Zhao Yan, Xie Yuan, Chen Xian, Xu Wenjie, Wang Yan, Zhou Jianjiang
Key Laboratory of Molecular Biology, Guiyang Medical College, Guiyang 550004, China.
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Zhonghua Bing Li Xue Za Zhi. 2014 Dec;43(12):820-6.
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.
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.
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.
H.pylori infection induces the expression of transgelin, ATP synthase and lactate dehydrogenase, implying possible roles in the pathogenesis of gastric diseases including cancer.
通过长期感染幽门螺杆菌（H.pylori）建立胃癌动物模型，并通过蛋白质组学分析阐明其发病机制。
选取50只雄性蒙古沙鼠（4 - 5周龄，体重60 - 100 g）感染幽门螺杆菌，在感染后3、6、12和24个月获取胃组织。通过胃组织切片的苏木精-伊红（H-E）染色评估组织学变化。通过新鲜胃组织样本的体外培养、幽门螺杆菌16s rRNA的聚合酶链反应（PCR）扩增及银染色定位检测幽门螺杆菌。此外，在不同感染时间点对从胃组织样本中提取的蛋白质进行二维电泳（2-DE）。选择在幽门螺杆菌感染过程中数量增加的蛋白质斑点，通过液相色谱-串联质谱（LC-MS/MS）进行分析。最后，通过实时逆转录-聚合酶链反应（RT-PCR）分析人胃癌组织样本和淋巴结之间差异表达的蛋白质。
早在幽门螺杆菌感染后3个月就在沙鼠胃组织中观察到幽门螺杆菌定植，并持续至24个月。对感染动物的病理检查显示出各种组织学变化，包括急性胃炎、萎缩性胃炎、肠化生和胃癌。通过蛋白质组学分析鉴定出78种差异表达的蛋白质，其中36种蛋白质上调，42种蛋白质下调。经LC-MS/MS分析，鉴定出10种蛋白质，包括乳酸脱氢酶、ATP合酶、脂肪酸结合蛋白、COX5B、过氧化物还原酶-4、过氧化物还原酶、转胶蛋白、琥珀酰辅酶A连接酶、角蛋白和蛋白质二硫键异构酶A2，其中转胶蛋白、ATP合酶和乳酸脱氢酶在人胃癌和淋巴结中高表达。
幽门螺杆菌感染诱导转胶蛋白、ATP合酶和乳酸脱氢酶的表达，这意味着它们在包括癌症在内的胃部疾病发病机制中可能发挥作用。
