目的:探讨 Ad-SPK1 是否促进胃黏膜上皮细胞增殖和迁移。 方法:分离培养胎兔胃黏膜上皮细胞, 通过重组腺病毒介导鞘氨醇激酶在上皮细胞中的表达,western blot 及 SPK 酶活性检测其表达情况。 结果:体外 实验中,Western blot 和SPK 酶活性检测外源SPK1 在上皮细胞的表达情况。 用抗FLAG 的抗体在Ad-SPK1 感染 的上皮细胞检测到一条特异条带。 SPK 酶活性法检测不同MOI 为100 的Ad-SPK1 感染兔上皮细胞后SPK 的活 性最高。 结论:腺病毒介导的鞘氨醇激酶可促进实验兔胃溃疡黏膜上皮修复和溃疡面血管增生,加快溃疡愈合。 关键词:胃黏膜;鞘氨醇激酶;上皮细胞

To determine the effect of SPK1 gene transfer on the healing of acute stress gastric ulcers in a rabbit model. Methods:Separate training tire rabbit gastric mucosal epithelium cells, The restructuring of adeno virus mediated scabbard ammonia alcohol kinase in the expression of epithelial cells Western blot and SPK enzyme activity detection its. Results:Freshly isolated gastric mucosal epithelium cells from pregnant rabbit. Western blot analysis of cytosolic proteins from SPK1- transfected rat mesothelial cells using anti-FLAG antibody revealed a specific protein band that was absent in Ad -GFP -infected cells. Assay the cellular SPK activity of infected rat mesothelial cells. Densities of signals weredetermined by densitometry and found MOI for 100. Conclusions:Denovirus- mediated SPK1 transfer could promote gastric mucosa repair and new vascular formation, thus accelerating the healing of ulcers in experimental rabbit.

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