthritis(RA)and its mechanism.Methods The cultured fibroblast like synoviocytes(FLS)were divided into control group and DHED group.CCK-8 was used to detect cell proliferation in vitro;apoptosis was detected by Annexin V-FITC/PI double staining;Transwell and scratch experiments were used to detect cell invasion and migration;the secretion level of inflammatory factors were detected by enzyme linked immunosorbent assay(ELISA);the activity of anti-nuclear factor-kappa B(NF-κB)signaling pathway was detected by dual luciferase reporter gene assay;the phosphorylation of P65 was detected by Western blot experiment.In vivo experiments,80 DAB/1 mice aged 6-8 weeks[body weight(20.24±2.51)g]were selected as the research objects,and collagen induced arthriti(CIA)mice was used as the model.They were divided into control group and positive drug treatment group[etanercept group(400μg/kg),DHED low dose group(10 mg/kg)and DHED high dose group(20 mg/kg)]to verify the effect of DHED on the progression of arthritis in mice.Results DHED can significantly reduce the proliferation activity of RA FLS,promote apoptosis,inhibit invasion and migration,down-regulate the secretion of pro-inflammatory factors mediated by tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β),inhibit the phosphorylation level of P65,and significantly relieve the progression of arthritis in mice.Conclusion DHED can significantly improve the abnormal phenotype of RA FLS and alleviate the progression of RA,and its potential mechanism may be related to the inhibition of NF-κB signaling pathway.

关 键 词：去氢吴茱萸碱 类风湿关节炎 炎性因子 滑膜成纤维样细胞 核因子-ΚB信号通路 

分 类 号：R593.22[医药卫生—内科学]