骨髓间充质干细胞通过增加血管生成和血管内皮生长因子表达影响大鼠随意型皮瓣存活实验研究

Experimental study of BMSCs on survival of rat random flaps by increasing angiogenesis and VEGF expression

目的:探讨骨髓间充质干细胞(BMSCs)通过增加血管生成和血管内皮生长因子(VEGF)表达影响大鼠随意型皮瓣存活的效果。方法:将随意型皮瓣模型大鼠随机平分为模型组、小剂量BMSCs组与大剂量BMSCs组,每组16只。模型组注射0.4 ml磷酸盐缓冲液,小剂量BMSCs组与大剂量BMSCs组于相同部位注射等体积BMSCs完全培养液(细胞浓度分别为2×10^(8)个/L和20×10^(8)个/L)。比较三组大鼠皮瓣存活率及超氧化物歧化酶(SOD)、丙二醛(MDA)、基质金属蛋白酶9(MMP-9)、基质细胞衍生因子-1(SDF-1)、VEGF蛋白相对表达水平。结果:小剂量BMSCs组与大剂量BMSCs组治疗第7、14、28天的皮瓣存活率高于模型组,且大剂量BMSCs组皮瓣存活率高于小剂量BMSCs组(均P<0.05)。小剂量BMSCs组与大剂量BMSCs组治疗第7、14、28天的皮瓣SOD水平高于模型组,MDA水平低于模型组(均P<0.05)。小剂量BMSCs组治疗第7、14、28天的皮瓣SOD水平低于大剂量BMSCs组,MDA水平高于大剂量BMSC s组(均P<0.05)。小剂量BMSCs组与大剂量BMSCs组治疗第7、14、28天的血清MMP-9与SDF-1水平高于模型组,且大剂量BMSCs组MMP-9与SDF-1水平高于小剂量BMSCs组(均P<0.05)。小剂量BMSCs组与大剂量BMSCs组治疗第28、56天的皮瓣VEGF蛋白相对表达量高于模型组,而且大剂量BMSCs组VEGF蛋白相对表达量高于小剂量BMSCs组(均P<0.05)。结论:BMSCs可以提高大鼠血清MMP-9与SDF-1水平,增加皮瓣SOD水平,降低MDA水平,有利于提高VEGF的表达,从而增加随意型皮瓣存活率。

Objective:To investigate the effect of bone marrow mesenchymal stem cells(BMSCs)on the survival of rat random flaps by increasing angiogenesis and vascular endothelial growth factor(VEGF)expression.Methods:Random flap model rats were randomly divided into model group,low-dose BMSCs group and high-dose BMSCs group,with 16 rats in each group.The model group was injected with 0.4 ml phosphate buffer,and the low-dose BMSCs group and the high-dose BMSCs group were injected with the same volume of BMSCs complete culture medium at the same site(the cell concentration was 2×10^(8)/L and 20×10^(8)/L).The flap survival rate and the levels of superoxide dismutase(SOD),malondialdehyde(MDA),matrix metalloproteinase-9(MMP-9),stromal cell-derived factor-1(SDF-1)and the relative expression level of VEGF protein were compared among the three groups.Results:The flap survival rate of low-dose BMSCs group and high-dose BMSCs group on the 7th,14th and 28th day of treatment was higher than that of model group,and the flap survival rate of high-dose BMSCs group was higher than that of low-dose BMSCs group(all P<0.05).The SOD level of flap in low-dose BMSCs group and high-dose BMSCs group on the 7th,14th and 28th day of treatment was higher than that in model group,and the MDA level was lower than that in model group(all P<0.05).On the 7th,14th and 28th day after treatment,the SOD level of flap in low-dose BMSCs group was lower than that in high-dose BMSCs group,and the MDA level was higher than that in high-dose BMSCs group(all P<0.05).On the 7th,14th and 28th day after treatment,the levels of serum MMP-9 and SDF-1 in low-dose BMSCs group and high-dose BMSCs group were higher than those in model group,and the levels of MMP-9 and SDF-1 in high-dose BMSCs group were higher than those in low-dose BMSCs group(all P<0.05).The relative expression level of VEGF protein in the flap of low-dose BMSCs group and high-dose BMSCs group on the 28th and 56th day of treatment was higher than that of the model group,and the relative expression level of VEGF protein in high-dose BMSCs group was higher than that in low-dose BMSCs group(all P<0.05).Conclusion:BMSCs can increase the levels of serum MMP-9 and SDF-1,increase the level of SOD and reduce the level of MDA,which is conducive to improve the expression of VEGF and increase the survival rate of random flap.