Electroporation-mediated HGF gene transfer ameliorated cyclosporine nephrotoxicity.
BACKGROUND
The clinical utility of cyclosporine A (CsA) has been limited by its nephrotoxicity, which is characterized by tubular atrophy, interstitial fibrosis, and progressive renal impairment. Hepatocyte growth factor (HGF) has been reported to protect and salvage from renal injury as a renotropic and antifibrotic factor. Here, we investigated protective effects of HGF gene therapy on rat CsA-induced nephrotoxicity using electroporation-mediated gene transfer.


METHODS
CsA was subcutaneously administered daily under low sodium diet, and HGF gene was transferred into skeletal muscle by electroporation on days 7 and 14. We also examined the antiapoptotic mechanism of HGF using human proximal tubular epithelial cells.


RESULTS
HGF gene transfer rescued CsA-induced initial tubular injury and suppressed interstitial infiltration of ED-1-positive macrophages in CsA-induced nephrotoxicity. In addition, HGF significantly inhibited tubular cell apoptosis, and increased the number of proliferating tubular epithelial cells. In vitro studies suggest that HGF executes the antiapoptotic function by enhancing the phosphorylation of Akt and Bcl-2. Northern blot analysis demonstrated that HGF gene transfer suppressed cortical mRNA levels of transforming growth factor-beta (TGF-beta). Consequently, HGF gene transfer significantly reduced a striped interstitial phenotypic alteration and fibrosis.


CONCLUSIONS
We demonstrated that HGF gene transfer reduced CsA-induced tubular cell apoptosis and interstitial fibrosis. HGF gene transfer could be a potential strategy for preventing renal fibrosis.
Mizui
Masayuki
M
Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Isaka
Yoshitaka
Y
Takabatake
Yoshitsugu
Y
Mizuno
Sinya
S
Nakamura
Toshikazu
T
Ito
Takahito
T
Imai
Enyu
E
Hori
Masatsugu
M
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D016572:Cyclosporine / Q000633:toxicity*; D018274:Electroporation; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D017228:Hepatocyte Growth Factor / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000187:drug effects* / Q000293:injuries / Q000378:metabolism / Q000473:pathology; D007674:Kidney Diseases / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D008264:Macrophages / Q000187:drug effects / Q000473:pathology; D008297:Male; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley
D017228:Hepatocyte Growth Factor; D016572:Cyclosporine; C505407:CASP3 protein, human; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases