Effects of sodium hydrogen exchanger-1 hammerhead ribozyme on proliferation of pulmonary artery smooth muscle cells in rats in vitro.
- Author:
Wei YAO
1
;
Jun-Yu LU
;
Gui-Sheng QIAN
;
Xiao-Jing YANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Proliferation; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; cytology; Pulmonary Artery; cytology; pathology; RNA, Catalytic; genetics; RNA, Messenger; genetics; Rats; Sodium-Hydrogen Exchanger 1; Sodium-Hydrogen Exchangers; metabolism; Transfection
- From: Chinese Journal of Applied Physiology 2003;19(1):39-42
- CountryChina
- Language:Chinese
-
Abstract:
AIMTo explore the effects of sodium hydrogen exchanger (NHE-1) specific hammerhead ribozyme on the expression and activity of NHE-1 and pHi in pulmonary artery smooth muscle cells (PASMCs) in rats, and its role in PASMCs proliferation.
METHODSAccording to the secondary structure of NHE-1 mRNA in rats, NHE-1 specific hammerhead ribozyme was designed with the assistance of computer. The recombinant vector of retroviral plasmid pLXSN and NHE-1 hammerhead ribozyme, PRZ, was transfected into the cultured PASMCs. G418 resistant cell clones were screened with 60 microg/ml G418. Then, the expression of NHE-1 mRNA was detected by RT-PCR, intracellular pH was measured with fluorescent probe-BCECF, 22Na and 3H-TdR incorporation were determined respectively.
RESULTSCompared with the cells transfected with pLXSN and non-transfected cells, NHE-1 mRNA, pHi value, 3H-TdR and 22Na incorporation decreased significantly in cells transfected with recombinant vector PRZ. No significance was found between the pLXSN transfected group and non-transfected group.
CONCLUSIONNHE-1 hammerhead ribozyme can cleave the target RNA specifically, reduce the expression of NHE-1 mRNA, induce intracellular acidosis and consequently prohibit the proliferation of PASMCs.
