Exogenous administration of heme oxygenase-1 by gene transfer provides protection against hyperoxia-induced lung injury. Academic Article uri icon

Overview

MeSH

  • Adenoviridae
  • Animals
  • Apoptosis
  • Bronchoalveolar Lavage Fluid
  • Gene Expression Regulation, Enzymologic
  • Gene Transfer Techniques
  • Heme Oxygenase-1
  • Immunohistochemistry
  • Oxidative Stress
  • Oxygen
  • Pleural Effusion
  • RNA, Messenger
  • Rats
  • Rats, Sprague-Dawley
  • Superoxide Dismutase

MeSH Major

  • Heme Oxygenase (Decyclizing)
  • Hyperoxia
  • Lung

abstract

  • Heme oxygenase-1 (HO-1) confers protection against a variety of oxidant-induced cell and tissue injury. In this study, we examined whether exogenous administration of HO-1 by gene transfer could also confer protection. We first demonstrated the feasibility of overexpressing HO-1 in the lung by gene transfer. A fragment of the rat HO-1 cDNA clone containing the entire coding region was cloned into plasmid pAC-CMVpLpA, and recombinant adenoviruses containing the rat HO-1 cDNA fragment Ad5-HO-1 were generated by homologous recombination. Intratracheal administration of Ad5-HO-1 resulted in a time-dependent increase in expression of HO-1 mRNA and protein in the rat lungs. Increased HO-1 protein expression was detected diffusely in the bronchiolar epithelium of rats receiving Ad5-HO-1, as assessed by immunohistochemical studies. We then examined whether ectopic expression of HO-1 could confer protection against hyperoxia-induced lung injury. Rats receiving Ad5-HO-1, but not AdV-betaGal, a recombinant adenovirus expressing Escherichia coli beta-galactosidase, before exposure to hyperoxia (>99% O2) exhibited marked reduction in lung injury, as assessed by volume of pleural effusion and histological analyses (significant reduction of edema, hemorrhage, and inflammation). In addition, rats receiving Ad5-HO-1 also exhibited increased survivability against hyperoxic stress when compared with rats receiving AdV-betaGal. Expression of the antioxidant enzymes manganese superoxide dismutase (Mn-SOD) and copper-zinc superoxide dismutase (CuZn-SOD) and of L-ferritin and H-ferritin was not affected by Ad5-HO-1 administration. Furthermore, rats treated with Ad5-HO-1 exhibited attenuation of hyperoxia-induced neutrophil inflammation and apoptosis. Taken together, these data suggest the feasibility of high-level HO-1 expression in the rat lung by gene delivery. To our knowledge, we have demonstrated for the first time that HO-1 can provide protection against hyperoxia-induced lung injury in vivo by modulation of neutrophil inflammation and lung apoptosis.

publication date

  • April 1999

has subject area

  • Adenoviridae
  • Animals
  • Apoptosis
  • Bronchoalveolar Lavage Fluid
  • Gene Expression Regulation, Enzymologic
  • Gene Transfer Techniques
  • Heme Oxygenase (Decyclizing)
  • Heme Oxygenase-1
  • Hyperoxia
  • Immunohistochemistry
  • Lung
  • Oxidative Stress
  • Oxygen
  • Pleural Effusion
  • RNA, Messenger
  • Rats
  • Rats, Sprague-Dawley
  • Superoxide Dismutase

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC408257

Digital Object Identifier (DOI)

  • 10.1172/JCI5342

PubMed ID

  • 10194478

Additional Document Info

start page

  • 1047

end page

  • 1054

volume

  • 103

number

  • 7