钙离子超载对低温保存人肝细胞的损伤作用的研究

目的：研究分离培养的成人肝细胞在低温保存状态下（0-4℃）的钙离子代谢特点和钙离子超载的损伤作用机制。方法：胶原酶消化法制备分离的成人肝细胞，钙离子荧光探针Flou-3标记，流式细胞仪检测。结果：低温保存下，细胞内游离钙迅速升高，逐渐达到稳态水平，随细胞内钙离子的升高，细胞的存活率也逐渐下降，但在含0.5mmol/L CaCl2的保存液组，保存效果明显好于其它两组；在含1.5mmol/L CaCl2的保存液组，继发的钙离子超载峰值出现早，而且要高于原发的钙超载峰。结论：低温保存可以引起成人肝细胞的钙超载，细胞外钙是细胞钙超载的主要来源，继发超载是缺血再灌注损伤的主要原因。

Injurious effects of calcium overload on human hepatocytes during cold storage

Objective To explore mechanism of hypothermic injury by calcium overload and investigate calcium metabolism in human hepatocytes during cold storage. Methods Isolated human hepatocytes were labeled by flou-3, a fluorescent calcium indicator, and detected by flow cytometry. Results During cold storage, intracellular free calcium increased rapidly and reached a hemostasis slowly. As intracellular calcium increased, the cell survival rate decreased. But in 0.5 mmol/L CaCl 2 group, the effect of preservation was better than in other 2 groups. In 1.5 mmol/L CaCl 2 group, the peak of secondary calcium overload was much earlier and higher than that of primary one. Conclusion Hypothermic preservation might cause human hepatocyte calcium overload. The extracellular calcium is a chief source of calcium. The secondary calcium overload is the main cause of ischemic injury.