Effect of p21 gene transfection mediated by replication deficient adenovirus on the pulmonary hypertensive rat model.
- Author:
Shao-jie CHEN
1
;
Yi-biao WANG
;
Ou CHEN
;
Xiao-bo ZHU
;
Yu MA
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Animals; Hypertension, Pulmonary; genetics; therapy; Male; Muscle, Smooth, Vascular; Oncogene Protein p21(ras); genetics; Rats; Rats, Wistar; Transfection
- From: Chinese Journal of Pediatrics 2008;46(2):139-142
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the feasibility of transferring p21 gene into lung tissue with recombinant adenovirus with full-length cDNA of p21 inserted in the Wistar rat model of pulmonary hypertension (PAH) induced by left-to-right shunt, study the expression of the desired gene in vivo, find if overexpression of desired gene can inhibit pulmonary hypertension.
METHODSWith full-length cDNA of p21 transfected HEK293 cells with clonfectin, and was packed, amplified in order to obtain the high-titer recombinant adenovirus (AdCMV-p21). The infection titer was determined by TCID50 method and was diluted into 1.67 x 10(8) pfu/L. Wistar rats were randomly allocated to control group (n = 10), model group (n = 15), test group (n = 10) and test control group (n = 10). In model group and test group left-to-right shunt pulmonary hypertension was developed by using cuff technique. AdCMV-p21 was transfected into test group and test control group using tracheal inhalation. The mPAP, mRVP and RVHI were measured and compared between every two groups. The left lung was immunohistochemically stained to observe the result of transfection. The right lung was HE stained to observe morphological changes in arteria pulmonalis and calculate WT%.
RESULTSThe mRVP, mPAP and WT% in model group and test group were significantly higher than those in control (P < 0.05), which suggested that the rat model of PAH was established successfully. Brown spots in the nucleus of VSMCs of pulmonary artery were seen in test group and test control group, which indicated that AdCMV-p21 was transfected successfully. The rate of transfected cells in test group was (42.8 +/- 11.6)%, which was equal to that of test control group (P > 0.05). In test group, the mPAP was (20.06 +/- 3.40) mm Hg (1 mm Hg = 0.133 kPa), mRVP was (22.53 +/- 2.53) mm Hg, WT% was (30.8 +/- 3.5)%, which were significantly lower than those in model group (P < 0.05), but higher than those in control group and test control group (P < 0.05).
CONCLUSIONThe recombinant adenovirus could successfully carry p21 and transfect the lung tissue of PAH rat model, and full expression of p21. p21 gene could inhibit the development of PAH.
