摘要目的：构建幽门螺杆菌（ helicobacter pylori， HP）长期感染的动物模型，寻找与胃癌相关的差异表达蛋白，进一步阐明HP致胃癌的病理机制。方法蒙古沙鼠共50只，雄性，出生4～5周，体质量60～100 g，通过HP长期感染蒙古沙鼠构建动物模型，选择感染后3、6、12及24个月处死沙鼠，取胃组织，进行HP培养，同时用 PCR 扩增HP 16s rRNA基因及组织切片银染法检查HP感染定植情况，病理检查其胃组织病理改变。提取胃组织蛋白进行双向电泳，选取随时间点其蛋白表达呈增加趋势的差异蛋白质斑点进行液相色谱－质谱联用质谱鉴定，并检测差异蛋白基因在人胃癌及淋巴结组织中的表达。结果幽门螺杆菌感染3个月HP开始定植在鼠的胃组织，一直持续到24个月。病理检查可见急性胃炎、萎缩性胃炎、肠上皮化生及胃癌病理表现。蛋白组学分析共得到78个差异蛋白质，其中上调蛋白质36个，下调蛋白质42个，质谱鉴定出10个差异表达的蛋白质，包括乳酸脱氢酶、ATP合成酶、脂肪酸结合蛋白、细胞色素氧化酶C、过氧化物还原酶－4、过氧化物还原酶、transgelin、琥珀酰CoA连接酶、角蛋白－8、二硫化物异构酶A2。其中 transgelin、ATP 合成酶、乳酸脱氢酶基因在人胃癌和淋巴结组织高表达。结论 HP感染可能通过上调 transgelin、ATP合成酶、乳酸脱氢酶等基因的表达参与胃相关疾病的发生发展。

abstractsObjective To establish an animal model of gastric cancer by long-term infection of Helicobacter pylori ( H.pylori ) and to elucidate the pathogenesis by proteomics analysis.Methods Fifty male Mongolian gerbils (4-5 week-old and weighted 60-100 g) were infected with H.pylori and the gastric tissues were obtained after the infection at 3, 6, 12 and 24 months.Histological changes were evaluated by H-E staining of the gastric tissue sections.Detection of H.pylori was performed by in-vitro culture of fresh gastric tissue samples, PCR amplification of H.pylori 16s rRNA and localization by silver staining.In addition, proteins extracted from gastric tissue samples were subjected to two-dimensional electrophoresis (2-DE) at various infection time points.Protein spots with increased quantity over the course of H.pylori infection were selected and analyzed by LC-MS/MS. Finally, differentially expressed proteins between human gastric cancer tissue samples and lymph nodes were analyzed by real-time RT-PCR.Results Colonization of H.pylori was observed in gastric tissue of gerbils as early as 3 months after H.pylori infection, and persisted till 24 months. Pathological examination of infected animals showed various histological changes including acute gastritis, atrophic gastritis, intestinal metaplasia and gastric carcinoma. Seventy-eight differentially expressed proteins were identified by proteomics analysis, among which 36 proteins were up-regulated and 42 were down-regulated. Analyzed by LC-MS/MS, ten proteins were identified, including lactate dehydrogenase, ATP synthase, fatty acid-binding protein, COX5B, peroxiredoxin-4, peroxide reductase, transgelin, succinyl-CoA ligase, keratin and protein disulfide- isomerase A2, among which transgelin, ATP synthase and lactate dehydrogenase were highly expressed in human gastric carcinoma and lymph nodes.Conclusions H.pylori infection induces the expression of transgelin, ATP synthase and lactate dehydrogenase, implying possible roles in the pathogenesis of gastric diseases including cancer.