重组腺病毒Ad-HGF基因治疗小鼠慢性阻塞性肺疾病的实验研究  

Gene therapy for COPD with hepatocyte growth factor mediated by recombinant adenovirus vector

作　　者：李允模[1] 弓削健太郎[2] 

机构地区：[1]江苏省苏北人民医院呼吸内科,江苏扬州225001 [2]日本岐阜大学医学部第二内科

出　　处：《中国现代医学杂志》2012年第5期31-34,共4页China Journal of Modern Medicine

摘　　要：目的探讨携带肝细胞生长因子基因的重组腺病毒Ad-HGF基因对小鼠慢性阻塞性肺疾病(COPD)模型的治疗作用。方法猪胰弹性蛋白酶(PPE)气管内滴注(4 u/只)复制小鼠COPD模型30只,随机分为模型对照组、Ad-HGF治疗组和Ad-Lac Z治疗组,每组10只。造模后3 d Ad-HGF治疗组气管内滴注Ad-HGF(3×1010particle/mouse);Ad-LacZ治疗组及模型对照组气管内分别滴注同剂量的Ad-Lac Z及同体积0.9%氯化钠溶液。每组5只实验动物分别于处理后7 d和14 d处死,光学显微镜下观察肺组织病理形态,测量肺泡平均截线长度(mean linear intercept,Lm);TUNEL法检测细胞凋亡;Ki-67免疫组织化学染色评价内皮细胞再生,并检测血浆HGF浓度。结果处理后7 d及14 d Ad-HGF治疗组血浆HGF浓度均明显高于Ad-LacZ治疗组及模型对照组,肺泡平均截线长度明显小于Ad-LacZ治疗组及模型对照组。TUNEL染色表明Ad-HGF治疗组与Ad-LacZ治疗组相比TUNEL阳性细胞明显减少。免疫组织化学染色显示Ad-HGF治疗组较Ad-LacZ治疗组Ki-67细胞明显增多。结论腺病毒介导的HGF基因可能有部分治疗小鼠COPD的作用。[Objective] To observe the effect of hepatocyte growth factor （HGF） on experimental chronic obstructive pulmonary disease （COPD） in mice,to investigate the mechanisms of the effect. [Methods] COPD model was made by porcine pancreatic elastase （PPE） in A/J mice, and a total of 30 mice were randomly divided into 3 groups： emphysema group, emphysema＋Ad-HGF group and emphysema＋ Ad-LacZ group, 10 mice each group. 3 days after induce emphysema modes emphysema＋ Ad-HGF group and emphysema＋Ad- LacZ group were intratracheally instilled with 3 xl0~~ particle Ad-HGF and Ad-LacZ respectively, and emphysema group mice same dose of saline were instilled. 7 days and 14 days after treatment and then killed the mice. The pathological sections of lung tissues from each group were prepared and observed under microscopy, the mean linear intercept （Lm） were calculated. Terminal deoxynucleotidy transferase-mediated dUTP nick end labeling （TUNEL） methods and Ki-67 immunohistochemical staining were carried out to examine the positive cells and distribution of cells regeneration. The plasma HGF concentration was measured. [Results] 7 days and 14 days after treatment, the results of plasma HGF concentration, Ki-67 positive cells showed significantly increased in emphysema＋ Ad-HGF group than emphysema＋ Ad-LacZ and emphysema group. Compared with emphysema＋Ad-LaeZ and emphysema group, the lung tissue of emphysema＋ Ad-HGF group had much less Lm and TUNEL positive cells. [Conclusion] The HGF may be a useful factor for the gene therapy on COPD in mice.

关 键 词：腺病毒 肝细胞生长因子 慢性阻塞性肺疾病 基因治疗 

分 类 号：Q784[生物学—分子生物学] R563[医药卫生—呼吸系统]