失血性休克大鼠小肠上皮细胞线粒体ATPase8亚基表达的变化
Expression of ATPase8 subunit gene in mitochondria DNA of small intestine epithelial cells of rats with hemorrhagic shock
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摘要: 目的 探讨正常与休克大鼠小肠上皮细胞线粒体DNA(mitochondria DNA,mtDNA)ATPase8亚基及其表达的变化,为休克的防治提供理论基础。方法 用透射电镜观察大鼠小肠上皮细胞线粒体形态学变化,用基因重组、测序,通过gene Bank与mtDNA已知序列进行比较,研究大鼠小肠上皮细胞mtDNA ATPase8基因的改变。用RT-PCR观察大鼠小肠上皮细胞线粒体ATPase8 mRNA的表达。结果 休克组大鼠小肠上皮细胞线粒体数密度和面密度与对照组相比有显著差异(P<0.05或P<0.01)。正常组小肠上皮细胞mtDNA ATPase8亚基9个DNA测序有2处突变(A7868G,空7767-7768G)。休克组有3处突变,即A7797空,T7772C、T7863C。失血性休克组大鼠上皮细胞线粒体ATPase8 mRNA表达比正常组显著性下降(P<0.05),为对照组的27.3%。结论 休克大鼠小肠上皮细胞mtDNA ATPase8基因突变以及表达水平下降是导致ATPase8亚基改变的重要原因。Abstract: Objective To provide the theoretical basis for the prevention and treatment of hemorrhagic shock by studying the expression of ATPase8 subunit gene in mitochondria DNA（mtDNA） of small intestinal epithelial cells of rats with hemorrhagic shock. Methods Morphology of mtDNA of small intestinal epithelial cells of rats with hemorrhagic shock was observed under a transmission electronic microscope.Changes of ATPase8 gene in mtDNA of small intestinal epithelial cells of rats with hemorrhagic shock were studied by gene recombination and sequencing.Expression of ATPase8 mRNA in mtDNA of small intestinal epithelial cells of rats was detected by RT-PCR. Results The number density and surface density of mtDNA in small intestinal epithelial cells were significantly different between the shock and control groups（P＜0.05）.Mutation was observed in A7868G and non-7767-7768G of control group and in non-A7797,T7772C and T7863C of shock group.The expression level of mtDNA ATPase 8 in small intestinal epithelial cells was 27.3% lower in shock group than in control group（P＜0.01）. Conclusion Lower mutation and expression levels of mtDNA ATPase8 gene and mRNA in small intestinal epithelial cells of rats with hemorrhagic shock are the important reasons for the changes of ATPase8 subunit gene.
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