Differential effects of gadolinium chloride on Kupffer cells in vivo and in vitro
Differential effects of gadolinium chloride on Kupffer cells in vivo and in vitro
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DOI：
10.1016/j.biocel.2003.08.004
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发表时间：
2004-03-01
影响因子：
4
通讯作者：
Olynyk, JK
中科院分区：
文献类型：
--
作者：
Lee, CM;Yeoh, GC;Olynyk, JK
在活体内研究枯否细胞在肝脏疾病中的作用时，常使用氯化GdCl。GdCl对培养的Kupffer细胞的体外作用尚不清楚。本研究的目的是研究大鼠Kupffer细胞在给予GdCl后的肿瘤坏死因子α的产生、吞噬功能以及ED1和ED2抗原的表达。在体内实验中，大鼠注射10 mg/kg GdCl4或无菌生理盐水。于注射GdCl后1~3、5、8d处死动物，处死前4h给予脂多糖3 mg/kg。测定肝组织ED1、ED2阳性巨噬细胞数和肿瘤坏死因子α(TNFpha)基因表达水平。在体外实验中，Kupffer细胞在0-270um GdCl中培养24 h，然后检测细胞存活率、对内毒素(10 ng/ml)反应产生的TNFα蛋白、吞噬功能以及ED1和ED2染色。在体内，注射GdCl后1~5天，ED1阳性细胞比例由87%降至3%，肝组织中肿瘤坏死因子α基因表达水平下降60%(P<；0.01)。在体外，吞噬功能随着GdCl2浓度的增加而降低。GdCl(0~27um)对Kupffer细胞存活率、TNFα产生、ED1或ED2染色均无影响。我们的结论是，GdCl在体内显著降低Kupffer细胞ED2的表达。在体外，GdCl对巨噬细胞的吞噬作用具有剂量依赖性，但仅在高浓度时影响细胞存活率和TNFpha的产生。GdCl对培养的Kupffer细胞的ED2表达无影响。(C)2003爱思唯尔有限公司。保留所有权利。
Gadolinium chloride (GdCl) is commonly used to study the role of Kupffer cells in liver disease in vivo. The in vitro effects of GdCl on cultured Kupffer cells are poorly characterised. The aim of this study was to characterise rat Kupffer cell TNFalpha production, phagocytic function, and ED1 and ED2 antigen expression following the administration of GdCl. For in vivo experiments, rats received 10 mg/kg GdCl IV or sterile saline. Lipopolysaccharide 3 mg/kg IP (LPS) was administered 4 h prior to sacrifice on Days 1-3, 5 or 8 following GdCl injection. Hepatic ED1 and ED2 positive macrophage numbers and TNFalpha mRNA levels were determined. For in vitro experiments, Kupffer cells were cultured in the presence of 0-270 muM GdCl for 24 h following which viability, TNFalpha protein production in response to LPS (10 ng/ml), phagocytosis, and ED1 and ED2 staining were evaluated. In vivo, the proportion of ED1 positive cells which were ED2 positive was reduced from 87 to 3% and hepatic TNFalpha mRNA levels following LPS declined by 60% over Days 1-5 after injection of GdCl (P < 0.01). In vitro, phagocytosis declined with increasing concentrations of GdCl. GdCl (0-27 muM) did not effect cultured Kupffer cell viability, TNFalpha production, ED1 or ED2 staining. We conclude that GdCl significantly reduces ED2 expression by Kupffer cells in vivo. In vitro, GdCl has a dose dependent effect on phagocytosis but only effects viability and TNFalpha production at high concentrations. ED2 expression of cultured Kupffer cells is not affected by GdCl. (C) 2003 Elsevier Ltd. All rights reserved.