Adeno-p53转染对高肺血流肺动脉高压大鼠的影响

Effect of adenovirus-mediated p53 gene transfection on rats with pulmonary hypertension induced by high pulmonary flow

目的探讨腺病毒介导野生型p53基因转染对高肺血流肺动脉高压大鼠肺动脉压力及平滑肌细胞的影响。方法 60只大鼠随机分为空白对照组、模型对照组、实验组和实验对照组,每组15只。模型对照组、实验组和实验对照组采用套管法连接右侧颈总动脉和颈外静脉建立高肺血流肺动脉高压大鼠模型。空白对照组仅做假手术暴露颈总动脉处理。采用气管吸入法转染携带野生型p53基因的腺病毒(Adeno-p53)至实验组,同法转染重组腺病毒载体(Adeno-lacZ)至实验对照组,病毒保存液转染至空白对照组和模型对照组。采用Western blot法分析p53基因的表达;采用右心导管测压法测定各组大鼠右室平均压(mRVP)、肺动脉平均压(mPAP);肺组织HE染色观察肺小动脉形态学改变,计算管壁厚度与血管外径比值(WT%)、凋亡指数(AI)等,评价野生型p53基因转染对大鼠肺动脉压力及平滑肌细胞凋亡的影响。结果实验组大鼠成功转染Adeno-p53后,大鼠mRVP、mPAP、RVHI、WT%低于其他三组(P<0.05),肺动脉平滑肌细胞AI高于其他三组(P<0.05)。结论 Adeno-p53基因转染能在一定程度上缓解高肺血流型大鼠肺动脉高压的发展,其机制可能与野生型p53基因可以诱导肺动脉平滑肌细胞凋亡的增加有关。

Objective To investigate influences of transfection of adenovirus-mediated wild type p53 gene（Adeno-p53） on the pressure of the pulmonary artery and vascular smooth muscle cells in pulmonary hypertensive rats induced by high pulmonary flow.Methods Sixty rats were randomly divided into four groups： the blank contrast group,the model contrast group,the experimental group,and the experimental contrast group,fifteen rats in each group.The pipe-in-pipe method was used to build the rat model of pulmonary hypertension induced by high pulmonary flow in the model contrast group,the experimental group and the experimental contrast group.The blank contrast group was given a sham operation.Adeno-p53 was transferred into the experimental group using tracheal inhalation,Adeno-lacZ into the experimental contrast group,and virus conversation into the other two groups.Expression of the p53 gene was analyzed by Western blot.The mean right ventricle pressure（mRVP）,mean pulmonary arterial pressure（mPAP）,percentage of wall thickness/vascular diameter（WT%） of the arteriole,and apoptosis were measured.Influences of transferred Adeno-P53 on the pressure of the pulmonary artery and vascular smooth muscle cells in model rats were estimated.Results Adeno-p53 was successfully transferred into rats with pulmonary hypertension induced by high pulmonary flow.The mRVP,mPAP,RVHI and WT% in the experimental group were lower than those in the other three groups（P0.05）, while the apoptosis index（AI） was higher（P0.05）.Conclusion Transfection of the p53 gene can restrain the development of pulmonary hypertension,which may be related to apoptosis of vascular smooth muscle cells in the pulmonary artery induced by p53.