CAJ | 학술논문

目的探讨巨噬细胞(macrophage)及粒细胞巨噬细胞集落刺激因子(GM-CSF)促进大鼠腹壁动脉穿支(DEP)皮瓣模型皮瓣存活的作用及分子机制.方法建立大鼠DEP皮瓣模型,分别给与大鼠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,测算皮瓣成活面积、微血管密度(MVD),天狼腥红染色检测胶原沉积,并利用逆转录-聚合酶链反应(RT-PCR)法检测I型胶原(Collagen Ⅰ)、Ⅲ型胶原(Collagen Ⅲ)、基质金属蛋白酶-2(MMP2)及基质金属蛋白酶诱导因子(EMMPRIN)、血管内皮生长因子(VEGF)及其受体(VEGFR)的基因表达.结果皮瓣成活率Ⅰ组(53.08±8.76)%和Ⅱ组(47.95±4.92)%均高于对照组(43.28±5.27)%而低于Ⅲ组(61.68±6.60)%,差异有统计学意义(P＜0.05).胶原含量Ⅰ、Ⅲ组(16.34±2.47)%、(19.41±2.01)%均高于对照组(13.19±2.91)%,差异有统计学意义(P均＜0.01);Ⅱ组(12.50±2.66)%稍低于对照组,差异无统计学意义(P＞0.05).MVD Ⅰ～Ⅲ组(24.82±4.18、24.30±3.02、29.82±4.74)均高于Ⅳ组(21.37±2.65),差异有统计学意义,其中Ⅲ组MVD较其他两组MVD更高.Ⅰ组皮瓣Collagen Ⅰ、EMMPRIN、MMP2和VEGFR基因表达高于对照组(P＜0.05);Ⅱ组皮瓣EMMPRIN基因表达高于对照组(P＜0.05);Ⅲ组皮瓣Collagen Ⅰ、Collagen Ⅲ、EMMPRIN、MMP2和VEGFR基因表达均高于对照组(P＜0.05).各实验组较对照组VEGF表达差异无统计学意义(P＞0.05).结论重组大鼠GM-CSF和过继巨噬细胞通过促进皮瓣内胶原合成(Collagen Ⅰ、Collagen Ⅲ)及分解(EMMPRIN、MMP2)相关基因的表达,同时促进血管生成相关基因(VEGFR)的表达,促进了皮瓣胶原重塑和血管生成,最终促进了大鼠DEP皮瓣的成活.
목적 탐토거서세포(macrophage)급립세포거서세포집락자격인자(GM-CSF)촉진대서복벽동맥천지(DEP)피판모형피판존활적작용급분자궤제.방법 건립대서DEP피판모형,분별급여대서GM-CSF(Ⅰ조)、복강거서세포(Ⅱ조)、GM-CSF연합거서세포(Ⅲ조)급생리염수(Ⅳ조).술후제7천취피판,측산피판성활면적、미혈관밀도(MVD),천랑성홍염색검측효원침적,병이용역전록-취합매련반응(RT-PCR)법검측I형효원(Collagen Ⅰ)、Ⅲ형효원(Collagen Ⅲ)、기질금속단백매-2(MMP2)급기질금속단백매유도인자(EMMPRIN)、혈관내피생장인자(VEGF)급기수체(VEGFR)적기인표체.결과 피판성활솔Ⅰ조(53.08±8.76)%화Ⅱ조(47.95±4.92)%균고우대조조(43.28±5.27)%이저우Ⅲ조(61.68±6.60)%,차이유통계학의의(P＜0.05).효원함량Ⅰ、Ⅲ조(16.34±2.47)%、(19.41±2.01)%균고우대조조(13.19±2.91)%,차이유통계학의의(P균＜0.01);Ⅱ조(12.50±2.66)%초저우대조조,차이무통계학의의(P＞0.05).MVD Ⅰ～Ⅲ조(24.82±4.18、24.30±3.02、29.82±4.74)균고우Ⅳ조(21.37±2.65),차이유통계학의의,기중Ⅲ조MVD교기타량조MVD경고.Ⅰ조피판Collagen Ⅰ、EMMPRIN、MMP2화VEGFR기인표체고우대조조(P＜0.05);Ⅱ조피판EMMPRIN기인표체고우대조조(P＜0.05);Ⅲ조피판Collagen Ⅰ、Collagen Ⅲ、EMMPRIN、MMP2화VEGFR기인표체균고우대조조(P＜0.05).각실험조교대조조VEGF표체차이무통계학의의(P＞0.05).결론 중조대서GM-CSF화과계거서세포통과촉진피판내효원합성(Collagen Ⅰ、Collagen Ⅲ)급분해(EMMPRIN、MMP2)상관기인적표체,동시촉진혈관생성상관기인(VEGFR)적표체,촉진료피판효원중소화혈관생성,최종촉진료대서DEP피판적성활.
Objective To study the promotive effect of macrophage and granulocyte macrophage colony-stimulating factor (GM-CSF) on the survival of deep epigastric perforator (DEP) in rats and the mechanism. Methods A stable animal model of DEP flap in Sprague-Dawley rats was established for human deep inferior epigastric perforator flap breast reconstruction. The flaps were treated respectively by the method of subcutaneous injection of recombinant rat GM-CSF ( group Ⅰ ) , rat peritoneal macrophages (group Ⅱ ), GM-CSF combined with macrophages ( group Ⅲ ), and saline as parallel negative control ( group Ⅳ). The rats were sacrified and flap specimens were harvested on the 7th day after operation. The survival rate of flaps and micro-vesscle density (MVD) were measured. Sirius Red staining was done to analyze collagen deposition, and the mRNA expression of collagen I , collagen Ⅲ, extracellular matrix matalloproteinase inducer (EMMPRIN) , matrix metalloproteinase 2 ( MMP-2), vascular endothelial growth factor (VEGF) and VEGFR was detected by reverse transcription-polymerase chain reaction (RT-PCR).Results Survival rate of the flaps in both group Ⅰ [(53.08 ±8. 76)%] and group Ⅱ [(47.95 ±4. 92)%] was significantly higher than in control group [(43.28 ±5. 27)%], but significantly lower than in group Ⅲ [(61.68±6.60)%]. Collagen deposition in groups Ⅰ [(16. 34 ±2.47)%] and Ⅲ [(19.41 ±2.01)%] was significantly higher than in control group [(13. 19 ±2.91)%]. But collagen deposition in group Ⅱ [(12. 50 ± 2. 66)%] was lower than in control group, though there was no significant difference (P＞0.05). MVD in groups Ⅰ,Ⅱ and Ⅲ (24. 82 ±4. 18, 24.30 ±3.02, 29. 82 ±4.74 respectively) was significantly higher than in control group (21. 37 ± 2. 65 ) , and that in group Ⅲ was significantly higher than the others. The results of RT-PCR showed collagen Ⅰ, EMMPRIN, MMP2 and VEGFR expression levels were significantly higher in group Ⅰ than in control group ( P ＜ 0. 05 ) ; EMMPRIN expression was significantly higher in group Ⅱ than in control group (P ＜0.05); Collagen Ⅰ, Collagen Ⅲ, EMMPRIN, MMP2 and VEGFR expression levels in group Ⅲ were significantly higher than in control group ( P ＜ 0. 05 ). There was no significant difference between groups ⅠⅢ and control group (P＞0.05). Conclusion By promoting the expression of collagen synthesis (collagen Ⅰ, collagen Ⅲ) and degradation (EMMPRIN, MMP2) related genes,and expression of angiogenesis related gene (VEGFR), recombinant rat GM-CSF and macrophages enhanced collagen remodeling and angiogenesis, and ultimately promoted the survival of DEP flaps in rats.