Basic Research
Copyright ©The Author(s) 2002. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Feb 15, 2002; 8(1): 124-127
Published online Feb 15, 2002. doi: 10.3748/wjg.v8.i1.124
Lipopolysaccharide induced synthesis of CD14 proteins and its gene expression in hepatocytes during endotoxemia
Sheng-Wei Li, Jian-Ping Gong, Chuan-Xin Wu, Yu-Jun Shi, Chang-An Liu
Sheng-Wei Li, Jian-Ping Gong, Chuan-Xin Wu, Yu-Jun Shi, Chang-An Liu, Department of General Surgery, The Second College of Clinical Medicine & the Second Affiliated Hospital of Chongqing University of Medical Sciences, Chongqing 400010, China
Author contributions: All authors contributed equally to the work.
Supported by the National Natural Science Foundation of China (No. 39970719)
Correspondence to: Sheng-Wen Li, Department of General Surgery, The Second College of Clinical Medicine & the Second Affiliated Hospital of Chongqing Medical University, 74 Linjiang Road, Central District, Chongqing 400010, China. lswgg@163.con
Telephone: +86-23-63849075-2100
Received: August 23, 2001
Revised: September 1, 2001
Accepted: September 5, 2001
Published online: February 15, 2002
Abstract

AIM: To observe synthesis of CD14 protein and expression of CD14 mRNA in hepatic tissue and hepatocytes of rats during endotoxemia.

METHODS: The endotoxemia model of Wistar rat was established by injection of a dose of lipopolysaccharide (LPS) (5 mg·kg-1, Escherichia coli O111:B4) via the tail vein, and then the rats were sacrificed after 3, 6, 12 and 24 h in batches. Hepatocytes were isolated from normal and LPS-injected rats by in situ collagenase perfusion technique and were collected to measure the expression of CD14 mRNA and synthesis of CD14 protein by reverse transcript-polymerase chain reaction (RT-PCR) or Western blot analysis. The binding of fluorescein isothiocyanate (FITC)-CD14 polyclonal antibody to isolated hepatocytes was also assessed by flow cytometric analysis (FCM).

RESULTS: In the rats with endotoxemia, the expressions of CD14 mRNA in hepatic tissue and isolated hepatocytes were stronger at 3, 6, and 12 h than that in control rats (3.48 ± 0.15, 5.89 ± 0.62, 4.33 ± 0.18, vs 1.35 ± 0.14 in hepatic tissue, P < 0.01; 4.12 ± 0.17, 6.24 ± 0.64, 4.35 ± 0.18, vs 1.87 ± 0.15 in hepatocytoes, P < 0.01).The synthesis of CD14 protein in hepatic tissue and isolated hepatocytes increases also obviously in 6 and 12 h when compared to that in control rats (13.27 ± 1.27, 17.32 ± 1.35, 11.42 ± 1.20,vs 7.34 ± 0.72 in hepatic tissue, P < 0.01; 14.68 ± 1.30, 17.95 ± 1.34, 11.65 ± 1.19, vs 7.91 ± 0.70 in hepatocytes, P < 0.01). FCM showed that mean fluorescence intensity (MFI) and numbers of FITC-CD14 positive cells in the rats with endotoxemia increased obviously at 3, 6, 12 and 24 h when compared with normal control group (43.4%, 70.2%, 91.4%, 32.6% vs 4.5%, P < 0.01).

CONCLUSION: LPS can markedly promote the synthesis of CD14 protein and up-regulate the expression of CD14 mRNA in isolated hepatocytes and hepatic tissue. Liver might be a main source for soluble CD14 production during endotoxemia.

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