补肾活血方联合骨髓间充质干细胞-藻酸钙复合体修复关节软骨缺损研究

Research on repair of articular cartilage defect by Bushen Huoxue Formula combining bone marrow mesenchymal stem cell-calcium alginate compounds

目的:观察补肾活血方联合骨髓间充质干细胞(BMSCs)-藻酸钙复合体体治疗大鼠膝关节软骨缺损的疗效,探讨其作用机制。方法:48只大鼠全层软骨缺损造模随机分成4组:模型组、补肾活血组、BMSCs-藻酸钙组、补肾活血联合BMSCs-藻酸钙组。补血活血组术后8周每天给予补肾活血方灌胃;其余组术后每天给予等量0.9%氯化钠溶液灌胃。在第4、8周分批处死,收集并作相关检测。结果:形态学、病理组织学染色及Mankin评分结果表明,模型组软骨缺损损伤加重,伴有纤维组织增生。其余组缺损可见组织填充修复,尤以补肾活血联合BMSCs-藻酸钙组修复组织最佳。Real-time PCR结果证实,补肾活血联合BMSCs-藻酸钙组修复组织中Ⅱ型胶原、聚集蛋白聚糖的m RNA表达明显高于其余各组(P<0.01)。结论:补肾活血方和BMSCs-藻酸钙复合体单独使用对关节软骨缺损均具有一定的修复治疗作用,两者联合使用时通过增加Ⅱ型胶原和聚集蛋白聚糖合成,软骨修复效果达到最好。

Objective: To observe the effects of Bushen Huoxue Formula combining bone marrow mesenchymal stem cell(BMMSCs)-calcium alginate compounds in repairing articular cartilage defect and its underlying mechanisms. Methods: A total of 48 rats with full-thickness defects were randomly divided into four groups including model group, Bushen Huoxue Formula group, BMMSCs-calcium alginate compounds group, and Bushen Huoxue Formula combining BMMSCs-calcium alginate compounds group. The medication groups were continuously given Bushen Huoxue Formula for 8 weeks. Others were given equal volume of normal saline at same time. All rats were sacrificed in the 4th week and 8th week respectively for further detection. Results: The results of morphology, pathological histology and Mankin scores showed that the model group showed an aggravating damage on cartilage defect with fibrous hyperplasia. The defects were filled with white cartilage tissue in other groups, especially in the Bushen Huoxue Formula combining BMMSCs-calcium alginate compounds group. RT-PCR results showed that the m RNA expression of type Ⅱ collagen and aggrecan in Bushen Huoxue Formula combining BMMSCs-calcium alginate compounds groupwere increasedsignificantly compared to other groups(P<0.05). Conclusion: Both Bushen Huoxue Formula and BMMSCscalcium alginate compounds could repair cartilage defects. And, the combination of Bushen Huoxue Formula and BMMSCscalcium alginate compounds exerted a better effect on repairing the cartilage defects by increasing synthesis of type Ⅱcollagen and aggrecan.