Adenovirus-mediated protein-kinase-GIα suppresses the hypoxia-induced proliferation and phenotype-switching of pulmonary arterial smooth muscle cell
10.3760/cma.j.issn.0578-1426.2010.05.006
- VernacularTitle:野生型蛋白激酶GIα腺病毒抑制低氧肺动脉平滑肌细胞表型转换及细胞增殖
- Author:
Bin YI
;
Junyu LU
;
Li BAI
;
Guansong WANG
;
Guisheng QIAN
- Publication Type:Journal Article
- Keywords:
Myocytes,smooth muscle;
Protein kinases;
Adenoviridae
- From:
Chinese Journal of Internal Medicine
2010;49(5):385-388
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the proliferation and phenotype-switching of pulmonary arterial smooth muscle cell (PASMC) induced by hypoxia and interfered by Ad-PKGIα. And to investigate the potential regulative role of PKGIα gene in the molecule mechanism of hypoxia pulmonary vessel remodeling (HPVR). Methods To establish the pure PASMC cultured by tissue-sticking methods. Semi-quantitative reverse transcription and polymerase chain reaction (RT-PCR) and Western blot were used to examine the PKGIα mRNA and protein expression after PASMC were transfected by Ad-PKG. The mRNA and protein expressive change of smooth muscle α actin(SM-α-actin) determined the degree of cell phenotype-switching. The changes of PASMC proliferation were determined by flow cytometry and ~3H-TdR incorporated way. Results Ad-PKGIα could transfect into PASMC and highly express. Hypoxia down-regulated the expression of SM-α-actin protein (44. 25±5.34 in normoxia, 32. 18±4. 19 in 12 h hypoxia condition, 21.90 ±2. 44 in 24 h hypoxia condition, P < 0. 05), that could be blocked by the transfeetion of Ad-PKGIα. Hypoxia could push PASMC mitosis and proliferating(~3H-TdR incorporated way: 7570 ± 371 in normoxia,12 020± 831 in 12 h hypoxia condition,14 924 ± 1491 in 24 h hypoxia condition, P <0. 05), that could be blocked by the transfection of Ad-PKGIα, too. Conclusions The results suggested that PKGIα signaling pathway might play an important role in the molecule mechanism of HPVR. And PKGIα gene might be a target point of gene therapy.
