沙利度胺对内毒素诱导的肝损伤的保护作用。

Protective effect of thalidomide on endotoxin-induced liver injury.

作者信息

Enomoto Nobuyuki, Takei Yoshiyuki, Hirose Miyoko, Kitamura Tsuneo, Ikejima Kenichi, Sato Nobuhiro

机构信息

Department of Gastroenterology, Juntendo University School of Medicine, Tokyo, Japan.

出版信息

Alcohol Clin Exp Res. 2003 Aug;27(8 Suppl):2S-6S. doi: 10.1097/01.ALC.0000078606.59842.01.

DOI:10.1097/01.ALC.0000078606.59842.01

PMID:12960498

Abstract

BACKGROUND

Activation of Kupffer cells by lipopolysaccharide (LPS) plays a pivotal role in the onset of pathophysiological events that occur during endotoxemia, and intracellular calcium concentration ([Ca2+]i) is involved in LPS-stimulated cytokine production. Tumor necrosis factor (TNF)-alpha is produced exclusively by the monocyte-macrophage lineage, which is mostly made up of Kupffer cells, and thalidomide has been shown to reduce TNF-alpha production from macrophages. However, there is increasing evidence that TNF-alpha may play a role in the initiation or progression of multiple organ failure syndrome. Therefore, the purpose of this work was to determine whether thalidomide could prevent LPS-induced liver injury.

METHODS

Rats were given a single oral dose of thalidomide (5 mg/kg). To assess the sensitization of Kupffer cells, LPS (5 or 10 mg/kg) was administered intravenously, and mortality, liver histology, and transaminases were evaluated 24 hr later. Kupffer cells were isolated 2 hr after thalidomide treatment. After the addition of LPS, [Ca2+]i was measured by using a microspectrofluorometer with the fluorescent indicator fura-2, and TNF-alpha was measured by enzyme-linked immunosorbent assay.

RESULTS

LPS caused focal necrosis with neutrophil infiltration in the liver. Moreover, LPS dramatically increased transaminases. These pathologic parameters and increases of serum transaminases were diminished markedly by thalidomide. In isolated Kupffer cells, LPS-induced increases in [Ca2+]i and TNF-alpha production were suppressed by treatment with thalidomide. To further explore the mechanism by which thalidomide directly abrogated Kupffer cell sensitivity to LPS, we determined the effect of thalidomide (5 microM) in vitro on LPS-induced [Ca2+]i response and TNF-alpha production. With the addition of thalidomide (5 microM) in vitro to the culture media for 2 hr before LPS, these parameters were suppressed.

CONCLUSIONS

Thalidomide prevents LPS-induced liver injury via mechanisms dependent on the suppression of TNF-alpha production from Kupffer cells.

摘要

背景

脂多糖（LPS）激活库普弗细胞在内毒素血症期间发生的病理生理事件的起始过程中起关键作用，并且细胞内钙浓度（[Ca2+]i）参与LPS刺激的细胞因子产生。肿瘤坏死因子（TNF）-α仅由单核细胞-巨噬细胞谱系产生，而该谱系主要由库普弗细胞组成，并且沙利度胺已被证明可减少巨噬细胞产生TNF-α。然而，越来越多的证据表明TNF-α可能在多器官功能衰竭综合征的起始或进展中起作用。因此，本研究的目的是确定沙利度胺是否可以预防LPS诱导的肝损伤。

方法

给大鼠单次口服沙利度胺（5mg/kg）。为了评估库普弗细胞的敏感性，静脉注射LPS（5或10mg/kg），并在24小时后评估死亡率、肝脏组织学和转氨酶。在沙利度胺治疗2小时后分离库普弗细胞。加入LPS后，使用带有荧光指示剂fura-2的显微分光荧光计测量[Ca2+]i，并通过酶联免疫吸附测定法测量TNF-α。

结果

LPS导致肝脏出现局灶性坏死并伴有中性粒细胞浸润。此外,LPS显著增加转氨酶。这些病理参数和血清转氨酶的升高被沙利度胺显著减轻。在分离的库普弗细胞中，沙利度胺处理可抑制LPS诱导的[Ca2+]i增加和TNF-α产生。为了进一步探讨沙利度胺直接消除库普弗细胞对LPS敏感性的机制，我们在体外确定了沙利度胺（5μM）对LPS诱导的[Ca2+]i反应和TNF-α产生的影响。在LPS加入前2小时，在体外向培养基中添加沙利度胺（5μM），这些参数受到抑制。