CAJ | 학술논문

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5 ml含1×109 cfu TP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基因真核表达质粒的减毒沙门菌.治疗后21 d,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) mm2比(56±6)、(43±4)mm2],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.
목적 탐토감독사문균개도적간세포생장인자(HGF)기인치료대대서실험성위궤양적효과.방법 구건HGF진핵표체재체pcKH,장pcKH화대조질립pcK전입Ty21a감독사문균중,구건휴대pcKH화pcK적양성공정균TPH화TP.장40지Wistar대서수궤분위정상대조조、모형대조조、TP치료조화TPH치료조,매조10지;TP치료조、TPH치료조화모형대조조채용빙을산유도방법제비대서을산성위궤양모형,조모후제5천분별급여5 ml함1×109 cfu TP화TPH적10％ NaHCO3급동체적10％ NaHCO3용액관위,1차/d,공3차.치료결속후제21천육안급경하관찰위궤양유합정황,계산궤양면적、측량위점막상피적파행장도;검측HGF재위궤양점막조직중적표체수평.결과 성공구건휴대HGF기인진핵표체질립적감독사문균.치료후21 d,병리결과표명TPH치료조대서위점막상피증생명현,증생점막선체명현확장,궤양이유육아조직충전,차가견봉부적모세혈관;모형대조조화TP치료조궤양유합불명현.TPH치료조궤양면적명현소우모형대조조화TP치료조[(11 ±3) mm2비(56±6)、(43±4)mm2],차이유통계학의의(P＜0.05).TPH치료조점막상피증생파행장도명현대우모형대조조화TP치료조[(879±8) mm비(821±10)、(817±14) mm],차이유통계학의의(P＜0.05),TPH치료조위궤양점막조직중HGF표체수평명현고우모형대조조화TP치료조[(0.88 ±0.18) μg/g단백비(0.51 ±0.08)、(0.34 ±0.15) μg/g단백],차이유통계학의의(P＜0.05).결론 휴대HGF기인적감독사문균가촉진대서위궤양점막상피증생,가쾌궤양유합.
Objective To explore the effect of hepatocyte growth factor (HGF) mediated by attenuated salmonella on gastric ulcer iu rats.Methods The eukaryotic expression vector of HGF (pcKH) was constructed,then pcKH or control plasmid pcK was electrotransfected into reconbinant 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 intragastric administration of TP (1 × 109 cfu, dissolved in 10％ sodium bicarbonate) , TPH (1 × 109 efu, 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) mm2] (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 v s (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 ±0.08), (0.34±0.15) μg/g protein] (P ＜0.05).Conclusion Recombinant attenuated salmonella stain carrying HGF gene can significantly accelerate the healing of gastric ulcers via stimulating proliferation of gastric epithelial cells and angiogenesis of gastric ulcer wound.