缓释bFGF促进缺血再灌注损伤皮瓣成活的实验研究

Improvement of slow-released bFGF on flap survival of ischemia-reperfusion injury

目的研究缓释bFGF对缺血再灌注损伤皮瓣的效应。方法采用酸性明胶水凝胶微球AGHMs为bFGF的载体，于10周龄20只雄性健康的Fisher大鼠背部制备对称性缺血再灌注损伤岛状皮瓣，即每只鼠2例皮瓣，共制备40例皮瓣；以CAM快速热成像仪检测皮瓣的缺血及再灌注；再灌注前注射AGHMs＋bFGF的溶液，分组为：对照A组（n=10例）为AGHM＋PBS溶液；实验组为bFGF＋AGHMs＋PBS溶液，其中的bFGF分别为20肛g（设为B组，n=10例）、50μg（设为C组，n=10例）和150μg（设为D组，n=10例）。手术后第7天对皮瓣成活率、微小血管成像、组织学及免疫组化检查进行评估。结果A、B、C、D组皮瓣的成活率分别为（52．27±10．20）％、（64．00±8．20）％、（64．30±10．10）％及（78．47±11．90）％。D组皮瓣成活率与其他3组相比，其差异有统计学意义（P〈0．01）。微小血管成像及组织学检查显示，D组的微小血管多于其他组；电镜检查显示，A、B、C组的组织损伤较重，D组的损伤轻微。免疫组化（VEGF，TGF—β1，TGF—β2，TGF-β3，bFGF）结果相近，即D组的阳性染色表面积与其他组相比，其差异有统计学意义（P〈0．01）；但A、B、C组之间相比，其差异无统计学意义（P〉0．1）。结论持续释放的bFGF可以上调局部组织VEGF及TGF的释放，促进新生血管的形成，从而提高缺血再灌注损伤皮瓣的成活率。

Objective To study the effect of slow-released bFGF on flap survival of ischemia-repeffusion injury. Methods AGHMs was used as the vector of bFGF. Island flaps （ n= 40） of ischemia-reperfusion injury were prepared symmetrically on the back of 10-week Fisher male rats （20）. The computer-aided design manufacture thermal imager was used to detect the ischemia and repeffusion of those flaps. AGHMs ＋ bFGF were injected before reperfusion. The flaps were divided randomly into the control group （ n = 10） which were treated with AGHMs ＋ bFGF only and the experimental group （ n = 30） which were treated with bFGF ＋ AGHMs ＋ PBS, and then the experimental group were sub-divided randomly into B, C and D groups （ n= 10 in each group）, which of each was administrated with 20 μg, 50 txg and 150 μg bFGF respectively. The flap survival, microangiography, histological and IHC examination were performed at 7 days postoperatively. Results The mean flap survival rates in the A, B, C and D groups were （52.27±10.20） %, （64.00±8.20） %, （64.30± 10. 10） % and （78.47±11.90） % respectively. Compared the D group with the other three groups, the difference had statistical significance （ P 〈0.01 ） ; the results of microangiography and histological examination revealed that the capillaries and small vessels in the D group were more than those in the others; tissue damage in the A, B and C groups by the EM was more severe than that in the D group; the results of VEGF, TGFβ1, TGFβ2, TGFβ3 and bFGF detected by IHC showed that the positive stained area in the D group was significantly higher than the others （ P〈0.01 ）, but the differences among A, B and C groups had no statistical significance （P〉0.1）. Conclusion The slow-released bFGF could regulate upward the secretion of VEGF and TGF, promoting the neovascular formation to improve the survival rate of the flaps of ischemia-reperfusion injury.