Xiong Bing, Yi Chuan-xun, Guo Jian-li, Zhang Yi-ming, Feng Xiao-ling, Zhou Wen-dang
Department of Burn Surgery, Guangdong People's Hospital, Guangzhou 510080, China.
Zhonghua Zheng Xing Wai Ke Za Zhi. 2003 May;19(3):211-3.
To investigate flap survival after transfection using gene encoding vascular endothelial growth factor (VEGF).
In 30 Sprague-Dawley rats, the anterior abdominal skin flap supplied by the epigastric vessels was created. The animals were divided into three groups, with ten of each. The first group was treated with a mixture of liposomes and the cDNA encoding the 165-amino acid isoform of VEGF; the second group was treated with control blank plasmid DNA and liposome transfection medium; the third group was treated with physiological saline. Four days after injection, the epigastric artery and vein were ligated and the blood flow in the flap was evaluated by intraperitoneal injection of fluorescence solution. Seven days later, the survival area of the flap was measured by planimetry. After the animals were killed, specimens were harvested from the anterior abdomen skin flap for immunohistological evidence of VEGF expression and for hematoxylin and eosin staining of microvascular growth.
30 minutes after pedicle ligation the average fluorescence staining planimetry of the three groups (PCD-VEGF165, PCD and physiological saline) was 60.64%, 30.15% and 29.89% respectively. Tissue survival planimetry of the three groups was 92.3%, 30.5%, 31.8%. There was significant difference between the first group and the latter two (P < 0.05). Immunohistochemical staining documented increased deposition of VEGF cDNA in the first group compared to the control groups (P < 0.05). Normal staining documented that the average vessel number of the three groups was 101.72, 91.35 and 89.85 (P < 0.05), the average vessel lumen diameter was 26 microns, 31.09 microns and 32.51 microns(P < 0.05).
Through administration, PCD-VEGF165 can transfect the anterior abdominal skin flap and enhance its survival. There was express of VEGF protein in the treated flap.
研究使用编码血管内皮生长因子(VEGF)的基因转染后皮瓣的存活情况。
在30只Sprague-Dawley大鼠中,制作由腹壁血管供血的腹部前皮瓣。将动物分为三组,每组10只。第一组用脂质体与编码VEGF 165个氨基酸异构体的cDNA混合物处理;第二组用对照空白质粒DNA和脂质体转染介质处理;第三组用生理盐水处理。注射4天后,结扎腹壁动静脉,并通过腹腔注射荧光溶液评估皮瓣中的血流。7天后,通过平面测量法测量皮瓣的存活面积。处死动物后,从前腹部皮瓣采集标本,用于VEGF表达的免疫组织学证据以及微血管生长的苏木精和伊红染色。
蒂部结扎30分钟后,三组(PCD-VEGF165、PCD和生理盐水组)的平均荧光染色平面测量值分别为60.64%、30.15%和29.89%。三组的组织存活平面测量值分别为92.3%、30.5%、31.8%。第一组与后两组之间存在显著差异(P<0.05)。免疫组织化学染色显示,与对照组相比,第一组中VEGF cDNA的沉积增加(P<0.05)。正常染色显示,三组的平均血管数量分别为101.72、91.35和89.85(P<0.05),平均血管腔直径分别为26微米、31.09微米和32.51微米(P<0.05)。
通过给药,PCD-VEGF165可转染腹部前皮瓣并提高其存活率。处理后的皮瓣中有VEGF蛋白表达。