双藤汤醇提取物对寒痹型胶原诱导型关节炎大鼠凋亡调节作用

Effect of Alcohol Extract of Shuangteng Decoction on Apoptosis of Collagen-induced Arthritis Rats Aggravated by Cold

目的：观察双藤汤对寒痹型胶原诱导型关节炎大鼠抗凋亡作用。方法：随机选取雄性SD大鼠分为空白组、模型组、双藤汤60、40、20 g/kg组、青藤碱28 mg/kg组,每组10只。除空白组外,其余各组构建模型。连续给药4周后测量动物足跖厚度;HE染色分析脾脏病理变化;免疫组化法检测脾脏Bax和Bcl-2蛋白表达,Western-blotting检测脾脏Caspase-1,Caspase-3和CytC蛋白表达。结果：与模型组比较,空白组、双藤汤60 g/kg和40 g/kg组和青藤碱28 mg/kg组足跖部肿胀程度下降;脾脏病理切片显示空白组、双藤汤60 g/kg和40 g/kg组和青藤碱28 mg/kg抑制脾小体萎缩减少并促进其发育;免疫组织法显示双藤汤60、40、20 g/kg组、青藤碱28 mg/kg组Bax蛋白表达下降,Bcl-2蛋白表达上调;蛋白印迹显示双藤汤60、40、20 g/kg组、青藤碱28 mg/kg组Caspase-1、Caspase-3、CytC蛋白表达受到不同程度抑制。结论：双藤汤可能通过调节凋亡途径中Bax、Bcl-2、Caspase-1、Caspase-3、CytC蛋白表达起到对寒痹型胶原诱导关节炎大鼠炎症的抑制作用。

Objective：To observe the effect of Shuangteng Decoction on anti-apoptosis of collagen-induced arthritis rats aggravated by cold. Method： Male SD rats were randomly selected and divided into sham group, model group, 60 g/kg,40 g/kg and 20 g/kg groups of Shuangteng Decoction and 28 mg/kg group of sinomenine,10 in each group. Collagen-induced arthritis was constructed in the other groups except the sham group. After 4 weeks of continuous administration,the thicknesses of vola pedis were measured and the pathological changes were observed by spleen HE staining. Bax and Bcl-2 protein expressions in spleen were detected by immunohistochemical method. Western-blotting was used to detect the expressions of Caspase-1,Caspase-3,CytC protein in spleen. Result：Compared with the model group,the swelling degree of vola pedis were inhibited in sham group,Shuangteng Decoction 60 g/kg group and 40 g/kg group and sinomenine 28 mg/kg group. The spleen pathology showed spleen corpuscle atrophy was inhibited in sham group, Shuangteng Decoction 60 g/kg and 40 g/kg groups and sinomenine 28 mg/kg group. Immunohistochemical method showed that the expression of Bax protein was inhibited and the expression of Bcl-2 protein was risen in 60 g/kg,40 g/kg and 20 g/kg groups of Shuangteng Decoction and 28 mg/kg group of sinomenine. Western-blotting showed that the expressions of Caspase-1,Caspase-3 and CytC protein were inhibited in 60 g/kg,40 g/kg and 20 g/kg groups of Shuangteng Decoction and 28 mg/kg group of sinomenine respectively. Conclusion：Shuangteng Decoction could regulate apoptosis protein including Bax,Bcl-2,Caspase-1,Caspase-3,CytC protein expressions and inhibit the inflammation of collagen-induced arthritis rats aggravated by cold.