肝素化羊脱细胞真皮基质对深Ⅱ度烧伤创面组织中血管内皮生长因子和层粘连蛋白的影响

目的研究肝素化羊脱细胞真皮基质(ADM)促进深Ⅱ度烧伤创面愈合的作用机制。 方法制作大鼠深Ⅱ度烧伤模型，随机分为碘伏纱布组、猪ADM组、羊ADM组和肝素化羊ADM组，每组40只。在伤后不同时间段，观察各组创面愈合率，同时在各个时间段处死大鼠8只，取创面组织标本做RT-qPCR和蛋白质印迹法检测各组创面组织中血管内皮生长因子(VEGF)和层粘连蛋白(LN)的mRNA与蛋白表达情况。对数据行单因素方差分析和t检验。 结果肝素化羊ADM组深Ⅱ度创面的愈合率优于碘伏纱布组。RT-qPCR检测发现，VEGF的mRNA在各组伤后3 d的创面组织中表达较对照组高，各组间的比较发现，伤后7 d，肝素化羊ADM组的VEGF的mRNA表达量明显高于碘伏纱布组(t＝80.83，P<0.05)，羊ADM组和猪ADM组在伤后7 d也较碘伏纱布组的表达量高(t＝60.80、53.42，P值均小于0.05)；蛋白质印迹法检测肝素化羊ADM组伤后3 d时VEGF蛋白表达量较多，随后便逐渐减少，伤后21 d时蛋白表达较少。伤后3 d,碘伏纱布组LN在创面的表达明显下降，在之后的治疗过程中，LN的表达逐渐恢复，伤后14 d的表达量明显高于伤后3 d(t＝19.5，P<0.001)，伤后21、28 d的表达量与伤后14 d相差不明显。蛋白质印迹法检测发现肝素化羊ADM组LN在伤后3 d便开始出现表达量逐渐增多，伤后21 d时出现蛋白表达较多。 结论肝素化羊ADM能够保护创面及促进创面组织中VEGF和LN的表达，进而促进创面愈合。

Effect of heparinized sheep acellular dermal matrix on vascular endothelial growth factor and laminin in deep partial-thickness burn wound tissue

ObjectiveTo investigate the mechanism of heparin-selective sheep acellular dermal matrix in healing wound healing of deep partial-thickness burn. MethodsRat deep partial-thickness models were made and randomly divided into iodophor gauze group, pig acellular dermal matrix (ADM) group, sheep ADM group and heparinized sheep ADM group, each group of 40 rats. The wound healing rate in the group was also sacrificed at various time points. Eight rats were killed at different times. The wound tissue samples were taken for RT-qPCR and Western-blotting to detect vascular endothelial growth factor (VEGF) and laminin(LN) in the wound tissues of each group. Data were processed with one-way analysis of variance and t test. ResultsThe wound healing rate of the heparinized sheep ADM group in the treatment was better than that of the iodophor gauze group. RT-qPCR analysis showed that the expression of VEGF in the wound tissue at 3 days post-operation after scald in each group was higher than in the control group. The comparison between the groups showed that the expression of VEGF in heparinized sheep ADM group was significantly higher than that in iodophor group (t=80.83, P<0.05). The sheep ADM group and the pig ADM group were also higher than the iodophor group at 7 days post-operation (t=60.80, 53.42; with P values below 0.05). Western-blotting was used to detect the VEGF wound in the heparinized sheep ADM group. At 3 days post-operation, the expression was more, and then gradually decreased. At 21 days post-operation, the protein expression was less. At 3 days post-operation, the expression of LN in the iodophor group decreased significantly. During the subsequent treatment, the expression of LN gradually recovered, and the expression level at 14 days post-operation was significantly higher than that at 3 days post-operation group (t=19.5, P<0.001). The expression levels of 21 days post-operation and 28 days post-operation were not significantly different from those of 14 days post-operation. The Western-blotting showed that the LN of the heparinized sheep ADM group began to increase gradually at 3 days post-operation after the wound surface, and the protein expression appeared more on the 21 days post-operation. ConclusionHeparinized sheep ADM can protect the wound surface and promote the expression of VEGF and LN in the wound tissue, which in turn promotes wound healing.