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目的研究野菊花有效成分总黄酮(TFC)对佐剂性关节炎(AA)大鼠滑膜细胞凋亡的影响,探讨其治疗作用的机理。方法SD大鼠右后足跖皮内注射0.1ml弗氏完全佐剂复制AA模型;24d后组织块培养法分离培养大鼠膝关节滑膜细胞,用MTT法检测TFC对滑膜细胞增殖的影响;Hoechst 33258荧光染色,观察滑膜细胞核是否出现凋亡小体;琼脂糖凝胶电泳观察DNA碎片。结果MTT法显示TFC抑制滑膜细胞增殖,并且呈剂量依赖性趋势,药物作用24h的IC50为112mg/L;TFC能诱导滑膜细胞凋亡,电镜观察到典型的凋亡细胞核,电泳呈现出阶梯状条带,Hoechst 33258染色观察到细胞核出现凋亡小体。结论TFC可能会通过抑制AA大鼠滑膜细胞的增生,促进滑膜细胞的凋亡而达到治疗类风湿性关节炎的作用。
Objective To study the effect of total flavonoids (TFC), an effective component of chrysanthemum on the apoptosis of synoviocytes in adjuvant arthritis (AA) rats, and to explore the mechanism of its therapeutic effect. Methods The AA model was replicated by intradermal injection of 0.1 ml Freund’s complete adjuvant to the right hind paw of SD rats. After 24 days, the rat synoviocytes were isolated and cultured by tissue culture method. The effect of TFC on proliferation of synovial cells was detected by MTT assay. Hoechst 33258 fluorescence staining was used to observe whether apoptotic bodies appeared in synovial nucleus; DNA fragments were observed by agarose gel electrophoresis. Results MTT assay showed that TFC inhibited the proliferation of synovial cells in a dose-dependent manner. The IC50 of the drug at 24 h was 112 mg/L. TFC could induce apoptosis of synovial cells. Typical apoptotic nuclei were observed by electron microscopy and electrophoresis showed a ladder. Apoptotic bodies were observed in the nucleus by Hoechst 33258 staining. Conclusion TFC may be used to treat rheumatoid arthritis by inhibiting the proliferation of synovial cells in AA rats and promoting the apoptosis of synovial cells.