人肝细胞生长因子基因转染对大鼠肾脏缺血再灌注损伤的保护作用

Effects of hHGF gene electroporation on kidney ischemia-reperfusion injury in rat

目的用基因电转染方法分别在肾脏和骨骼肌进行人肝细胞生长因子（hHGF）基因转染,比较两种治疗模式对肾脏缺血再灌注损伤的保护作用。方法雄性SD大鼠30只,随机分为3组,每组10只,分别为对照组、肌肉电转染组及肾脏电转染组。在热缺血前3 d将含有hHGF基因的质粒用电转染的方法转入大鼠的骨骼肌和肾脏,3d后建立大鼠肾热缺血模型。观察hHGF的药代动力学,并评价缺血肾脏功能和组织学。结果肾脏电转染组的肾组织及血浆hHGF表达显著高于肌肉电转染组（P＜0．01）。在肾缺血后的第1、3、5天,两个电转染组的Scr水平均较对照组[（164．8±55．5）μmol/L]显著降低（P＜0．01）;而肾脏电转染组[（79．4±23．4）μmol/L]的Scr水平比肌肉电转染组[（109．7±18．6）μmol/L]显著降低（P＜0．05）。两个电转染组肾小管细胞坏死评分显著低于对照组（肾1．365±0．186、肌肉1．864±0．389、对照2．230±0．250,P＜0．05）;而肾脏电转染组的肾小管坏死评分显著低于肌肉电转染组（P＜0．05）。两个电转染组的淋巴细胞、巨噬细胞及中性粒细胞浸润均较对照组显著减少（P＜0．01）,而在两个电转染组间差异无统计学意义。结论无论是hHGF的药代动力学还是治疗效果,肾脏hHGF基因电转染均优于肌肉电转染。肾脏hHGF基因电转染有可能成为防治移植肾缺血再灌注损伤的有效方法之一。

Objective To compare the efficacy of two hHGF gene electroporation methodologies （muscle and kidney） in order to reduce the deleterious effects induced by renal warm ischemia. Methods Thirty male SD rats were divided into three groups, 10 in each group. One group was warm ischemia without pretreatment as control. The other two groups received hHGF gene muscle injection and subsequent electroporation and hHGF gene kidney electroporation respectively 3 days before renal warm ischemia. Pharmacokinetic, function and histology were assessed in ischemic kidney. Scr levels were measured at day 1,3,5 after injury. Results Plasma hHGF levels and hHGF expression of renal tissue in kidney electroporation group were higher than those in muscle electroporation group （P〈0.01）. Both treated groups showed lower Scr than control [（79.4±23.4）, （109.7±18.6） vs （164.8±55.5） μmol/L,P〈0.05]. The kidney electroporation group showed faster recovery of renal function, with significantly lower Scr level compared to muscle electroporation group. The tubular necrosis score was lower in both HGF-treated groups （P〈0.05）. The tubular necrosis score of the hHGF gene kidney electroporation group （1.365±0.186） was lower than that of hHGF gene muscle electroporation group （1.864±0.389） （P〈0.05）. hHGF-treated groups had fewer macrophages and lymphocytes than control group, as well as lower values of MPO activity（P〈0.01）, but no significant difference was found between 2 hHGF-treated groups（P〉0.05）. Conclusion Kidney direct electrotransfer is shown to be more efficient not only in pharmacokinetic but also in therapy as compared to muscle electrotransfer, so it may become a clinically practical alternative in renal transplantation.