自制KYL液对大鼠肝脏低温保存后细胞凋亡影响的研究

目的 研究自制的KYL液对大鼠肝脏低温保存后细胞凋亡的影响。方法 采用大鼠肝脏非循环离体灌注模型（noncirculated isolated perfusion of ratliver，IPRL），随机以KYL液和UW液对大鼠肝脏保存0、4、8、16、24、48h，测定灌注流出液氧自由基代谢产物（丙二醛MDA和超氧化物歧化酶SOD）的含量，检测肝细胞内钙离子浓度，检测肝细胞凋亡率和凋亡相关基因表达，观察肝脏组织形态学变化。同时设生理盐水保存阴性对照组，了解器官保存液对大鼠肝脏有无保护作用。结果 KYL液保存的大鼠肝脏肝细胞内钙离子浓度较UW液保存者低，灌注流出液MDA和SOD含量与UW液保存者相近，两者肝细胞凋亡率及凋亡基因表达情况相近，光、电镜观察两者形态学变化基本一致。两组所有指标均较生理盐水保存组好，说明两种液体对大鼠肝脏均有保护作用。结论 自制的KYL液对大鼠肝脏的保存效果在钙拮抗方面略优于UW液，在抑制细胞凋亡方面与UW液相当，而在防止细胞水肿方面较UW液稍差。

Effects of cold preservation of rat liver with self-designed KYL solution on cell apoptosis

Objective To investigate the effects of cold preservation of rat liver with self-designed KYL solution on cell apoptosis. Methods Using the non-circulated isolated perfused rat liver (IPRL), SD rat liver were randomly preserved for 0, 4, 8, 16, 24 and 48 h with self-designed KYL solution or UW solution. The effects were assessed by measuring the content of free radical in Krebs-Henseleit's perfusate and content of intracellular calcium ion in the rat hepatocytes, observing the hepatocellular apoptosis and the related gene expression and the morphological changes in the liver. Meanwhile, the normal saline was used to establish the negative contrast model of rat liver preservation to determine whether the KYL solution and UW solution can prevent the ischemia-reperfusion injury in rat liver transplantation. Results The content of calcium ion in hepatocytes preserved by KYL solution was less than that in those preserved by UW solution. At different periods of preservation, the contents of MDA and SOD in Krebs-Henseleit's perfusate, the percentage of apoptotic cells and the expression of the apoptosis-related genes in liver tissue preserved by KYL solution were similar to those in that preserved by UW solution. Morphological changes were almost the same in both groups. All variables of the 2 groups were better than those in the rat liver tissue preserved by normal saline. Conclusions The self-designed KYL solution is similar to UW solution in effects on cell apoptosis.