不同孵育时间的红细胞裂解物对大鼠基底动脉平滑肌细胞内游离钙的影响。

Effects of erythrocyte lysate of different incubation times on intracellular free calcium in rat basilar artery smooth-muscle cells.

作者信息

Guan Y Y, Weir B K, Marton L S, Macdonald R L, Zhang H

机构信息

Department of Surgery, Pritzker School of Medicine, University of Chicago, Illinois, USA.

出版信息

J Neurosurg. 1998 Dec;89(6):1007-14. doi: 10.3171/jns.1998.89.6.1007.

DOI:10.3171/jns.1998.89.6.1007

PMID:9833828

Abstract

OBJECT

The purpose of this study was to characterize substance(s) in the erythrocytes that increase intracellular free Ca++ concentration ([Ca++]i) in smooth-muscle cells and that therefore may be involved in the pathogenesis of vasospasm.

METHODS

Because vasospasm occurs days after subarachnoid hemorrhage (SAH), the authors studied the effects of aged human erythrocyte hemolysate and its low-molecular-weight (LMW) and high-molecular-weight (HMW) fractions on [Ca++]i in freshly isolated rat basilar artery smooth-muscle cells. Fresh hemolysate (Day 0) produced a biphasic response consisting of a transient peak and a sustained plateau increase in [Ca++]i, whereas hemolysate prepared from cells incubated for 3, 7, or 14 days induced only a transient response without a sustained phase. The effect of hemolysate declined with increasing incubation time. The HMW fraction and purified human oxyhemoglobin (OxyHb) did not evoke a response. The LMW fraction from Days 3, 7, or 14 produced no response at low concentrations (< 10%) and a transient response at high concentrations (> 20%), and the effect diminished with increasing incubation time. Unfractionated hemolysate or the LMW fraction of hemolysate incubated for 21 days produced no response. The combination of the 10% LMW fraction from Day 3 plus the 10% HMW fraction (Days 3. 7, 14, or 21) transiently increased [Ca++]i,. However, [Ca++]i was not changed by the 10% LMW fraction from Day 14 plus the 10% HMW fraction from Day 3 or 14. In the presence of OxyHb, [Ca++]i was increased by the 10% LMW fraction on Days 3 and 7, but not by the LMW fraction from Days 14 or 21.

CONCLUSIONS

The decline over time in the effect of hemolysate on [Ca++]i indicates either that the time that substances are released from erythrocytes is important in the generation of vasospasm or that this experimental system as used is not representative of conditions present after SAH. The data indicate that the ability to elevate [Ca++]i in smooth-muscle cells with hemolysate is provided by multiple substances, including OxyHb. These substances may interact during specific times after incubation of erythrocytes in vitro.

摘要

目的

本研究旨在鉴定红细胞中能增加平滑肌细胞内游离钙离子浓度（[Ca++]i）、因而可能参与血管痉挛发病机制的物质。

方法

由于蛛网膜下腔出血（SAH）几天后会发生血管痉挛，作者研究了衰老的人红细胞溶血产物及其低分子量（LMW）和高分子量（HMW）组分对新鲜分离的大鼠基底动脉平滑肌细胞[Ca++]i的影响。新鲜溶血产物（第0天）产生双相反应，包括[Ca++]i的短暂峰值和持续平台期升高，而由培养3、7或14天的细胞制备的溶血产物仅诱导短暂反应，无持续期。溶血产物的作用随培养时间延长而下降。高分子量组分和纯化的人氧合血红蛋白（OxyHb）未引起反应。第3、7或14天的低分子量组分在低浓度（<10%）时无反应，在高浓度（>20%）时产生短暂反应，且其作用随培养时间延长而减弱。未分级的溶血产物或培养21天的溶血产物低分子量组分无反应。第3天10%的低分子量组分与第3、7、14或21天10%的高分子量组分联合使用可短暂增加[Ca++]i。然而，第14天10%的低分子量组分与第3或14天10%的高分子量组分联合使用时，[Ca++]i未改变。在有OxyHb存在的情况下，第3天和第7天10%的低分子量组分可增加[Ca++]i，但第14天或21天的低分子量组分则无此作用。