重组人组织激肽释放酶结合蛋白抗氧化活性及抗凝活性分析  

Antioxidant Activity and Anticoagulant Activity Analysis of Recombinant Human Tissue Kallistatin Binding Protein

作　　者：周玥莹 龙莎 方美娟[2] 王立强 ZHOU Yueying;LONG Sha;FANG Meijuan;WANG Liqiang(School of Biomedical Sciences,Huaqiao University,Quanzhou 362021,China;School of Pharmaceutical Sciences,Xiamen University,Xiamen 361102,China)

机构地区：[1]华侨大学生物医学学院,福建泉州362021 [2]厦门大学药学院,福建厦门361102

出　　处：《华侨大学学报（自然科学版）》2018年第5期732-737,共6页Journal of Huaqiao University(Natural Science)

基　　金：国家重点研发计划资助项目(2016YFE0101700);国家自然科学基金资助项目(2015J01342)

摘　　要：探究Kallistatin蛋白在体外3种细胞模型中的抗氧化活性及其体外抗凝、溶栓活性.首先,在体外构建HBZY-1,HaCaT,HTR8-s/Vneo细胞的H2O2,Fe-HQ氧化应激模型,并将Kallistatin蛋白应用于构建的氧化应激模型中.然后,采用多浓度梯度法筛选最佳氧化剂浓度,利用细胞活性实验检测以上3种细胞的活性,并进行体外抗凝、溶栓实验考察.结果表明:在H2O2模型中,当Kallistatin蛋白浓度为0.6μmol·L-1时,3种细胞活力与对照组(LD-Hanks缓冲液)相近,低剂量蛋白组(0.06μmol·L-1)的细胞活力明显低于高剂量蛋白组细胞;在Fe-HQ模型中,不同细胞中Kallistatin蛋白剂量的作用不同,Kallistatin蛋白具有体外抗氧化活性,不同应激条件下不同细胞对蛋白剂量的依赖性不同;体外抗凝、溶栓实验表明此蛋白具有相关活性.To investigate the anti oxidative activity and anticoagulant and thrombolytic activity of Kallistatin in three kinds of cell models in vitro,H 2O 2 and Fe HQ oxidative stress models of HBZY 1,HaCaT and HTR8 s/Vneo cells were constructed in vitro,and the Kallistatin protein was used in these oxidative stress models.Multi concentration gradient method was adopt to select the optimal oxidant concentration.CCK experiment was used to detect the activity of three kinds of cells,and the anticoagulant and thrombolytic experiments were carried out in vitro.The results shows that in the H 2O 2 model,the viability of three kinds of cell was similar to the control group(LD hanks buffer)when the concentration of Kallistatin protein was 0.6μmol·L-1.The cell viability of low dose protein group(0.06μmol·L-1)was significantly lower than that of the high dose.In the Fe HQ model,the effect of Kallistatin protein doses differed in different cells.Thus,Kallistatin protein has antioxidant activity in vitro,and it is dose dependent in different cells under different stress conditions.Furthermore,in vitro anticoagulation and thrombolysis experiments showed that this protein has related activity.

关 键 词：Kallistatin蛋白 氧化应激 细胞活性 抗凝 溶栓 

分 类 号：R944[医药卫生—药剂学]