尖吻蝮蛇毒PCA对人脐静脉血管内皮细胞的影响

Effects of protein-C activator from Agkistrodon Acutus venom on human umbilical vein endothelial cells

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/m L 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/m L的PCA溶液和0.16、0.32、0.63、1.25μg/m L的PCA与0.1μg/m L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/m L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/m L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HUVEC损伤。

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 treatmem 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 p,g/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 200 μL 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. Conclusion .. Low concentration of PCA can, to a certain extent, reduce the injury of HUVECs caused by LPS.