巨噬细胞及粒巨细胞-集落刺激因子对大鼠腹壁皮瓣成活的影响

The effect of macrophage and granulocyte macrophage colony-stimulating factor on the survival of rat abdominal wall flap

目的研究巨噬细胞及粒巨细胞-集落刺激因子（GN—CSF）对大鼠腹壁动脉穿支（deep epigastric perforator，DEP）皮瓣模型的影响。方法建立SD大鼠DEP皮瓣模型，分别给予大鼠GM—CSF（I组）、腹腔巨噬细胞（Ⅱ组）、GM—CSF联合巨噬细胞（Ⅲ组）及生理盐水（Ⅳ组）。术后第7天取皮瓣检测成活面积、组织学观察、微血管密度（MVD）、皮瓣内胶原含量。结果皮瓣成活率I组（53．08％±8．76％）和Ⅱ组（47．95％4-4．92％）间无差异，均高于Ⅳ组（43．28％±5．27％）而低于Ⅲ组（61．68％±6．60％），P〈0．05。皮瓣MVD1组（24．82±4．18）和Ⅱ组（24．30±3．02）间差异无统计学意义，均显著高于Ⅳ组（21．37±2．65），低于Ⅲ组（29．82±4．74）。胶原含量I组（17．25％±2．85％）高于Ⅳ组（14．41％±2．89％），P〈0．05。II组（12．69％±3．55％）稍低于Ⅳ组。Ⅲ组（20．31％±3．01％）较I组显著增高，P〈0．05。结论重组大鼠GM—CSF和巨噬细胞均能够促进大鼠DEP皮瓣成活，联合应用可发挥协同作用促进皮瓣存活、血管生成以及胶原沉积。

Objective To study the effect of macrophage, its stimulating factor, granulocyte macrophage colony-stimulating factor （GM-CSF）, the combination of GM-CSF and macrophage on the survival of rat deep epigastric perforator flap （DEP）. Methods The stable animal model of DEP flap in Sprague-Dawley rat mimicing human deep inferior epigastric perforator flap in breast reconstruction was established. The rats were treated with subcutaneous injection of recombined rat GM-CSF or rat peritoneal macrophages, respectively, or combination of GM-CSF/ Macrophages. Normal saline was used as parallel negative control. The rats were sacrificed and flap specimens were harvested on day 7 after operation, the flaps survival area were measured by the method of rubbings and the survival proportion of flaps were calculated, Von Will brand factor were detected by immunohistocbemistry and microvessel density （MVD） , and were calculated with microscopic study, and collagen were stained and quantified by Masson staining. Results Survival proportion of flaps in group GM-CSF （ 53.08% ± 8.76% ） was not different with that in macrophages group （47. 95% ± 4. 92% ）, and both of these two groups were significantly higher than parallel negative control group （43.28% ± 5.27% ） but significantly lower than combination GM-CSF/ macrophages group （61.68% ± 6. 60% ）. For MVD, flap in GM-CSF group （24.82 ± 4. 18 ） was not significantly different with macrophages group （ 24. 30 ± 3.02 ）, and both of these two groups were significantly higher than group parallel negative control （21.37 ±2. 65 ） but significantly lower than combination GM-CSF/macrophages group （ 29. 82 ± 4. 74 ）. Collagen deposition in the flaps in GM-CSF group （17.25% ±2.85% ） were significantly higher than parallel negative control group （14.41% ± 2. 89% ）, macrophages group （ 12. 69% ±3.55% ） were lower than parallel negative control group but there was no significant difference. That in combination GM-CSF/macrophages group （20. 31% ± 3. 01% ） was significantly higher than GM-CSF group （P 〈 0. 05 ）. Conclusion Treatment with rat GM-CSF or macrophage can significantly promote the survival of the flaps. Combined application of GM-CSF and macrophage could synergetically promote the survival of the flaps, the vasculogenesis and the collagen deposition.