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半肝血流阻断门静脉结扎术中再灌注诱导的肝细胞凋亡减弱与 bcl-2/bax 比值的逆转相关。

Attenuation of reperfusion-induced hepatocyte apoptosis is associated with reversed bcl-2/bax ratio in hemi-hepatic artery-preserved portal occlusion.

机构信息

Department of Hepatobiliary Surgery, Shengjing Hospital, China Medical University, Shenyang, China.

出版信息

J Surg Res. 2012 May 15;174(2):298-304. doi: 10.1016/j.jss.2010.12.030. Epub 2011 Jan 19.

Abstract

BACKGROUND

This study aimed to examine the hepatocyte apoptosis in a hepatic blood inflow occlusion rat model without hemi-hepatic arterial control and its association with the expressions of the apoptosis-regulating genes bcl-2 and bax.

MATERIALS AND METHODS

Wistar rats were equally and randomly assigned to undergo sham operation (control group, n = 8), Pringle's maneuver (group PR, n = 32), hemi-hepatic occlusion (group HH, n = 32), or hemi-hepatic artery-preserved portal occlusion (group HP, n = 32). The hepatic blood inflow was interrupted for 30 min using a microvascular clip in the three experimental groups. The clips were removed to achieve hepatic reperfusion for up to 24 h. Blood samples and liver specimens were collected following reperfusion to perform pathologic examination, serum transferase assay, apoptosis analysis, and determination of bcl-2 and bax mRNA and protein expressions.

RESULTS

The reperfusion-related hepatocytic injuries were more severe in the PR group than in the HH and HP groups, both pathologically and biochemically. More reperfused hepatocytes became apoptotic in the PR group than in the HH and HP groups. However, the values of the HH and HP groups were comparable in cellularity, levels of serum transferases, and apoptosis rate following reperfusion. The ratios of bcl-2/bax were reversed, which was more evident in the HH and HP groups than in the PR group.

CONCLUSION

Hemi-hepatic artery-preserved portal occlusion had little effect on hepatocyte apoptosis compared with Pringle's maneuver and caused minor ischemia-reperfusion injury as shown by the reversed bcl-2/bax ratio.

摘要

背景

本研究旨在观察无半肝动脉控制的肝血流阻断大鼠模型中的肝细胞凋亡及其与凋亡调节基因 bcl-2 和 bax 表达的关系。

材料和方法

Wistar 大鼠被平均且随机分为假手术(对照组,n = 8)、Pringle 手法(PR 组,n = 32)、半肝阻断(HH 组,n = 32)或半肝动脉保留门静脉阻断(HP 组,n = 32)。在三组实验中,使用微血管夹阻断肝血流 30 分钟。夹闭移除以实现肝再灌注,最长达 24 小时。再灌注后采集血样和肝组织进行病理检查、血清转氨酶测定、凋亡分析以及 bcl-2 和 bax mRNA 和蛋白表达的测定。

结果

PR 组的再灌注相关肝细胞损伤比 HH 和 HP 组更严重,无论是在病理上还是在生化上。PR 组的再灌注肝细胞凋亡数量多于 HH 和 HP 组。然而,HH 和 HP 组的细胞数量、血清转氨酶水平和再灌注后凋亡率相当。bcl-2/bax 比值发生反转,在 HH 和 HP 组比 PR 组更明显。

结论

与 Pringle 手法相比,半肝动脉保留门静脉阻断对肝细胞凋亡影响较小,bcl-2/bax 比值的反转表明其引起的缺血再灌注损伤较小。

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