Effects of hammerhead ribozyme on NHE-1 activity and proliferation in pulmonary artery smooth muscle cells of rats in vitro
- VernacularTitle:核酶对大鼠肺动脉平滑肌NHE-1活性及细胞增殖的作用
- Author:
Wei YAO
;
Junyu LU
;
Guisheng QIAN
;
Xiaojing YANG
;
- Publication Type:Journal Article
- Keywords:
hammerhead ribozyme;
sodium hydrogen exchanger;
pulmonary artery smooth muscle cell;
proliferation
- From:Journal of Third Military Medical University
1988;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of hammerhead ribozyme on the expression and activity of NHE 1 and pHi in pulmonary artery smooth muscle cells (PASMCs) of rats and its role in PASMCs proliferation in vitro . Methods According 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 hammerhead ribozyme, PRZ, was transfected into the cultured PASMCs. G418 resistant cell clones were screened with 100 ?g/ml G418. Then, the expression of NHE 1 mRNA was detected by RT PCR, intracellular pH(pHi) value and recovery rate of pHi after intracellular acid loading were measured by fluorescent probe BCECF. 22 Na uptake and 3H TdR incorporation were determined, respectively. Results Compared with the cells transfected with pLXSN and non transfected cells, NHE 1 mRNA, pHi value, pHi recovery rate, 22 Na uptake and 3H TdR incorporation decreased significantly in cells transfected with recombinant vector PRZ. No significance was found between the pLXSN transfected group and non transfected group. Conclusion Hammerhead ribozyme can cleave the target NHE 1 mRNA specifically, reduce the expression of NHE 1 mRNA, induce intracellular acidosis and consequently suppress the proliferation of PASMCs.
