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目的观察五步蛇毒蛋白C激活物(PCA)对人脐静脉血管内皮细胞(HUVEC)的保护作用,并分析其对组织因子(TF)表达的影响。方法常规培养HUVEC,实验分为空白对照组、脂多糖(LPS)组、PCA(0.625、1.25、2.50μg·mL~(-1))组和PCA+LPS组。MTT法检测HUVEC活性,ELISA检测HUVEC培养上清中TF分泌量,免疫荧光染色检测核转录因子(NF)-κB是否被激活-核转运。结果 PCA对HUVEC活性和形态无显著影响(P>0.05)。与空白对照组,LPS组细胞活性显著下降(P<0.01);而LPS+PCA组细胞活性显著高于LPS组(P<0.05)。LPS组TF表达量和NF-κB表达均显著高于空白对照组(P<0.01);LPS+PCA组与LPS组比较TF表达量和NF-κB表达降低(P<0.01)。结论 PCA可降低HUVEC分泌TF,减少LPS对HUVEC的损伤作用,其可能与抑制NF-κB通路的活化有关。
Objective To observe the protective effect of pentobarbital protein C activator (PCA) on human umbilical vein endothelial cells (HUVECs) and to analyze its effect on tissue factor (TF) expression. Methods HUVECs were cultured routinely. The experiment was divided into blank control group, lipopolysaccharide (LPS) group, PCA (0.625,1.25,2.50μg · mL -1) group and PCA + LPS group. The activity of HUVEC was detected by MTT assay. The secretion of TF in the supernatant of HUVEC was detected by ELISA. The nuclear translocation factor (NF) -κB was detected by immunofluorescence staining. Results PCA had no significant effect on the activity and morphology of HUVEC (P> 0.05). Compared with the control group, the cell viability in LPS group was significantly decreased (P <0.01), while the cell viability in LPS + PCA group was significantly higher than that in LPS group (P <0.05). The expression of TF and the expression of NF-κB in LPS group were significantly higher than those in blank control group (P <0.01). The levels of TF and NF-κB in LPS + PCA group were lower than those in LPS group (P <0.01). Conclusions PCA can reduce the secretion of TF and reduce the damage of HUVEC induced by LPS, which may be related to the inhibition of the activation of NF-κB pathway.