Shigeta Kouichirou, Taniguchi Nobuyuki, Omoto Kiyoka, Madoiwa Seiji, Sakata Yoichi, Mori Masaki, Hatake Kiyohiko, Itoh Kouichi
Department of Clinical Laboratory Medicine, Jichi Medical School, Tochigi-ken, Japan.
J Ultrasound Med. 2003 Apr;22(4):365-73. doi: 10.7863/jum.2003.22.4.365.
We investigated whether an ultrasonic echo contrast agent containing microbubbles (Levovist [SH U 508A]; Schering AG, Berlin, Germany) could in routine use activate platelets.
Levovist and its main component, galactose, were mixed with separate samples of whole blood (1.5-75 mg/mL) from 5 healthy volunteers to form a 1-mL suspension sample. After in vitro exposure to ultrasound emitted from a commercial ultrasonic scanner at a pulse frequency of 3.5 MHz with a mechanical index of 1.9 and an exposure duration of 5 minutes, 5 microL of the sample was incubated for 20 minutes with the fluorescein isothiocyanate-labeled CD61 antibody, which is a platelet-specific antigen, and the phycoerythrin-labeled CD62P (P-selectin) antibody, an activation-specific antigen, both on the platelet surface. After more than 30 minutes of fixing in 1% paraformaldehyde, flow cytometric analysis was performed.
The percentage of CD62P-expressing platelets increased according to the concentrations of Levovist and galactose, which showed almost equal effects. Ultrasound exposure did not enhance the effect except at the highest concentration of Levovist (75 mg/mL).
In vitro, a galactose-based echo contrast agent could not activate the platelets at its routine concentration.
我们研究了一种含微泡的超声造影剂（Levovist [SH U 508A]；德国柏林先灵公司）在常规使用中是否能激活血小板。
将Levovist及其主要成分半乳糖与5名健康志愿者的全血单独样本（1.5 - 75 mg/mL）混合，形成1 mL悬浮样本。在体外以3.5 MHz的脉冲频率、1.9的机械指数和5分钟的暴露时间，用商用超声扫描仪发射的超声进行暴露后，取5 μL样本与异硫氰酸荧光素标记的CD61抗体（一种血小板特异性抗原）以及藻红蛋白标记的CD62P（P - 选择素）抗体（一种血小板表面激活特异性抗原）一起孵育20分钟。在1%多聚甲醛中固定30多分钟后，进行流式细胞术分析。
表达CD62P的血小板百分比随Levovist和半乳糖浓度增加而升高，二者效果几乎相同。除了Levovist最高浓度（75 mg/mL）外，超声暴露并未增强该效应。
在体外，基于半乳糖的超声造影剂在常规浓度下不能激活血小板。
