Effect of intramuscular injection of hepatocyte growth factor plasmid DNA with electroporation on bleomycin-induced lung fibrosis in rats.
BACKGROUND
So far, there is no efficient treatment for pulmonary fibrosis. The objective of this study was to determine whether intramuscular injection of the hepatocyte growth factor (HGF) plasmid DNA by in vivo electroporation could prevent bleomycin-induced pulmonary fibrosis in rats, and to investigate the possible mechanisms.


METHODS
Twenty male Wistar rats were randomly divided into four groups: control group (group C), model group (group M), early intervention group (group I) and late intervention group (group II). Groups M, I and II were intratracheally infused with bleomycin, then injected the plasmid pcDNA3.1-hHGF to group I on day 7, 14 and 21. Group II received the same treatment like Group I on day 14 and 21. All the rats were killed on day 28 after bleomycin injection. We detected Homo HGF expression in the rats with ELISA method and estimated the pathological fibrosis score of lung tissue using hematoxylin eosin (HE) and Massion staining. The mRNA expression of transforming growth factor-beta1 (TGF-beta1), cycloxygenase-2 (COX-2), and rat HGF in rat pulmonary parenchyma were evaluated by RT-PCR. Immunohistochemistry and Western blotting were performed to determine the protein expression of transforming TGF-beta1 and COX-2 in lung parenchyma.


RESULTS
The plasmid pcDNA3.1-hHGF could express hHGF in NIH3T3 cells and the hHGF protein is secreted into the culture medium. The expression of hHGF protein could be monitored in quadriceps muscle, plasma and lung in Groups I and II. Pulmonary fibrosis levels of Groups I and II were obviously lower than that of group M (P < 0.05). Expression of TGF-beta1 protein and mRNA in lung tissue was markedly decreased in Groups I and II compared with Group M (P < 0.05). The level of expression of HGF and COX-2 mRNA was higher in Groups I and II than in Group M (P < 0.05).


CONCLUSIONS
Injection of the plasmid pcDNA3.1-hHGF into skeletal muscle with electroporation has a potential role in the treatment of bleomycin-induced lung fibrosis. Exogenous HGF may inhibit the expression of TGF-beta1 and regulate the crosstalk between AECs and mesenchymal fibroblasts.
Long
Xiang
X
Institute of Pulmonary Disease, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China.
Xiong
Sheng-dao
SD
Xiong
Wei-ning
WN
Xu
Yong-jian
YJ
D000818:Animals; D001761:Bleomycin / Q000633:toxicity*; D051546:Cyclooxygenase 2 / Q000235:genetics; D018274:Electroporation; D015316:Genetic Therapy; D017228:Hepatocyte Growth Factor / Q000235:genetics*; D006909:Hydroxyproline / Q000032:analysis; D007273:Injections, Intramuscular; D008168:Lung / Q000737:chemistry; D008297:Male; D018482:Muscle, Skeletal / Q000378:metabolism; D010957:Plasmids; D011658:Pulmonary Fibrosis / Q000139:chemically induced / Q000628:therapy*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D053773:Transforming Growth Factor beta1 / Q000032:analysis / Q000235:genetics