氯化钆通过抑制细胞凋亡减轻大鼠肝缺血再灌注损伤。

GdCl3 abates hepatic ischemia-reperfusion injury by inhibiting apoptosis in rats.

机构信息

Department of General Surgery, Shengjing Hospital, China Medical University, Shenyang 110004, China.

出版信息

Hepatobiliary Pancreat Dis Int. 2009 Oct;8(5):518-23.

PMID:19822496

Abstract

BACKGROUND

Gadolinium chloride (GdCl(3)) is a specific inhibitor of Kupffer cells (KCs), which are important promoters of various liver injuries. It is therefore of interest to explore the role of KCs in liver ischemia-reperfusion injury and their relations with apoptosis caused by ischemia-reperfusion injury.

METHODS

One hundred male Wistar rats (190-210 g, 6-7 weeks old) were divided into two groups at random, GdCl(3) group and control group. Samples were collected at 0.5, 1, 6, 12, and 24 hours from each group after reperfusion. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured by an automatic biochemical analyzer. TNF-alpha in serum was measured by enzyme-linked immunosorbent assay (ELISA). Malondialdehyde (MDA) in the liver mitochondria was measured by a colorimetric method. Pathological changes in the liver and immunohistochemical staining for caspase-3 were observed under an optical microscope. The ratio of apoptotic cells was measured by TdT-mediated dUTP nick-end labeling (TUNEL), and ultrastructural features of apoptosis were observed with a transmission electron microscope (TEM).

RESULTS

The levels of ALT in the GdCl(3) group were lower than those in the control group after reperfusion for 0.5, 1, 6 and 12 hours (P<0.05); and the levels of AST in the GdCl(3) group were lower than those in the control group after reperfusion for 6 and 12 hours (P<0.05). The levels of TNF-alpha in the GdCl(3) group were lower than those in the control group after reperfusion for each time (P<0.05). The concentrations of MDA after reperfusion in the GdCl(3) group were lower than those in the control group after reperfusion for 6, 12 and 24 hours (P<0.05). After reperfusion for 0.5, 1, 6 and 12 hours, the integral optical density (IOD) of caspase-3-positive cells was lower in the GdCl(3) group than in the control group (P<0.05). After reperfusion for 1, 6, and 12 hours, the IOD of cells stained by TUNEL in the GdCl(3) group was lower than that in the control group (P<0.05).

CONCLUSIONS

GdCl(3) inhibits the activity of ALT, AST and TNF-alpha, decreases the accumulation of MDA in mitochondria, and depresses the expression of caspase-3 in liver after ischemia-reperfusion. This may be an important protective mechanism by depressing KCs and indirectly inhibiting liver cell apoptosis.

摘要

背景

氯化钆（GdCl(3)）是库普弗细胞（KCs）的特异性抑制剂，KCs 是各种肝损伤的重要促进剂。因此，探讨 KCs 在肝缺血再灌注损伤中的作用及其与缺血再灌注损伤引起的细胞凋亡的关系具有重要意义。

方法

将 100 只雄性 Wistar 大鼠（190-210g，6-7 周龄）随机分为两组，GdCl(3)组和对照组。每组再灌注后 0.5、1、6、12 和 24 小时分别采集样本。采用自动生化分析仪检测血清丙氨酸氨基转移酶（ALT）和天门冬氨酸氨基转移酶（AST）。采用酶联免疫吸附试验（ELISA）检测血清 TNF-α。采用比色法检测肝线粒体丙二醛（MDA）。在光学显微镜下观察肝组织病理学变化和 caspase-3 免疫组化染色。采用末端转移酶介导的 dUTP 缺口末端标记法（TUNEL）测定凋亡细胞的比例，并通过透射电子显微镜（TEM）观察凋亡的超微结构特征。

结果

再灌注 0.5、1、6 和 12 小时后，GdCl(3)组 ALT 水平低于对照组（P<0.05）；再灌注 6 和 12 小时后，GdCl(3)组 AST 水平低于对照组（P<0.05）。再灌注后各时间点，GdCl(3)组 TNF-α水平均低于对照组（P<0.05）。再灌注 6、12 和 24 小时后，GdCl(3)组 MDA 浓度低于对照组（P<0.05）。再灌注 0.5、1、6 和 12 小时后，GdCl(3)组 caspase-3 阳性细胞的积分光密度（IOD）低于对照组（P<0.05）。再灌注 1、6 和 12 小时后，GdCl(3)组 TUNEL 染色细胞的 IOD 低于对照组（P<0.05）。