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目的研究野菊花总黄酮(TFC)对佐剂性关节炎(AA)大鼠滑膜细胞凋亡的影响,并探讨其作用机理。方法20只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,电镜观察细胞形态改变,Western blotting分析大鼠滑膜细胞caspase-3活化片段蛋白的表达变化,annexin V荧光染色检测caspase-3特异性阻断剂对大鼠滑膜细胞凋亡的抑制作用。结果TFC能诱导大鼠滑膜细胞凋亡,电镜观察到典型的凋亡细胞核。大鼠滑膜细胞caspase-3活化片段蛋白表达随TFC浓度增加而明显升高,caspase-3特异性阻断剂可以明显减轻AA大鼠滑膜细胞凋亡。结论TFC可促进大鼠滑膜细胞凋亡而达到治疗AA作用,其机制与TFC诱导滑膜细胞凋亡及提高caspase-3的活化有关。
Objective To study the effect of total flavonoids (TFC) of chrysanthemum on the apoptosis of synovial cells in adjuvant arthritis (AA) rats and to explore its mechanism of action. Methods Twenty SD rats were injected with 0.1 ml Freund’s complete adjuvant into the right hind paw to replicate the AA model. After 24 days, the rat synoviocytes were isolated and cultured by tissue culture method. The morphological changes of the cells were observed by electron microscopy and analyzed by Western blotting. Expression changes of caspase-3 activation fragment protein in synoviocytes of rat were detected by annexin V fluorescence staining. The inhibitory effect of caspase-3 specific blocker on apoptosis of synoviocytes in rat was detected. Results TFC could induce apoptosis of synoviocytes in rats. Typical apoptotic nuclei were observed under electron microscope. The expression of caspase-3 activation fragment in synoviocytes of rats increased with the increase of TFC concentration. The caspase-3 specific blocker can significantly reduce the apoptosis of synoviocytes in AA rats. Conclusion TFC can promote apoptosis of synoviocytes in rats and achieve therapeutic effect of AA. Its mechanism is related to TFC-induced apoptosis of synovial cells and increase of caspase-3 activation.