幽门螺杆菌jhp947基因对C57BL/6小鼠致病性研究

Pathogenicity of Hp jhp947 gene to C57BL/6 mice?

摘要:

摘要：【目的】 检测jhp947对幽门螺杆菌(Helicobacter pylori,Hp )所致动物胃上皮细胞病变和基因表达的影响。【方法】 将27只无特定病原体的（specified-pathogens free，SPF）6 w 龄 C57BL/6小鼠随机分成3组，分别以J99 、△J99-947（缺失jhp947基因）和PBS灌胃，菌量为109CFU/mL，0、2、4 d各一次，4w后宰杀所有小鼠，无菌操作取胃组织，分别作快速尿素酶试验，Hp培养，组织病理学检查；免疫组化检测ESE-3A、NDRG1、MATP基因表达；半定量RT-PCR检测jhp947基因对小鼠ESE-3A、NADG1、MATP等基因转录的影响。【结果】 J99组和△J99-947组快速尿素酶试验和Hp培养的阳性率均为100%，PBS对照组快速尿素酶试验和Hp培养阴性。 组织病理学检查，PBS对照组小鼠胃黏膜100%正常， J99组小鼠胃黏膜33.3%（3/9）轻度糜烂，66.7%（6/9）重度糜烂。△J99-947组小鼠胃黏膜22.2%（2/9）正常，77.8%（7/9）轻度糜烂，没有重度糜烂。J99组小鼠胃黏膜的损伤比△J99-947组重。 免疫组化检测结果， J99组比△J99-947组ESE-3A、NDRG1、MATP表达显著降低（p<0.05）。 半定量RT-PCR检测结果， J99组比△J99-947组ESE-3A、NDRG1、MATP表达显著降低（p<0.05）。【结论】 JHP947基因存在可使Hp感染所致的胃黏膜损伤程度加重。在体内jhp947基因可能通过下调NDRG1、MATP等肿瘤抑制基因的表达，这可能与胃癌的发生有关。

Abstract:

Abstract [Objective] To study the effects of Helicobacter pylori jhp947 gene on the pathogenesis of epithelial cells and gene express pattern by animal studies . [Methods] Twenty-seven special pathogen free (SPF) C57BL/6 mice were divided equally into 3 groups, and challenged with Helicobacter pylori J99, Helicobacter pylori△J99-947 and phosphate buffer (PBS) respectively, at a dose of 109 colony formine unit (CFU) at 0, 2, and 4 days. Mice were sacrificed 4 weeks after the last challenge, and went through rapid urease test, culture of Hp, histological examination ,immunofluorescent histochemistry and semiquantitative reverse transcription PCR (RT-PCR) of gastric mucosa. [Results] The result of rapid urease test and culture of Hp indicated that the positive rates in J99 and △J99-947 group were both 100% while 0% in PBS group. The result of histology examination indicated that garstirc mucosa is all normal in PBS group; in J99 group, 33.3% (3/9） had slightly anabrosis, 66.7% (6/9) had seriously anabrosis; in △J99-947 group, 22.2% (2/9) is normal, 77.8% (7/9) had slightly anabrosis. The degree of anabrosis seems to be more severe in J99 than in △J99-947. The result of immunofluorescent histochemistry and semiquantitative RT-PCR of gastric mucosa indicated that the expression level of ets homologous factor, N-myc downstream regulated gene 1, methylthioadenosine phosphorylase is significantly lower in J99 than in △J99-947 group (P<0.05). [Conclusion] The degree of anabrosis seems to be more severe in Hp with jhp947 gene than in Hp without jhp947 gene. In vivo, jhp947 may induce tumorigenesis by inhibiting anti-oncogenes（N-myc downstream regulated gene 1 and methylthioadenosine phosphorylase）.