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目的在体研究切应力诱导内皮细胞TF基因表达变化规律及其机制。方法54只SD大鼠随机分为对照组和颈动脉狭窄组,狭窄组有又分为0.5、1、3、6、12 h、1、3、7 d 8个时相点,套扎法建立左颈总动脉狭窄模型,术后不同时相点用原位杂交和免疫组化法检测TF、Egr、Sp-1的mRNA和蛋白,用图像分析系统测定内膜平均灰度,进行统计学分析。结果正常对照组内皮细胞TF、Egr-1、Sp-1的mRNA和蛋白弱表达,狭窄30 m in后,内皮细胞胞浆组织因子基因mRNA转录和蛋白合成升高,与对照组比较均有显著性差异(P<0.05)。内皮细胞胞核和胞浆的Sp-1和Egr-1基因mRNA转录和蛋白合成均有显著性增加(P<0.05),但以Egr-1增加更显著(P<0.05),其变化趋势与组织因子基因mRNA转录、组织因子基因蛋白合成的变化趋势相同。TF mRNA和蛋白6 h达到峰值,Egr-1 mRNA和蛋白3 h达到峰值,Sp-1 mRNA和蛋白1 h达到峰值,与邻近组比较差异显著(P<0.05)。结论切应力是内皮细胞组织因子基因表达上调的触发因素之一,切应力激活TF与转录因子Egr-1和Sp-1介导有关。
Objective To study the changes of TF gene expression in endothelial cells induced by shear stress and its mechanism in vivo. Methods Fifty-four SD rats were randomly divided into control group and carotid artery stenosis group. The stenosis group was divided into 0.5, 1, 3, 6, 12 h, 1, 3, 7 d 8 time points, Left and right common carotid artery stenosis model, mRNA and protein of TF, Egr and Sp-1 were detected by in situ hybridization and immunohistochemistry at different time points after operation. Mean gray value of endometrium was measured by image analysis system and statistically analyzed . Results The mRNA and protein expressions of TF, Egr-1 and Sp-1 in endothelial cells were weakly expressed in normal control group. After 30 min of stenosis, mRNA transcription and protein synthesis of cytosolic factor in endothelial cells were increased, which were significantly higher than those in control group Sex differences (P <0.05). The mRNA and protein synthesis of Sp-1 and Egr-1 in the nucleus and cytoplasm of endothelial cells were significantly increased (P <0.05), but increased more significantly with Egr-1 (P <0.05) Tissue factor mRNA transcription, tissue factor gene protein synthesis trend of change. The mRNA and protein of TF reached the peak at 6 h, the mRNA and protein of Egr-1 peaked at 3 h, and peaked at 1 h of Sp-1 mRNA and protein, which was significantly different from the adjacent group (P <0.05). Conclusion Shear stress is one of the triggers of up-regulation of tissue factor gene expression in endothelial cells. Shear stress-activated TF is associated with the transcription factors Egr-1 and Sp-1.