Uptake of bacterial lipopolysaccharide and expression of tumor necrosis factor-&alpha;-mRNA in isolated rat intrahepatic bile duct epithelial cells

doi:10.3748/wjg.v3.i1.3

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Mar 15, 1997 (publication date) through Nov 5, 2024

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

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World J Gastroenterol. Mar 15, 1997; 3(1): 3-5
Published online Mar 15, 1997. doi: 10.3748/wjg.v3.i1.3

Uptake of bacterial lipopolysaccharide and expression of tumor necrosis factor-α-mRNA in isolated rat intrahepatic bile duct epithelial cells

Xian-Ming Chen, De-Wu Han, K Noguchi, K Tanikawa

Xian-Ming Chen, De-Wu Han, Institute of Hepatology, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China

K Noguchi, K Tanikawa, Second Department of Medicine, Kurume University School of Medicine, Kurume, Japan

Xian-Ming Chen, Associate Professor of pathophysiology with major specialty in hepatic pathophysiology, with 15 published papers

Author contributions: All authors contributed equally to the work.

Supported by The Natural Science Foundation for Youth of Shanxi Province, No. 95013.

Correspondence to: Dr. Xian-Ming Chen, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China

Telephone: +86-351-2021511-521 Fax: +86-351-2024239

Received: January 26, 1996
Revised: September 1, 1996
Accepted: December 1, 1996
Published online: March 15, 1997

Abstract

AIM: To study the uptake of bacterial lipopolysaccharides (LPS) and expression of tumor necrosis factor α-mRNA (TNF-α-mRNA) with cultured rat intrahepatic bile duct epithelial cells.

METHODS: By using fluorescent, immunohistochemical and in situ hybridization techniques, the uptake of Escherichia coli LPS and expression of TNF-α-mRNA with isolated rat intrahepatic bile duct epithelial cells were observed with confocal laser scanning microscopy.

RESULTS: Positive reactions to LPS were found in the cytoplasm of isolated intrahepatic bile duct epithelial cells after incubation with LPS for 15 min and the FITC fluorescent intensity against LPS was significantly higher than that of the controls (121.45 μFI/μm²± 15.62 μFI/μm²vs 32.12 μFI/μm²± 9.64 μFI/μm², P < 0.01). After incubation with LPS for 3 h, fluorescein isocyanate (FITC) fluorescent intensities of the expression of TNF-α-mRNA with fluorescent in situ hybridization in the cytoplasm and nuclei of the cultured bile duct epithelial cells were significantly higher than those of the controls (189.15 μFI/μm²± 21.33 μFI/μm²vs 10.00 μFI/μm²± 8.99 μFI/μm², 64.85 μFI/μm²± 14.99 μFI/μm²vs 21.20 μFI/μm²± 2.04 μFI/μm², respectively (P < 0.01)). The increase of FITC fluorescent intensity of TNF-α-mRNA expression in the cytoplasm peaked at 6 h after incubation (221.38 μFI/μm²± 22.99 μFI/μm²). At various time points after incubation with LPS, the increase of fluorescent intensities of TNF-α-mRNA in the cytoplasm were much higher than those in the nuclei (P < 0.01).

CONCLUSION: LPS can act on and enter into isolated intrahepatic bile duct epithelial cells and stimulate the expression of TNF-α-mRNA.

Keywords: Lipopolysaccharides; Epithelial cell bile ducts; Tumor necrosis factor; In situ hybridization