摘要
目的研究U937源性泡沫细胞形成过程中组织因子途径抑制物2的表达定位及变化规律。方法采用沉淀法提取人血浆低密度脂蛋白,硫酸铜氧化制备氧化型低密度脂蛋白,与U937单核细胞共同孵育,建立泡沫细胞模型,同时体外培养人脐静脉内皮细胞,加入U937泡沫细胞中共同培养,观察内皮细胞对泡沫细胞形成的影响。采用双重细胞免疫荧光定位组织因子途径抑制物2蛋白,时间免疫分辨荧光定量组织因子途径抑制物2蛋白,逆转录聚合酶链反应和荧光定量聚合酶链反应检测泡沫细胞内组织因子途径抑制物2基因的动态变化。结果组织因子途径抑制物2主要定位于U937单核细胞胞质中,与组织因子有相似的分布。氧化型低密度脂蛋白作用于U937细胞6h后组织因子途径抑制物2蛋白和基因表达水平持续增高,6h达至峰值而后表达持续降低。氧化型低密度脂蛋白作用于人脐静脉内皮细胞后组织因子途径抑制物2蛋白表达增加,24h时达至峰值。加入内皮细胞可改善氧化型低密度脂蛋白抑制U937细胞表达组织因子途径抑制物2的作用。结论U937泡沫细胞形成过程中,体外培养的人源性单核细株胞U937上存在组织因子途径抑制物2的表达抑制,而与人脐静脉内皮细胞共同培养可改善这种异常。
Aim To investigate the location and expression of tissue factor pathway inhibitor-2（TFPI-2） in U937 cell-derived foam cells.Methods Foam cell model was established by incubating the human monoblastic leukemia（U937） cells with oxidized low density lipoprotein（ox-LDL）.Double immuofluorescence was used to detect the location of TFPI-2 protein.Human umbilical vein endothelial cells（hUVEC） were incubated with U937,the effects of the expression of TFPI-2 on U937 cell-derived foam cells were observed.The expression of TFPI-2 protein was examined by time-resolved fluoroimmunoassay （TR-FIA）. The expression of TFPI-2 mRNA was examined by reverse transcription-polymerase chain reaction （RT-PCR） and realtime PCR. Results TFPI-2 protein distributed mainly in the cytoplasm of U937 cells. U937 cells were incubated with ox-LDL,the mRNA and protein of TFPI-2 peaked at 6 hours, then the levels of TFPI-2 decreased in a time dependent manner. After being added hUVEC, the expression of TFPI-2 in U937 increased. Conclusion Ox-LDL reduced the expression of TFPI- 2 in U937 cells, incubated with hUVEC can modify the inhibitive effect.
出处
《中国动脉硬化杂志》
CAS
CSCD
2007年第11期834-838,共5页
Chinese Journal of Arteriosclerosis
