MEBT/MEBO对大鼠慢性难愈合创面组织中MMP-2、CXCL1、GM-CSF蛋白表达的影响

Effects of MEBT/MEBO on the expressions of MMP-2,CXCL1 and GM-CSF proteins in chronic refractory wounds in rats

目的观察湿润暴露疗法/湿润烧伤膏(MEBT/MEBO)对大鼠慢性难愈合创面组织中基质金属蛋白酶2(MMP-2)、趋化因子配体1(CXCL1)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)蛋白表达的影响。方法将72只SD大鼠随机分为急性皮损组、慢性创面组、贝复新组、美宝组各18只,除急性皮损组外,其余组皮下注射氢化可的松建立慢性难愈合创面,注射完成后用生理盐水清理创面,贝复新组创面敷贝复新凝胶,美宝组创面敷美宝烧伤膏,均每日上下午各换药1次。观察各组大鼠伤后第3,7,14天创面情况并取创面肉芽组织,Masson染色观察创面中胶原纤维生成情况,ELISA法检测肉芽组织中MMP-2含量,免疫组化法检测创面组织中CXCL1、GM-CSF蛋白表达情况。结果美宝组大鼠创面愈合速度快于慢性创面组和贝复新组。Masson染色显示各组创面组织中胶原纤维含量随伤后时间的延长逐渐增加;伤后第3,7天,美宝组胶原纤维含量均明显高于慢性创面组和贝复新组(P均<0.05);伤后第14天,各组胶原纤维含量比较差异均无统计学意义(P均>0.05)。创面组织中MMP-2含量,贝复新组随伤后时间延长而增加(P<0.05),美宝组随伤后时间延长而减少(P<0.05);伤后第3天,美宝组创面组织中MMP-2含量明显高于慢性创面组(P<0.05);伤后第7天,美宝组和贝复新组创面组织中MMP-2含量下降但与慢性创面组比较差异均无统计学意义(P均>0.05);伤后第14天,美宝组创面组织中MMP-2含量明显低于慢性创面组和贝复新组(P均<0.05)。贝复新组伤后创面组织中CXCL1表达平均光密度随伤后时间延长降低,伤后第14天明显低于伤后第3天(P<0.05);伤后第3,7,14天创面组织中GM-CSF表达平均光密度比较差异均无统计学意义(P均>0.05)。美宝组伤后第3,7,14天创面组织中CXCL1表达平均光密度比较差异均无统计学意义(P均>0.05);伤后第7,14天创面组织中GM-CSF表达平均光密度均明显低于伤后第3天(P均<0.05),且伤后第14天明显低于伤后第7天(P<0.05)。慢性创面组、贝复新组、美宝组伤后第3,7,14天创面组织中CXCL1表达平均光密度和伤后第7天创面组织中GM-CSF表达平均光密度比较差异均无统计学意义(P均>0.05)。伤后第3天,美宝组创面组织中GM-CSF表达平均光密度与慢性创面组比较差异无统计学意义(P>0.05),但明显高于贝复新组(P<0.05);伤后第14天,美宝组创面组织中GM-CSF表达平均光密度明显低于慢性创面组(P<0.05)。结论MEBT/MEBO可能通过促进胶原纤维的生成,调节MMP-2及GM-CSF、CXCL1蛋白的表达,从而加快慢性难愈合创面组织的修复。

Objective It is to observe the effect of moist exposed burn therapy(MEBT)/moist exposed burn ointment(MEBO)on the expression of matrix metalloproteinase-2(MMP-2),chemokine ligand 1(CXCL1),granulocyte-macrophage colony-stimulating factor(GM-CSF)in chronic refractory wound tissue in rats.Methods Seventy-two SD rats were randomly divided into 4 groups:acute skin lesion group,chronic wound group,Beifuxin group,and Meibao group,with 18 rats in each group.Except for the acute skin lesion group,the other groups were subcutaneously injected with hydrocortisone to establish chronic refractory wound models.After the injection was completed,the wound was cleaned with normal saline,Befexin gel was applied to the wound in the Befexin group,and Meibao burn cream was applied to the wound in the Meibao group,and the dressing was changed once in the morning and in the afternoon.The wounds of the rats in each group were observed and their granulation tissue was collected at 3,7,and 14 days after injury,the production of collagen fiber in the trauma was observed by Masson staining,the content of MMP-2 in the granulation tissue was detected by ELISA,and the expression of CXCL1 and GM-CSF protein in the trauma tissue was detected by immunohistochemistry.Results The wound healing speed of the rats in the Meibao group was faster than that in the model group and Beifuxin group.Masson staining showed that the content of collagen fibers in the wound tissue of each group was gradually increased with time expanded after injury;on the 3rd and 7th day after injury,the expression of collagen fibers in the Meibao group was significantly higher than that in the chronic wound group and Beifuxin group(P<0.05),but there was no significant difference in the expression among each group on the 14th day(all P>0.05).The content of MMP-2 in traumatic tissue was increased with time expanded after injury in the Befexin group(P<0.05)and was decreased with time expanded after injury in the Meibao group(P<0.05);on the 3rd day after injury,the content of MMP-2 in traumatic tissue of the Meibao group was significantly higher than that of the chronic wound group(P<0.05);on the 7th day after injury,the content of MMP-2 in traumatic tissue of the Meibao group and Beifuxin group were decreased,but the difference was not statistically significant compared with the chronic wound group(P>0.05);on the 14th day after injury,the content of MMP-2 in traumatic tissue of the Meibao group was significantly lower than that of the chronic wound group and Beifuxin group(P<0.05).The mean optical density of CXCL1 expression in traumatic tissue of the Befuxin group was decreased with time expanded after injury,and it was significantly lower on the 3rd day after injury than that on the 14th day after injury(P<0.05);the differences in the mean optical density of GM-CSF expression in traumatic tissue on the 3rd,7th,and 14 th days after injury were not statistically significant(all P>0.05).The differences in the mean optical density of CXCL1 expression in traumatic tissues on the 3rd,7th and 14th days after injury in the Meibao group were not statistically significant(all P>0.05);the mean optical density of GM-CSF expression in traumatic tissues on the 7th and 14th days after injury was significantly lower than that on the 3rd day(P<0.05),and that on the 14th day was significantly lower than that on the 7th day(P<0.05).The differences in the mean optical density of CXCL1 expression in trauma tissue on the 3rd,7th and 14th days after injury,and the mean optical density of GM-CSF expression in trauma tissue on the 7 th day were not statistically significant among the chronic wound group,Befuxin group and Meibao group(all P>0.05).On the 3rd day after injury,the difference in the mean optical density of GM-CSF expression in trauma tissue of the Meibao group was not statistically significant compared with that of the chronic wound group(P>0.05),but it was significantly higher than that of the Befuxin group(P<0.05);on the 14th day after injury,the mean optical density of GM-CSF expression in trauma tissue of the Meibao group was significantly lower than that of the chronic wound group(P<0.05).Conclusion MEBT/MEBO may accelerate the repair of chronic refractory wound tissue via promoting the formation of collagen fibers,regulating the changes of MMP-2 content and the expression of GM-CSF and CXCL1 proteins.