摘要
目的探讨局部应用傅山风湿外治方对胶原诱导性关节炎(Collagen-induced arthritis,CIA)大鼠炎性细胞因子和Notch2表达的影响。方法雌性Wistar大鼠36只,随机分为正常组10只,造模组26只,成功构建CIA模型20只,随机平均分配为模型组、傅山风湿外治方组各10只。第14天开始,傅山风湿外治方组将0.4 mL药膏均匀涂抹于大鼠踝关节。正常组和模型组给予等体积生理盐水局部涂抹。每天更新2次,连续42天。每周观察大鼠关节肿胀情况进行关节炎指数评分。干预结束后,取大鼠腹主动脉血、踝关节、脾脏和肝肾组织。HE染色评估关节炎严重程度及踝关节和肝肾组织病理变化;ELISA检测血清炎性细胞因子表达;qRT-PCR和Western blot检测踝关节滑膜和脾脏Notch2、Delta-like配体蛋白1(Delta-like ligand protein-1,DLL1)和核因子-κBp65(Nuclear factor-κBp65,NF-κBp65)mRNA和蛋白表达,并利用免疫组织化学法检测其在踝关节的阳性表达;检测各组大鼠血清肝肾功能水平变化。结果与正常组相比,模型组关节炎指数评分升高(P<0.01),关节损伤增加,病理学评分升高(P<0.01),血清炎性细胞因子TNF-α、IL-6、IL-17和IFN-γ水平升高(P<0.01),大鼠踝关节和脾脏Notch2、DLL1和NF-κBp65 mRNA和蛋白的表达升高(P<0.01),踝关节阳性表达增多(P<0.01);与模型组相比,傅山风湿外治方组关节炎指数评分下降(P<0.05,P<0.01),关节损伤减轻,病理学评分降低(P<0.01),血清炎性细胞因子TNF-α、IL-6、IL-17和IFN-γ水平下降(P<0.01),大鼠踝关节和脾脏中Notch2、DLL1和NF-κBp65 mRNA和蛋白的表达降低(P<0.01),踝关节阳性表达减少(P<0.01),肝肾组织均未见明显的病理损伤,血清天冬氨酸转氨酶(Aspartate transaminase,AST)、丙氨酸转氨酶(Alanine transaminase,ALT)、血尿素氮(Blood urea nitrogen,BUN)、肌酐(Creatinine,Cr)水平无显著变化(P>0.05)。结论傅山风湿外治方可缓解CIA大鼠的关节炎症状,减轻关节损�
Objective To explore the effect of topical application of Fushan rheumatism external prescription on inflammatory cytokines and notch2 pathway in rats with collagen-induced arthritis(CIA).Methods 36 female Wistar rats were randomly divided into normal group(n=10)and model group(n=26).CIA model was successfully established in 20 rats,which were randomly divided into model group(n=10)and Fushan rheumatism external prescription group(n=10).Treatment was initiated on day 14,and 0.4 mL ointment was evenly applied to the ankle joints of rats in Fushan rheumatism external prescription group.Normal group and model group were topical smeared with the same volume of normal saline.Activity was taken twice a day for 42 days.The joint swelling of rats was observed every week and the arthritis index was scored.Thereafter blood was collected from abdominal aorta,and then ankle joint,spleen,liver,and kidney of rats were taken out after the end of the interventions.The severity of arthritis and the pathological changes of ankle joint,liver and kidney were evaluated by HE staining;Inflammatory cytokines expression in serum were detected by ELISA;The expression of Notch2,Delta-like ligand protein 1(delta-like ligand protein-1,DLL1)and nuclear factor-κ Bp65(nuclear factor-κ Bp65,NF-κ Bp65)mRNA and protein in synovium of ankle joints and spleen were detected by qRT-PCR and Western blot,and its positive expression in ankle joints were detected by immunohistochemical method;The changes of liver and kidney function of rats in each group were detected in serum.Results Compared with normal group,the arthritis index score in model group were increased(P<0.01),joint injury and pathological score were increased(P<0.01),the levels of inflammatory cytokines TNF-α,IL-6,IL-17 and IFN-γ in serum were increased(P<0.01),the expression of Notch2,DLL1 and NF-κBp65 mRNA and protein in joints and spleen were increased(P<0.01),and the positive expression in joints were also increased(P<0.01);Compared with model group,the arthritis index score in F
作者
高玉亭
李振
赵彩虹
赵雨薇
王泽
郝慧琴
Gao Yuting;Li Zhen;Zhao Caihong;Zhao Yuwei;Wang Ze;Hao Huiqin(College of Basic Medical Sciences,Shanxi University of Chinese Medicine,Jinzhong 030619,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第7期2604-2611,共8页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
山西省科学技术厅山西省重点研发计划项目社会发展方面(201803D31084):傅山风湿外治方治疗风寒湿型类风湿关节炎有效性及安全性研究,负责人:郝慧琴
山西省科学技术厅山西省基础研究计划青年科技研究基金(201901D211534):基于Noctch1/Jagged通路探讨傅山风湿外治方防治类风湿关节炎的作用机制,负责人:高玉亭
山西中医药大学博士科研启动基金(2022BK02):山西医家方剂挖掘及傅山风湿外治方对类风湿关节炎的治疗作用和机制研究,负责人:赵雨薇。
