重组腺病毒低氧诱导因子1α三突变体基因对兔缺血后肢血流灌注及血管渗透性的影响

Effects of Ad.HIF.1α.trip Gene on Blood Perfusion and Neovascular Permeability in Rabbit Hindlimb Ischemia

目的研究重组腺病毒低氧诱导因子1α三突变体基因(Ad-HIF-1α-trip)对兔缺血后肢血管渗透性的影响。方法 18只新西兰大白兔建立左侧后肢急性缺血模型,随机分成3组,分别为生理盐水组、腺病毒空载体组(Ad-Null组)、Ad-HIF-1α-trip组,每组6只,分别在术后即刻肌注生理盐水、Ad-Null及Ad-HIF-1α-trip。在术后即刻及基因转染后7、14和28 d进行血管超声检查。术后7 d、14 d和28 d通过血管超声计算缺血侧髂内动脉血流量标化值,评价兔缺血后肢血流灌注改善情况。基因转染后28 d采用伊文思蓝(EB)染色法测定各组缺血肌肉组织中EB含量,评价Ad-HIF-1α-trip对缺血组织新生血管渗透性的影响。结果 Ad-HIF-1α-trip组缺血侧髂内动脉血流量标化值增加大于其他2组,差异有统计学意义(P<0.05);Ad-HIF-1α-trip组缺血肌肉组织的EB含量略高于Ad-Null组及生理盐水组,但组间差异无统计学意义(P>0.05)。结论 Ad-HIF-1α-trip能有效改善缺血肌肉组织的血流灌注,同时并未明显增加新生血管的渗透性。

Objective To investigate the effects of the recombinant adenoviral expression of triple-point mutants HIFI-α gene （Ad-HIF- 1α-trip） on the blood perfnsion and neovascular permeability in the rabbit model of ischemic hincUimb. Methods After ligation of left femoral artery, 18 male New Zealand white rabbits were randomly divided into 3 groups ： saline group （ n = 6 ）, Ad - null group （ n = 6 ） and A d - HIF-1α- trip group （n = 6）, then the animals in different groups received saline, Ad-null and Ad-HIF-1α-trip respectively by intramuscular injection. Vascular ultra- sound was used to estimate the improvement of blood perfusion, the normalized blood flow of internal iliac artery of the hindlimb on the 7th, 14th, and 28th day after gene transfection was calculated. On the 28th day after gene tmnsfection, the permeability of the neovasculature in the iscbemic skeletal muscle was evaluated by Evans blue （EB） uptake assay. Results Vascular ultrasound revealed better blood perfnsion in A d-HIF- 1α- trip group than other groups （P 〈 0.05 ）. Quantification of the leakage levels of EB dye demonstrated that there were no obvious differences among three groups （P〉 0.05 ）, although the level of EB in the Ad-HIF-1α-trip group was a little higher than the other groups. Conclusion Ad-HIF-1α-trip can effectively improve the blood perfusion of ischemic limb, but did not significantly increase the permeability of the neovasculature.