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论文摘要

王枣子总黄酮对佐剂性关节炎大鼠的治疗作用及机制研究

Effect of Flavonoids from Wang Zaizi on Adjuvant Arthritis in Rats and Its Mechanism

作者:缪成贵, 熊友谊, 秦梅颂等

Author:MIAO Cheng-gui, XIONG You-yi, QING Mei-song. et al

收稿日期:          年卷(期)页码:2018,49(3):374-379

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:王枣子总黄酮 佐剂性关节炎 miR-152 成纤维样滑膜细胞 IL-1

Key words:Total flavonoids of Isodon amethystoides (Ben-th) Cy Wu et Hsuan Adjuvant arthritis miR-152 Fibroblast like synoviocytes IL-1

基金项目:

中文摘要

目的 研究王枣子中总黄酮(TFIA)对佐剂性关节炎(AA)的治疗作用及机制。方法 采用注射完全弗氏佐剂制备AA模型大鼠后随机分为4组:AA组、AA+TFIA 50 mg/kg组、AA+TFIA 100 mg/kg组、AA+TFIA 150 mg/kg组,每组10只。另设空白对照组大鼠,不造模(n=10)。造模后4 d,TFIA各剂量组大鼠采用TFIA灌胃治疗,分别按上述3个剂量灌胃,每天1次,共24 d,空白对照组和AA组灌胃生理盐水。治疗第12天、16天、20天、24天采用大鼠关节炎评分法评价TFIA对AA大鼠的治疗作用。治疗第24天处死各组大鼠,分离大鼠滑膜组织,原代培养各组大鼠成纤维样滑膜细胞(FLS),ELISA检测各组FLS中白细胞介素(IL-1)表达,MTT检测各组FLS增殖活力,real time qPCR检测各组FLS miR-152和经典Wnt信号通路关键基因〔β-catenin、细胞周期蛋白基因(ccnd1)〕表达。向AA大鼠FLS转染miR-152 mimics和对照NC mimics,向TFIA 100 mg/kg剂量组大鼠FLS转染miR-152 inhibitors和对照NC inhibitors,36 h后检测miR-152、β-catenin、ccnd1、IL-1表达和FLS的增殖。结果 TFIA各剂量灌胃治疗均能抑制AA大鼠关节炎评分,抑制β-catenin、ccnd1、IL-1表达和FLS增殖活力(PP>0.05),与空白对照组相比,各剂量组均未恢复至正常水平(PPccnd1、IL-1表达,抑制FLS增殖(Pccnd1、IL-1表达,促进了FLS增殖(P

英文摘要

Objective The therapeutic effect and mechanism of total flavonoids in Isodon amethystoides (Ben-th) Cy Wu et Hsuan (TFIA) on adjuvant arthritis (AA) were investigated. Methods AA model rats were set and complete Freund’s adjuvant injection, randomly divided into 4 groups: AA group, AA+TFIA 50 mg/kg group, AA+TFIA 100 mg/kg group, AA+TFIA 150 mg/kg group, and each group has 10 rats. Blank control group was set without modeling (n=10). Four days post-modeling rats in each TFIA groups were treated once a day with TFIA at 50 mg/kg, 100 mg/kg and 150 mg/kg for 24 d, and rats in blank control and AA groups were given saline as control. At the 12th day, 16th day, 20th day and 24th day of treatment, the effect of TFIA on AA rats was evaluated by rat arthritis score. Then the rats were sacrificed on the 24th day of treatment, and the synovial tissue of rats was isolated and the fibroblast-like synoviocytes (FLS) were primary cultured. The expressions of IL-1 in FLS was detected by ELISA, the FLS proliferation activity was detected by MTT assay, and the expression of miR-152, β-catenin and cyclin D1 gene (ccnd1) were detected by real time qPCR. MiR-152 mimics and NC mimics (control) were transfected into FLS in AA rats, and miR-152 inhibitors and NC inhibitors (control) were transfected into FLS in AA+TFIA 100 mg/kg group rats. The expressions of miR-152, β-catenin,ccnd1,IL-1 and FLS proliferation were detected 36 h post-transfection. Results TFIA significantly inhibited the arthritis socre of rats and the expressions of β-catenin,ccnd1,IL-1 and the proliferation of FLS in AA rats (PPPccnd1,IL-1 and the proliferation of FLS (Pccnd1,IL-1 and the proliferation of FLS. Conclusion TFIA has a certain therapeutic effect on AA rats via the up-regulation of miR-152 expression, possibly affecting the classical Wnt signaling pathway.

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