重组腺病毒介导的血管内皮生长因子165基因增强大鼠横形腹直肌肌皮瓣活力的研究

STUDIES ON ENHANCEMENT OF TRANSVERSE RECTUS ABDOMINIS MUSCULOCUTANEOUS FLAP SURVIVAL BY RECOMBINANT ADENOVIRUS MEDIATED VASCULAR ENDOTHELIAL GRWOTH FACTOR 165 GENE

目的探讨重组腺病毒介导的血管内皮生长因子165(adenovirusmediatedvascularendothelialgrowthfactor165,Ad-VEGF165)基因,增强大鼠横形腹直肌肌皮瓣(transverserectusabdominismusculocutaneousflap,TRAM)局部缺血区域活力的基因治疗。方法SD大鼠30只,制备30mm×80mm以下腹部血管作为血管蒂的右侧矩形TRAM皮瓣,随机平均分成5组,分别在大鼠TRAM皮瓣皮下、皮下+腹直肌及腹直肌局部注射Ad-VEGF165(1～3组:治疗组),同时在TRAM皮瓣皮下+腹直肌局部注射重组腺病毒载体介导的绿色荧光蛋白(adenovirusmediatedgreenfluorescentprotein,Ad-GFP,4组:重组腺病毒自身对照组)和DMEM培养液(5组:空白对照组),于注射后第7天手术,术后第7天检测各组大鼠TRAM皮瓣成活面积、CD34微血管密度(microvasculardensity,MVD)、VEGF免疫组织化学染色和原位杂交组织化学(insituhybridizationhistochemistry,ISHH)。结果11～3组TRAM皮瓣成活面积分别为14.19±2.77、15.18±2.18、8.30±1.28cm2,均较4、5组4.12±1.86、3.60±1.95cm2明显增加,且差异有统计学意义(P<0.05);21～3组TRAM皮瓣CD34MVD均高于4、5组,差异有统计学意义(P<0.05);31～3组VEGF免疫组织化学染色和ISHH呈阳性表达。结论重组腺病毒载体介导的VEGF165基因局部注射,能增加大鼠TRAM皮瓣局部缺血区域成活面积。

Objective To investigate the enhancement of the transverse rectus abdominis musculocutaneous （TRAM） flap survival in local ischemic area by recombinant adenovirus mediated vascular endothelial growth factor 165 gene （Ad-VEGF165）. Methods The vascular pedicle TRAM flaps were made in the right abdomin of 30 SD rats and they were randomly divided into 5 groups. The Ad-VEGF165 was injected into the subcutaneous tissue of epigastra（group 1 ）, the subcutaneous tissue of epigastria and rectus abdominis muscle （group 2）, and the rectus abdominis muscle（group 3）; Adenovirus mediated green fluorescent protein（Ad-GFP） and DMEM were injected into the subcutaneous tissue of epigastria and rectus abdominis muscle as autocontrol （group 4） and blank control （group 5）, respectively. The survival areas of TRAM flap was measured after 7 days of operation. The microvascular density（MVD） and the integral optical density （IOD） were tested with anti-rat CD34 and with VEGF immunohistochemistry and in situ hybridization histochemistry （ISHH）, respectively. Results The survival areas of TRAM flap in groups 1, 2 and 3 （14.19±2.77,15.18±2.18 and 8.30±1.28 cm^2） were higher than those in groups 4 and 5（4.12±1.86 and 3.60±1.95 cm^2）, being significant differences（P〈0. 05）. The CD34 MVD of the TRAM flap in groups 1, 2 and 3 was higher than that in groups 4 and 5 ; the positive expression for VEGF and ISHH were shown in groups 1, 2 and 3 and there was significant difference when compared with groups 4 and 5 （P〈0.05）. Conclusion Treatment by recombinant Ad-VEGF165 gene is an effective option for enhancement of the TRAM flap survival in the local ischemic area.