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目的 探讨内毒素 (LPS)对血管内皮细胞高能磷酸合成的影响及其分子基础。方法 通过抑制消减杂交克隆培养人脐静脉内皮细胞 (HUVEC)LPS刺激 6h后差异表达的基因 ,测序后得到一个下调表达的 2 0 3bp序列 ,与线粒体ATP合酶同源 ,扩增该序列并标记为探针 ,进行Northern杂交验证 ;测定胞内高能磷酸含量。结果 LPS刺激后 ,HUVEC内线粒体ATP合成酶基因表达下调 ,胞内ATP含量和能荷值显著降低。结论 LPS通过调控内皮细胞线粒体ATP合酶基因的表达 ,抑制线粒体呼吸功能 ,致细胞能量合成减少而影响细胞功能
Objective To investigate the effect of endotoxin (LPS) on the synthesis of high-energy phosphate in vascular endothelial cells and its molecular basis. Methods Genes differentially expressed 6 h after LPS stimulation of human umbilical vein endothelial cells (HUVECs) were cloned by suppression subtractive hybridization. A down-regulated 203 bp sequence was obtained after sequencing and homologous to mitochondrial ATP synthase. The sequence was amplified and labeled As a probe, Northern blot verification; Determination of intracellular high-energy phosphoric acid content. Results After stimulation with LPS, the expression of mitochondrial ATP synthase gene in HUVEC was down-regulated and the intracellular ATP content and energy charge decreased significantly. Conclusion LPS can regulate mitochondrial respiratory function by regulating the expression of mitochondrial ATP synthase gene in endothelial cells and decrease the cellular energy synthesis