title: [Effects of sodium hydrogen exchanger-1 hammerhead ribozyme on proliferation of pulmonary artery smooth muscle cells in rats in vitro].
abstract: OBJECTIVE
  To 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.
  
  
  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 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.
  
  
  RESULTS
  Compared 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.
  
  
  CONCLUSIONS
  NHE-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.
authors.affiliation: Institute of Respiratory Disease of PLA, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Yao
authors.forename: Jun-Yu
authors.initials: JY
authors.lastname: Lu
authors.forename: Gui-Sheng
authors.initials: GS
authors.lastname: Qian
authors.forename: Xiao-Jing
authors.initials: XJ
authors.lastname: Yang
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D009131:Muscle, Smooth, Vascular; D032389:Myocytes, Smooth Muscle / Q000166:cytology*; D011651:Pulmonary Artery / Q000166:cytology* / Q000473:pathology; D016337:RNA, Catalytic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D000074082:Sodium-Hydrogen Exchanger 1; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*; D014162:Transfection
chemical_list: D016337:RNA, Catalytic; D012333:RNA, Messenger; C452850:Slc9a1 protein, rat; D000074082:Sodium-Hydrogen Exchanger 1; D017923:Sodium-Hydrogen Exchangers; C427204:hammerhead ribozyme