减毒沙门菌介导肝细胞生长因子基因治疗大鼠胃溃疡的实验研究  

Effect of hepatocyte growth factor mediated by attenuated salmonella on gastric ulcer in rats

作　　者：哈小琴[1] 杨迎桂[1] 杨淑娟[1] 张萍[1] 薛荣利[1] 张媛媛[1] 郭文家[1] 肖娜娜[1] 

机构地区：[1]兰州军区兰州总医院检验中心甘肃省干细胞与基因药物重点实验室,730050

出　　处：《中国医药》2015年第12期1778-1782,共5页China Medicine

基　　金：国家自然科学基金（81273568）;甘肃省杰出青年基金（2011GS3841）;兰州军区后勤科研项目（CLZ1TJ09）

摘　　要：目的探讨减毒沙门菌介导的肝细胞生长因子（HGF）基因治疗对大鼠实验性胃溃疡的效果。方法构建HGF真核表达载体pcKH，将pcKH和对照质粒pcK转入Ty21a减毒沙门菌中，构建携带pcKH和pcK的阳性工程菌TPH和TP。将40只Wistar大鼠随机分为正常对照组、模型对照组、TP治疗组和TPH治疗组，每组10只；TP治疗组、TPH治疗组和模型对照组采用冰乙酸诱导方法制备大鼠乙酸性胃溃疡模型，造模后第5天分别给予5ml含1×10^9cfuTP和TPH的10％NaHCO3及同体积10％NaHCO3溶液灌胃，1次／d，共3次。治疗结束后第21天肉眼及镜下观察胃溃疡愈合情况，计算溃疡面积、测量胃黏膜上皮的爬行长度；检测HGF在胃溃疡黏膜组织中的表达水平。结果成功构建携带HGF基凶真核表达质粒的减毒沙门菌。治疗后21d，病理结果表明TPH治疗组大鼠胃黏膜上皮增生明显，增生黏膜腺体明显扩张，溃疡已由肉芽组织充填，且可见丰富的毛细血管；模型对照组和TP治疗组溃疡愈合不明显。TPH治疗组溃疡面积明显小于模型对照组和TP治疗组[（11±3）mm^2比（56±6）、（43±4）mm^2]，差异有统计学意义（P〈0．05）。TPH治疗组黏膜上皮增生爬行长度明显大于模型对照组和TP治疗组[（879±8）mm比（821±10）、（817±14）mm]，差异有统计学意义（P〈0．05），TPH治疗组胃溃疡黏膜组织中HGF表达水平明显高于模型对照组和TP治疗组[（0．88±0．18）μg／g蛋白比（0．51±0．08）、（0．34±0．15）μg／g蛋白]，差异有统计学意义（P〈0．05）。结论携带HGF基因的减毒沙门菌可促进大鼠胃溃疡黏膜上皮增生，加快溃疡愈合。Objective To explore the effect of hepatoeyte growth factor （HGF） mediated by attenuated salmonella on gastric ulcer in rats. Methods The eukaryotic expression vector of HGF （pcKH） was constructed, then peKH or control plasmid pcK was electrotransfected into recombinant attenuated salmonella Ty21a to constructed stain carrying pcKH （named as TPH strain） or pcK （ named as TP strain）. Totally 40 Wistar rats were randomly divided into normal group, model group, TP group and TPH group （n = 10 in each group）. Acetic acid-induced gastric ulcer was established in model group, TP group and TPH group, and was respectively given intragastrie administration of TP （ 1 x 109 cfu, dissolved in 10% sodium bicarbonate ） , TPH （ 1 × 10^9 cfu, dissolved in 10% sodium bicarbonate） and 10% sodium bicarbonate on the 5th after injection, 1 time per 3 days, for 3 times. The ulcer healing was observed on gross or microscopic, the ulcer area and crawl length of gastric epithelium were measured; the HGF expression in ulcer tissue was assessed. Results The recombinant attenuated salmonella stain carrying HGF expression vector was successfully constructed. After treatment, the pathology results showed that the gastric epithelium were obviously proliferated, the glands in proliferative gastric epithelium were obviously expanded and the ulcer was filled by granulation tissue with abundant capillary in TPH group; the ulcer healing was not obvious in model group and TP group. The ulcer area in TPH group was significantly smaller than that in model group and TP group [ （ 11 ± 3 ）mm2 vs （ 56 ±6 ）, （ 43 ± 4 ） mm^2] （ P 〈 0. 05 ） ; the crawling length of gastric epithelium in TPH group was significantly longer than that in model group and TP group [ （ 879 ±8） mm vs （821 ± 10）, （817 ± 14） mm] （P 〈0. 05）. The expression of HGF proteins in gastric ulcer tissue was significantly higher in TPH group than that in model group and TP group [（0. 88 ± 0. 18） μg/g protein vs （0.51

关 键 词：肝细胞生长因子 基因治疗 减毒沙门菌 胃溃疡 

分 类 号：R714.2[医药卫生—妇产科学]