五步蛇毒蛋白C激活物对人脐静脉血管内皮细胞组织因子表达的影响  

Effects of Agkistrodon acutus venom protein C activator on expression of tissue factors in human umbilical vein endothelial cells

作　　者：靳文 张根葆[1,2] 高恬媛 俞玲 

机构地区：[1]皖南医学院病理生理学教研室,安徽芜湖241002 [2]皖南医学院蛇毒研究所,安徽芜湖241002

出　　处：《中国新药与临床杂志》2017年第11期684-688,共5页Chinese Journal of New Drugs and Clinical Remedies

基　　金：安徽省教育厅自然科学研究基金重点项目(KJ2011A266);活性生物大分子研究安微省重点实验室(科基[2012]126号)

摘　　要：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。AIM To observe the protective effects of Agkistrodon acutus venom protein C activator（ PCA） on human umbilical vein endothelial cells（ HUVEC）, and analyze PCA’ s effects for tissue factors（ TF） expression. METHODS HUVEC were conventionally cultured and divided into blank control group,lipopolysaccharide（LPS） group, PCA（0.625, 1.25, 2.50 μg·mL-1） group and PCA ＋ LPS group. MTT assay was used to detect the HUVEC viability. The secretion level of TF in HUVEC medium was performed by ELISA.Cellular localization of NF-κB was detected with immunofluorescence staining. RESULTS PCA had no significant effect on HUVEC activity and morphology（ P 〉 0.05）. The cell viability of LPS group was significantly decreased compared with the blank control group（ P 〈 0.01）, and the viability of LPS ＋ PCA group was significantly increased in the LPS ＋ PCA group than in the LPS group（P 〈 0.05）. The expression of TF and NF-κB were significantly increased in the LPS group than in the blank control group（P 〈 0.01）. But the expression of TF and NF-κB was significantly decreased in the LPS ＋ PCA group than in the LPS group（P 〈 0.01）. CONCLUSION PCA can protect HUVEC viability by reducing the secretion of TF in LPS-treated HUVEC, which may be associated with the suppression of NF-κB activation.

关 键 词：蛇毒液类 五步蛇毒蛋白C激活物 脂多糖类 NF-κB 凝血致活酶 

分 类 号：R996.3[医药卫生—毒理学]