通痹灵对CIA大鼠软骨细胞凋亡及其调控基因p53、Bcl-2表达作用的比较研究

目的探讨中药通痹灵在类风湿关节炎(RA)发病中对软骨细胞凋亡及其基因p53、Bcl-2调控的作用机制.方法采用大鼠CIA模型,分别予通痹灵、甲氨蝶呤片(MTX)、雷公藤治疗36 d,关节软骨免疫组化染色,计算机图像分析系统对软骨p53、Bcl-2基因蛋白及细胞凋亡的表达进行分析,比较各组差异.结果 CIA大鼠软骨细胞中存在高度活跃的p53蛋白表达,通痹灵可以下调细胞中过度活跃的p53蛋白表达,MTX高剂量及雷公藤高剂量的作用不显著;大鼠软骨细胞Bcl-2蛋白表达呈抑制状态,通痹灵低剂量组明显上调Bcl-2蛋白的低表达水平,其次为雷公藤高剂量组;大鼠软骨细胞存在明显的凋亡亢进,通痹灵明显下调细胞凋亡水平.结论通痹灵能通过调整过度表达的p53蛋白与低表达的Bcl-2蛋白,抑制软骨细胞的过度凋亡;MTX与雷公藤对凋亡的作用与Bcl-2基因家族的调控密切相关,与p53相关基因的相关性不显著.

Comparison of effects of Tongbiling on apoptosis of cartilage cells in CIA rats and expressions of p53, Bcl-2 genes

Objective To investigate the effective mechanism of Chinese drugs in cartilage cell apoptosis and the regulation of p53, Bcl-2 genes of rheumatoid arthritis (RA). Methods The CIA model rats were divided and treated with Tonbiling, Methotrenxate Tablets (MTX Tablets) and Tripterygium wilfordii Hook respectively for 36 days. The immunohistochemical stain was used in cartilage cells and computer imagine analysis system was used to analyze the expression of p53, Bcl-2 gene proteins and cell apoptosis. The differences in different groups were compared. Results There was an excessive protein expression of p53 in CIA rat cartilage cells. Tongbiling could inhibit the excessive expression of p53, while a high-dose of MTX Tablets or Tripterygium wilfordii Hook had no obvious effect. The protein expression of Bcl-2 in CIA rat cartilage cells was in an inhibitory state. With low-dose of Tongbiling the protein expression of Bcl-2 increased obviously and that in the high-dose of Tripterygium wilfordii Hook increased too. There was an accentuation in CIA rat cartilage cells. Tongbiling could low-regulate the level of cell apoptosis. Conclusion Tongbiling can inhibit the excessive apoptosis of cartilage cells through regulating the excessive protein expression of p53 and low protein expression of Bcl-2 . The effects of MTX Tablets and Tripterygium wilfordii Hook are related closely to the regulation of gene family, but not to p53 relative gene.