人血管内皮生长因子D表达载体促进大鼠肝脏血管形成的研究

Angiogenesis in rat liver after administration of expression vector for human vascular endothelial growth factor D

目的 人血管内皮生长因子D表达载体 (PCHO/hVEGF D)肠系膜上静脉注射后大鼠肝脏表达的维持时间以及血管形成效果观察。方法 健康级S D大鼠实验组 (2 4只 ) ,肠系膜上静脉注射PCHO/hVEGF D 15 0 μg/只 ,对照组 (18只 )同法注射等量生理盐水 ,然后行门静脉部分结扎术 ,制成门静脉高压症模型。术后 8、14、2 1d处死大鼠 ,取下肝脏标本 ,原位杂交法检测人血管内皮生长因子D(hVEGF D)mRNA的表达水平 ;Ⅷ因子法计算肝脏的血管计数。结果 实验 1、2、3组的hVEGF D的mRNA表达平均积分和阳性率分别为 6.83± 1.72 ,5 /6;3 .71± 2 .14 ,2 /7;2 .5 7± 2 .0 7,1/7;对照组极弱或无表达。第 1组与第 2组、第 3组的差异均有非常显著性 (t1/ 2 =2 .86,t1/ 3 =3 .99,P <0 .0 1)。第 2组和第 3组之间 ,差异无显著性 (t =1.0 16,P >0 .0 5 )。对照组未检测到表达。实验 1、2、3组的血管计数分别为 :12 .83± 4.0 2、8.2 0± 1.92 ,t =2 .3 5 ,P <0 .0 5 ;12 .71± 3 .14、8.80± 1.79,t =2 .49,P <0 .0 5 ;12 .0 0± 3 .74、7.67± 2 .5 0 ,t =2 .41,P <0 .0 5。 3组均显著高于相应对照组 ;实验组 3组之间血管计数差异无显著性。结论 裸基因肠系膜静脉注射PCHO/hVEGF D可以在门静脉部分结扎大鼠得到高效表达hVEGF

Objective To investigate the expression efficiency and duration and its angiogenesis in rat liver after intravenous injection of naked expression vector for human vascular endothelial growth factor D (PCHO/hVEGF-D) through superior mesenteric vein under the development of portal hypertension.Methods Forty-two healthy S-D rats were divided into two groups:experimental group ( n =24) receiving superior mesenteric intravenous injection of 150 μl of naked DNA and control group ( n =18) receiving the same volume of saline.The portal vein was partly ligated to make models of pre-hepatic portal hypertension.The samples of rat liver were collected 8,14,21 days after operation.The expression of hVEGF-D was detected at mRNA level and compared between the groups by means of in situ hybridization,and the mean number of capillaries was calculated microscopically by the method of Factor Ⅷ and compared statistically between two groups.Results The expression and positive rate of hVEGF-D mRNA in the experimental group were 6.83±1.72,5/6;3.71±2.14,2/7;2.57±2.07,1/7 respectively but none in control groups.(2)The average number of capillaries were statistically more in each study group than that of corresponding control groups (Group Ⅰ:12.83±4.02、8.20±1.92, t = 2.35, P < 0.05; Group Ⅱ:12.71±3.14、8.80±1.79, t =2.49, P <0.05;Group Ⅲ:12.00±3.74、7.67±2.50, t =2.41, P <0.05,Ⅱ,Ⅲ).There are no statistical differences between each study group.Conclusions Intravenous superior mesenteric injection of naked PCHO/hVEGF-D could be an efficient approach for VEGF gene transfer and with quite good effects of angiogenesis in rat liver.