[阿霉素诱导热休克蛋白72对大鼠肝脏冷缺血再灌注损伤的保护作用]

[The protective effect of heat shock protein 72 by Doxorubicin in cold ischemia-reperfusion injury of the rat liver].

作者信息

Chen Hao, Peng Cheng-hong, Deng Xia-xing, Qiu Wei-hua, Shen Bai-yong, Yang Wei-ping, Li Hong-wei

机构信息

Center of Organ Transplantation, Ruijin Hospital, Medical School of Shanghai Jiaotong University, Shanghai 200025, China.

出版信息

Zhonghua Wai Ke Za Zhi. 2006 Mar 1;44(5):310-3.

PMID:16635388

Abstract

OBJECTIVE

To observe induction of heat shock reaction by pretreatment of Doxorubicin (DXR) in long-term cold preservation-reperfusion injury of the rat liver.

METHODS

The rats were administered intravenously by DXR at a dose of 1 mg/kg body weight in DXR group and by saline in control group. After 48 hours, the rat liver was perfused by using cold University of Wisconsin (UW) solutions and was preserved in UW solution at 4 degrees C for 48 hours. Recipient liver was perfused for 1 and 3 hours after orthotopic liver transplantation. Tumor necrosis factor-alpha (TNF-alpha) mRNA, cytokine-induced neutrophil chemoattractant (CINC) mRNA, macrophage inflammatory protein (MIP-2) mRNA was measured by RT-PCR and heat shock protein 72 (HSP72), nuclear factor-kappaB (NF-kappaB) by Western blot. The serum levels of TNF-alpha, CINC, MIP-2 by ELISA and AST were measured. The survival rate of 7 days was observed.

RESULTS

The expression of TNF-alpha mRNA, CINC mRNA and MIP-2 mRNA was stronger in control group than in DXR group. HSP72 was expressed in SA group but not in control group and oppositely NF-kappaB was expressed in control group but not in DXR group. Serum AST, TNF-alpha, CINC and MIP-2 concentrations were significantly lower in DXR group than in control group (P < 0.05). The survival rate of 7 days was significantly higher in DXR group than in control group (50% vs. 0%, P < 0.05).

CONCLUSIONS

These data suggested that long-term cold ischemia-reperfusion injury was attenuated in liver graft with pretreatment of DXR. The induction of HSP72 may offer protection to hepatocytes by restraining the activation of NF-kappaB and inflammation.

摘要

目的

观察阿霉素（DXR）预处理对大鼠肝脏长期冷保存-再灌注损伤中热休克反应的诱导作用。

方法

DXR组大鼠静脉注射剂量为1mg/kg体重的DXR，对照组注射生理盐水。48小时后，用冷的威斯康星大学（UW）溶液灌注大鼠肝脏，并在4℃的UW溶液中保存48小时。原位肝移植后，对受体肝脏进行1小时和3小时的灌注。通过逆转录聚合酶链反应（RT-PCR）检测肿瘤坏死因子-α（TNF-α）信使核糖核酸（mRNA）、细胞因子诱导的中性粒细胞趋化因子（CINC）mRNA、巨噬细胞炎性蛋白（MIP-2）mRNA，通过蛋白质免疫印迹法检测热休克蛋白72（HSP72）、核因子-κB（NF-κB）。通过酶联免疫吸附测定法（ELISA）检测血清中TNF-α、CINC、MIP-2水平以及天冬氨酸转氨酶（AST）。观察7天的生存率。

结果

对照组中TNF-α mRNA、CINC mRNA和MIP-2 mRNA的表达强于DXR组。HSP72在DXR组中表达而在对照组中不表达，相反，NF-κB在对照组中表达而在DXR组中不表达。DXR组血清AST、TNF-α、CINC和MIP-2浓度显著低于对照组（P<0.05）。DXR组7天生存率显著高于对照组（50%对0%，P<0.05）。