中成药无敌丹对骨关节炎软骨细胞活力和软骨细胞凋亡的影响

Effect of Wudi Dan on vitality and apoptosis of cartilage chondrocytes after osteoarthritis

背景:研究发现,无敌丹能够抑制病变关节局部的炎症反应,保护关节软骨。目的:验证无敌丹对家兔软骨细胞活力和软骨细胞凋亡的影响及对家兔骨关节炎病变的治疗效果。方法:用含无敌丹的血清培养基培养大鼠软骨细胞,通过与溶剂对照组对比,观察无敌丹对软骨细胞活力及凋亡的影响;采用改良Hulth法构建兔膝骨关节炎模型,无敌丹给药组给予无敌丹;溶剂对照组给予生理盐水,2次/d,连续给药4周。观察无敌丹对兔膝骨关节炎的治疗作用;镜下观察关节局部的软骨细胞情况;用免疫组织化学的方法检测软骨中白细胞介素1、基质金属蛋白酶3的水平。结果与结论:无敌丹血清培养的软骨细胞凋亡率显著低于对照组;兔膝关节病理检查显示溶剂对照组软骨破坏程度较高,无敌丹组软骨破坏程度低;免疫组织化学染色显示无敌丹给药组胞质和胞外基质着色浅,阳性细胞数量少,白细胞介素1表达低;无敌丹给药组阳性表达的细胞数量少,着色浅,基质金属蛋白酶3表达被抑制。结果表明,无敌丹能够有效保护家兔骨关节炎病变部位关节软骨,减少炎症反应,对于骨关节炎有很好的治疗作用,其机制可能与抑制软骨细胞凋亡、减少细胞因子的生成及抑制基质金属蛋白酶的蛋白表达有关。

BACKGROUND;Studies have found that Wudi Dan can suppress local inflammatory response of the lesioned joints to protect the articular cartilage.OBJECTIVE;To verify the effect of Wudi Dan on chondrocyte viability and apoptosis as well as the therapeutic effect on osteoarthritis.METHODS;Rat chondrocytes were cultured in serum medium containing Wudi Dan,and the effects of Wudi Dan on cell viability and apoptosis were observed by comparison with the control group.Rabbit model of knee osteoarthritis was constructed using modified Hulth method.Rabbit models were divided into two groups;Wudi Dan group treated with Wudi Dan and control group treated with normal saline,twice a day,consecutively for 4 weeks.Therapeutic effect of Wudi Dan on knee osteoarthritis was observed; cell viability and apoptosis were observed under microscope; the levels of interleukin-1 and matrix metalloproteinase-3 were determined using immunohistochemical method.RESULTS AND CONCLUSION;The apoptotic rate of chondrocytes was significantly lower in the Wudi Dan group than the control group.Pathological findings of the rabbit knee joints showed that the control group had more severe damage to the articular cartilage than the Wudi Dan group.Immunohistochemical staining revealed that in the Wudi Dan group,the cytoplasm and extracellular matrix were colored light and there were a small number of positive cells as well as low expression of interleukin-1 and matrix metalloproteinase-3.The results suggest that Wudi Dan can effectively protect against articular cartilage lesions,reduce inflammation,and have agood therapeutic effect on osteoarthritis.Its mechanism may be related to inhibition of chondrocyte apoptosis,reduction of cytokine production and inhibition of protein expression of matrix metalloproteinase.