脐带间充质干细胞上清冻干粉对大耳兔皮肤缺损的治疗作用

Effect of supernatant of umbilical cord mesenchymal stem cells on skin defect in flapeared rabbits

目的探讨脐带间充质干细胞(MSCs)上清提取物治疗大耳兔耳部皮肤缺损的作用。方法将12只日本大耳兔随机分为对照组、单纯培养基冻干粉治疗组和MSCs上清冻干粉治疗组,每组4只。3组大耳兔在耳部制作皮肤缺损成功后分别给予生理盐水、单纯培养基冻干粉和MSCs上清液冻干粉治疗。模型制作后第3、5、7天检测血清中肿瘤坏死因子-α(TNF-α)水平;模型制作第7天检测3组大耳兔损伤区域周围组织内TNF-α和周围血管内皮生长因子(VEGF)水平。结果由脐带培养的MSCs形态呈梭形、圆钝,漩涡状生长,较为饱满。皮肤缺损后第3、5、7天,MSCs上清冻干粉治疗组大耳兔血清中TNF-α水平均低于对照组和单纯培养基冻干粉治疗组(P<0.05),且单纯培养基冻干粉治疗组高于对照组(P<0.05),第7天时皮肤缺损周围组织中TNF-αmRNA及蛋白表达与血清一致;MSCs上清冻干粉治疗组大耳兔损伤区周围组织内VEGF mRNA及蛋白表达均高于对照组和单纯培养基冻干粉治疗组(P<0.05);MSCs上清冻干粉治疗组大耳兔损伤区周围组织内TNF-αmRNA及蛋白表达均低于对照组和单纯培养基冻干粉治疗组(P<0.05)。结论皮肤缺损大耳兔行MSCs上清冻干粉治疗后可有效减轻炎症反应,改善损伤区域血管重建,有益于缺损皮肤恢复。

Objective To study the effect of supernatant of umbilical cord mesenchymal stem cells( MSCs) on skin defect in flapeared rabbits. Methods Twelve Japanese flapeared rabbits were randomly divided into control group,culture medium treatment group and MSCs supernatant treatment group,four rabbits in each group. The rabbits in control group were treated with normal saline,the rabbits in culture medium treatment group were treated with culture medium lyophilized powder,and the rabbits in MSCs supernatant treatment group were treated with MSCs supernatant lyophilized powder. The level of serum tumor necrosis factor-α( TNF-α) were detected at the 3rd,5thand 7thafter model building; the TNF-α and vascular endothelial growth factor( VEGF) in the tissues of skin defect of rabbits in the three groups were detected at 7thafter model building. Results The shape of MSCs from umbilical cord was blunt clostridial form and swirl like growth. The level of serum TNF-α of rabbits in MSCs supernatant treatment group was significantly lower than that in control group and culture medium treatment group( P <0. 05),and the level of serum TNF-α in culture medium treatment group was significantly higher than that in control group at the 3rd,5thand 7thafter model building( P < 0. 05). The expressions of VEGF mRNA and protein in the tissues of skin defect of rabbits in MSCs supernatant treatment group were significantly higher than those in control group and culture medium treatment group( P < 0. 05). The expressions of TNF-α mRNA and protein of rabbits in MSCs supernatant treatment group were significantly lower than those in control group and culture medium treatment group( P < 0. 05). Conclusion The supernatant of umbilical cord MSCs can effectively reduce inflammatory reaction,improve vascular remodeling in injured area,which is helpful to the recovery of the skin defect in rabbits.