柚皮苷联合骨髓间充质干细胞结合脱细胞真皮基质对软骨损伤的修复作用及机制

Repair effect and mechanism of naringin combined with bone marrow mesenchymal stem cells and acellular dermal matrix on cartilage injury

目的探讨柚皮苷联合骨髓间充质干细胞结合脱细胞真皮基质对兔膝关节软骨损伤的修复作用及其机制。方法将12只新西兰大耳白兔随机分为模型组、细胞组、柚皮苷联合细胞组,每组4只。关节软骨缺损造模后,模型组给予去离子水灌胃;细胞组在关节缺损处植入骨髓间充质干细胞结合脱细胞真皮基质,给予去离子水灌胃;柚皮苷联合细胞组在关节缺损处植入骨髓间充质干细胞结合脱细胞真皮基质,给予柚皮苷汤灌胃。连续灌胃12周后处死各组白兔,采用O’Driscoll评分系统、Mankin’s评分系统评估软骨修复和缺损情况,并对关节软骨缺损区域行病理学染色及免疫组化染色观察。结果细胞组和柚皮苷联合细胞组O’Driscoll评分均明显高于模型组(P均<0.05),且柚皮苷联合细胞组明显高于细胞组(P<0.05);细胞组和柚皮苷联合细胞组Mankin’s评分均明显低于模型组(P均<0.05),且柚皮苷联合细胞组明显低于细胞组(P<0.05)。苏木精-伊红染色、甲苯胺蓝染色、番红-O染色显示,细胞组和柚皮苷联合细胞组可见明显软骨修复改变,柚皮苷联合细胞组改善更明显;细胞组在关节缺损修复处稍有转化生长因子-β2(TGF-β2)、转化生长因子-β3(TGF-β3)分泌;柚皮苷联合细胞组可见明显分泌的TGF-β2、TGF-β3,存在很少量的X型胶原染色。结论柚皮苷联合骨髓间充质干细胞结合脱细胞真皮基质对于兔膝关节软骨损伤有修复作用,该修复效果的获得可能与柚皮苷加速骨髓间充质干细胞软骨方向分化及刺激TGF-β2、TGF-β3分泌有关。

Objective It is to explore the repair effect and mechanism of naringin combined with bone marrow mesenchymal stem cells and acellular dermal matrix on cartilage injury.Methods 12 New Zealand big-eared rabbits were randomly divided into 3 groups:model group,cell group,naringin combined cell group(combination group),4 rabbits in each group.After the articular cartilage defect was modeled,the model group was given deionized water by gavage;the cell group was given bone marrow mesenchymal stem cells combined with acellular dermal matrix in the joint defect and deionized water by gavage;the combination group was given bone marrow mesenchymal stem cells combined with acellular dermal matrix in the joint defect and naringin decoction by gavage.After 12 weeks of continuous gastric administration,the rabbits in each group were sacrificed.The O’Driscoll scoring system and Mankin’s scoring system were used to evaluate cartilage repair and defect,and the articular cartilage defect area was observed by pathological staining and immunohistochemical staining.Results The O’Driscoll scores of the cell group and the combination group were significantly higher than those of the model group(P<0.05),and the scores in the combination group were significantly higher than those in the cell group(P<0.05);the Mankin’s scores of the cell group and the combination group were significantly lower than those of the model group(P<0.05),and the scores in the combination group were significantly lower than those in the cell group(P<0.05).Hematoxylin-eosin staining,toluidine blue staining,and safranin-O staining showed that obvious cartilage repair changes were found in the cell group and combination group,and the improvements in the combination group were more significant.There was a slight secretion of transforming growth factor-β2(TGF-β2)and transforming growth factor-β3(TGF-β3)at the repair site of joint defects in the cell group;TGF-β2 and TGF-β3 were obviously secreted in the combination group,with a small amount Type X collagen staining.Conclusion Naringin combined with bone marrow mesenchymal stem cells and acellular dermal matrix can repair cartilage damage in rabbit knee joint.The repair effect may be related to the acceleration of chondrogenic differentiation of bone marrow mesenchymal stem cells and the stimulation of TGF-β2,TGF-β3 secretion by naringin.