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定制的KYL溶液冷藏保存大鼠肝脏中的细胞凋亡

Apoptosis of rat liver in cold preservation with custom-designed KYL solution.

作者信息

Li Li, Li Chun-Man, Zhang Bing-Yan, Hu Ming-Dao, Li Xiao-Yan, Ran Jiang-Hua, Huang Ming

机构信息

Department of Surgery, First Hospital of Kunming, Clinical Hospital of Kunming Medical College, Kunming 650011, China.

出版信息

Hepatobiliary Pancreat Dis Int. 2007 Oct;6(5):497-503.

Abstract

BACKGROUND

A suitable perfusate is very important in reducing various problems in liver preservation, prolonging the time of organ preservation and enhancing the quality of donor tissue. University of Wisconsin (UW) solution is the most successful solution for preserving multiple organs at present, but it has many shortcomings. We set out to develop a new liver preservation solution (KYL solution) and study its effects on apoptosis in rat liver undergoing cold preservation.

METHODS

Using non-circulated isolated perfused rat liver (IPRL), we randomly preserved Sprague-Dawley rat livers for 0, 4, 8, 16, 24, and 48 hours with KYL solution or UW solution. The effects were assessed by measuring the content of free radicals in Krebs-Henseleit solution and the intracellular calcium content of hepatocytes, assessing hepatocellular apoptosis and related-gene expression, and observing the morphological changes in liver. To evaluate the protection by KYL and UW solutions in rat liver perfusion and preservation, we chose normal saline for negative comparison.

RESULTS

The intracellular calcium content of the liver preserved in KYL solution was less than that preserved in UW solution. At every different period of preservation, the malonaldehyde and superoxide dismutase content in Krebs-Henseleit solution, the percentage of apoptotic cells and the expression patterns of apoptosis-related-genes were similar in livers preserved in KYL and UW solutions. Morphological changes in the two groups were almost the same. The variables in both groups were better than those of livers preserved in normal saline. Both KYL and UW solutions protected rat liver from ischemia-reperfusion injury.

CONCLUSIONS

KYL solution is superior to UW solution in preventing calcium overload. More severe hepatocyte damage may appear in the KYL group than in the UW group and the effect of KYL solution on apoptosis in rat liver preservation is similar to that of UW solution.

摘要

背景

合适的灌注液对于减少肝脏保存中的各种问题、延长器官保存时间以及提高供体组织质量非常重要。威斯康星大学(UW)溶液是目前保存多种器官最成功的溶液,但它有许多缺点。我们着手开发一种新的肝脏保存溶液(KYL溶液),并研究其对冷保存大鼠肝脏细胞凋亡的影响。

方法

使用非循环离体灌注大鼠肝脏(IPRL),我们用KYL溶液或UW溶液将斯普拉格-道利大鼠肝脏随机保存0、4、8、16、24和48小时。通过测量克氏-亨氏溶液中的自由基含量和肝细胞内钙含量、评估肝细胞凋亡及相关基因表达以及观察肝脏的形态变化来评估效果。为了评估KYL和UW溶液对大鼠肝脏灌注和保存的保护作用,我们选择生理盐水作为阴性对照。

结果

用KYL溶液保存的肝脏细胞内钙含量低于用UW溶液保存的肝脏。在每个不同的保存时期,KYL溶液和UW溶液保存的肝脏中,克氏-亨氏溶液中的丙二醛和超氧化物歧化酶含量、凋亡细胞百分比以及凋亡相关基因的表达模式相似。两组的形态变化几乎相同。两组的各项指标均优于用生理盐水保存的肝脏。KYL和UW溶液均能保护大鼠肝脏免受缺血-再灌注损伤。

结论

KYL溶液在预防钙超载方面优于UW溶液。KYL组可能比UW组出现更严重的肝细胞损伤,且KYL溶液对大鼠肝脏保存中细胞凋亡的影响与UW溶液相似。

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