Effect of portal hypertention by HGF gene therapy for liver cirrhosis

我们对肝硬变进行了基因治疗。每周连续3天给予二甲基亚硝胺(DIN)诱导大鼠和犬的肝硬变。DMN是一种肝毒性药物，会导致肝硬变。4周时肝细胞死亡，实质塌陷，纤维成分明显。在建立肝硬变模型后，将裸质粒HGF注射入肝动脉。裸质粒HGF不含任何病毒或脂类成分。HGF在基因转移后7~14天达到最大值。Azan-Mallory染色显示，注射HGF的大鼠肝纤维化区域明显减少，小叶结构排列整齐，而模型大鼠可见纤维间隔的形成和假小叶的形成。图像分析仪测量肝纤维化程度。肝细胞生长因子基因治疗的肝硬变肝纤维化程度较对照肝纤维化减轻50%以上。HGF治疗肝硬变的作用机制可能与其促进MMPs的产生和消化纤维组织有关。用免疫印迹法检测肝组织中基质金属蛋白酶-3的表达。在肝细胞生长因子基因转染后7d和14d，肝组织中这种蛋白的表达明显增加。这种治疗作用可抑制HGF处理的大鼠的门脉压力。肝细胞生长因子基因治疗改善了门静脉高压症大鼠的门脉高压，肝细胞生长因子治疗组大鼠的门脉压力低于模型对照组大鼠，裸鼠肝细胞生长因子基因转移到肝脏可以治疗大鼠肝硬变，最终可能转化为一种有效的治疗肝硬变的临床方案。

We performed gene therapy for liver cirrhosis. Liver cirrhosis was induced in rats and dogs by administrating dimethylnitrosamine (DIN) for 3 consecutive days a week. DMN is a hepatotoxic drug that causes liver cirrhosis. Hepatocyte death and parenchymal collapse occurred and fibrous component is evident by 4weeks. The naked HGF plasmid was injected into hepatic artery after cirrhosis has been established. Naked HGF plasmid does not contain any virus or lipid component. HGF showed maximum value from 7days to 14days after gene transfer. Azan-Mallory staining revealed that Regions of fibrosis markedly decreased and lobular architecture was well organized in HGF injected rats, but in MOCK rats, the formation of fibrous septa joining the central area and pseudolobule formation were evident. Photo-image analyzer measured the extent of fibrosis. Cirrhotic liver treated with HGF gene showed more than 50% less fibrosis than control cirrhotic liver. The proposed mechanism of therapeutic effect of HGF on liver cirrhosis is that HGF promotes MMPs producrtion and digests fiber tissue. We investigated expression of MMP-3 in the liver by Western blotting. The marked increase of this was observed in liver tissue of 7 days and 14 days after transfection of HGF gene. This therapeutic effect suppressed the portal pressure in HGF treated rats. HGF gene therapy improved the portal hypertension, Cirrhotic rat treated with HGF showed less portal pressure than MOCK transfected control cirrhotic rats.The naked HGF gene transfer into liver can treat rat liver cirrhosis and may be eventually translated into a useful clinical regimen for the treatment of patients with liver cirrhosis.