Yazawa Koji, Isaka Yoshitaka, Takahara Shiro, Imai Enyu, Ichimaru Naotsugu, Shi Yi, Namba Yukiomi, Okuyama Akihiko
Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita 565-0871, Japan.
Nephrol Dial Transplant. 2004 Apr;19(4):812-6. doi: 10.1093/ndt/gfh064.
The clinical utility of cyclosporin A (CsA) has been limited by its nephrotoxicity, which is characterized by tubular atrophy, interstitial fibrosis and progressive renal impairment. Hepatocyte growth factor (HGF), which plays diverse roles in the regeneration of the kidney following acute renal failure, has been reported to protect against and salvage renal injury by acting as a renotropic and anti-fibrotic factor. Here, we investigated protective effects of HGF gene therapy on CsA-induced nephrotoxicity by using an electroporation-mediated gene transfer method.
CsA was orally administered as a daily dose of 30 mg/kg in male Sprague-Dawley rats receiving a low sodium diet (0.03% sodium). Plasmid vector encoding HGF (200 micro g) was transferred into the kidney by electroporation.
HGF gene transfer resulted in significant increases in plasma HGF levels. Morphological assessment revealed that HGF gene transfer reduced CsA-induced initial tubular injury and inhibited interstitial infiltration of ED-1-positive macrophages. In addition, northern blot analysis demonstrated that cortical mRNA levels of TGF-beta and type I collagen were suppressed in the HGF group. Finally, HGF gene transfer significantly reduced striped interstitial phenotypic alterations and fibrosis in CsA-treated rats, as assessed by alpha-smooth muscle actin expression and Masson's trichrome staining.
These results suggest that HGF may prevent CsA-induced tubulointerstitial fibrosis, indicating that HGF gene transfer may provide a potential strategy for preventing renal fibrosis.
环孢素A（CsA）的临床应用因肾毒性而受限，其特征为肾小管萎缩、间质纤维化和进行性肾功能损害。肝细胞生长因子（HGF）在急性肾衰竭后肾脏再生中发挥多种作用，据报道它作为一种促肾生长和抗纤维化因子可预防和挽救肾损伤。在此，我们采用电穿孔介导的基因转移方法研究了HGF基因治疗对CsA诱导的肾毒性的保护作用。
在接受低钠饮食（0.03%钠）的雄性Sprague-Dawley大鼠中，每天口服给予30mg/kg的CsA。通过电穿孔将编码HGF的质粒载体（200μg）导入肾脏。
HGF基因转移导致血浆HGF水平显著升高。形态学评估显示，HGF基因转移减轻了CsA诱导的初始肾小管损伤，并抑制了ED-1阳性巨噬细胞的间质浸润。此外，Northern印迹分析表明，HGF组中TGF-β和I型胶原的皮质mRNA水平受到抑制。最后，通过α-平滑肌肌动蛋白表达和Masson三色染色评估，HGF基因转移显著减少了CsA处理大鼠的条纹状间质表型改变和纤维化。
这些结果表明，HGF可能预防CsA诱导的肾小管间质纤维化，提示HGF基因转移可能为预防肾纤维化提供一种潜在策略。
