流动剪应力对丹酚酸B抑制血小板聚集的影响  

Influence of fluid shear stress on anti-platelet aggregation of salvianolic acid B

作　　者：常青[1,2] 郭宇飞 游云[1] 杨洪军[1] 梁日欣[1] 殷小杰[1] 李文[1] 廖福龙[1] 

机构地区：[1]中国中医科学院中药研究所,北京100700 [2]陕西中医学院药学院,西安712046

出　　处：《医用生物力学》2013年第3期344-349,共6页Journal of Medical Biomechanics

基　　金：国家自然科学基金资助项目(81274006)

摘　　要：目的研究流动剪应力对丹酚酸B抑制血小板聚集的影响及其可能的机制。方法采用生物力药理学的研究方法,2×4析因设计分组,利用BioFlux 1000控剪应力微流培养系统,分别施以0.02、1.5 Pa剪应力并联合4个剂量的丹酚酸B对血管内皮细胞(HUVECs)进行预处理20 h,收集上清液,采用联酶免疫吸附法(ELISA)检测细胞上清液中6-keto-PGF1α和vWF含量;检测上清液对ADP诱导的血小板聚集的影响;免疫荧光法分析流动内皮细胞胞浆vWF含量。结果与低剪应力流动条件相比,1.5 Pa剪应力联合100μg/mL丹酚酸B能显著提高内皮细胞分泌6-keto-PGF1α的量(P<0.05);细胞上清液明显抑制了血小板聚集(P<0.05)。剪应力的变化显著影响血管内皮细胞释放vWF(P<0.01),而丹酚酸B对血管内皮细胞释放vWF无明显影响。结论丹酚酸B可能通过促进血管内皮细胞分泌PGI2发挥抗血小板聚集的效应。从血流/血管/血液相互作用的角度看,正常的流动剪应力是丹酚酸B发挥抗血小板聚集效应的有利条件之一。Objective To study effects of flow shear stress combined with salvianolic acid B （ Sla B） on anti-plate- let aggregation and its possible mechanism under the theoretical framework of biomechanopharmacology. Meth- ods 2 x4 factor experimental design was employed. By using Bioflux 1000 microfluidic system, shear stresses of O. 02 Pa and I. 5 Pa were applied together with four levels of Sla B concentration treatment on human vascular endothelial cells （HUVECs） for 20 hours. Then the cell supernatant was collected to detect concentration of 6-ke- to-PGF1 α and vWF by ELISA and their effects on ADP-induced platelet aggregation were tested. Immunofluores- cence method was used to detect vWF in endothelial cell cytoplasm. Results Physical shear stress of l. 5 Pa combined with Sla B of 100 μg/mL could significantly promote the endothelial secretion of 6-keto-PGFlα as com- pared to low shear stress condition （ P 〈 O. 05 ）. The endothelial cell supernatant under shear stress of 1.5 Pa showed an obvious anti-platelet aggregation effect. As the single factor, shear stresses significantly influenced vWF secretion （ P 〈 0.01 ）, but Sla B had no obvious effects on vWF secretion. Conclusions Sla B inhibited ADP-induced platelet aggregation by increasing endothelial secretion of PGI2 under physical shear stresses. From the view of biomechaopharmacology （ interaction between blood flow, blood vessel and blood）, the physical flow shear stress is beneficial for the anti-thrombosis effect of Sla B.

关 键 词：剪应力 丹酚酸B 血管内皮细胞 血小板 

分 类 号：R318.01[医药卫生—生物医学工程]