摘要
目的：观察皖南尖吻蝮蛇毒蛋白C激活剂（ PCA）对人脐静脉血管内皮细胞（ HUVEC）的影响并分析其可能机制。方法：常规培养HUVEC，在倒置显微镜下观察细胞形态变化，用MTT法检测细胞活性。实验分为对照组、LPS损伤组和PCA实验组。将HUVEC接种在96孔板内加药处理，对照组加入PBS缓冲液，LPS损伤组加入0．1μg／mL LPS，PCA实验组分别加入0．16、0．32、0．63、1．25、2．50、5．00、10．00、20．00、40．00μg／mL的PCA溶液和0．16、0．32、0．63、1．25μg／mL的PCA与0．1μg／mL的LPS的混合液。每个浓度设5个复孔，每孔加药200μL，作用12 h后进行细胞活性测定并观察细胞形态。结果：PCA浓度低于2．50μg／mL时对HUVEC活性和形态没有明显影响，但可减轻LPS引起的HUVEC活性抑制和形态改变。高于2．50μg／mL时PCA可抑制HUVEC的活性并引起其形态改变。结论：低浓度PCA在一定程度上可减轻LPS引起的HU－VEC损伤。
Abstract
Objective:To observe the effects protein-C activator( PCA) extracted from the venom of Agkistrodon acutus on the human umbilical vein endo-thelial cells (HUVECs),and investigate the potential mechanisms of this effects.Methods:HUVECs were conventionally cultured,and observed under the inverted microscope for their morphologically changes .MTT method was used to detect the cell activity .The experiment consisted of control group ,LPS-in-duced injury group and PCA treatment group.HUVECs were initially inoculated into separate 96-well plate.Then control group were treated with PBS buff-er,LPS injury was induced with LPS in dose of 0.1 μg/mL,and PCA group were treated with PCA in dose of 0.16,0.32,0.63,1.25,2.50,5.00, 10.00,20.00 and 40.00 μg/mL and mixed solution of 0.1 μg/mL LPS and PCA dosed by 0.16,0.32,0.63 and 1.25 μg/mL,respectively.Five du-plicated wells were set for individual plate into which 200μL of the cell solution was added.After 12 hours of culturing,the cell activity and its morphology were determined and observed.Results:PCA level lower than 2.50 μg/mL had no effect on the activity of HUVECs,yet reduced the damage of LPS to HUVEC.Contrarily,PCA concentration over 2.50 μg/mL significantly inhibited the activity of HUVECs and modified their morphology .Conclusio n:Low concentration of PCA can,to a certain extent,reduce the injury of HUVECs caused by LPS.
基金项目
安徽省教育厅自然科学研究基金重点项目( KJ2011A266)
NETL
NSTL
维普
万方数据