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Infection and Immunity, April 1999, p. 1633-1639, Vol. 67, No. 4
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Lipopolysaccharide Enhances the Production of Vascular Endothelial Growth Factor by Human Pulp Cells in Culture

K. Matsushita,1 R. Motani,1 T. Sakuta,1 S. Nagaoka,1 T. Matsuyama,2 K. Abeyama,3 I. Maruyama,3 H. Takada,4 and M. Torii1,*

Department of Operative Dentistry and Endodontology, Kagoshima University Dental School, Kagoshima 890-8544,1 Department of Immunology and Medical Zoology2 and Department of Clinical Laboratory Medicine,3 Kagoshima University Medical School, Kagoshima 890-8520, and Department of Microbiology and Immunology, Tohoku University School of Dentistry, Sendai 980-8575,4 Japan

Received 8 September 1998/Returned for modification 2 November 1998/Accepted 19 January 1999

We investigated whether vascular endothelial growth factor (VEGF) production by human pulp cells (HPC) is regulated by lipopolysaccharide (LPS) in relation to the pathogenesis of pulpitis. Although HPC incubated with medium alone only marginally expressed VEGF mRNA and produced a low level of VEGF as detected by enzyme-linked immunosorbent assay, the VEGF mRNA expression and VEGF production were markedly enhanced upon stimulation with LPS from Escherichia coli. Prevotella intermedia LPS, phorbol 12-myristate 13-acetate, and interleukin-6 also induced VEGF mRNA expression in HPC. A simian virus 40-infected HPC line also exhibited increased VEGF mRNA expression in response to E. coli LPS, but lung and skin fibroblasts did not. Fetal bovine serum (FBS) increased the sensitivity of HPC to LPS in a dose-dependent manner. HPC did not express membrane CD14 on their surfaces. However, the anti-CD14 monoclonal antibody MY4 inhibited VEGF induction upon stimulation with LPS in HPC cultures in the presence of 10% FBS but not in the absence of FBS. LPS augmented the VEGF production in HPC cultures in the presence of recombinant human soluble CD14 (sCD14). To clarify the mechanisms of VEGF induction by LPS, we examined the possible activation of the transcription factor AP-1 in HPC stimulated with LPS, by a gel mobility shift assay. AP-1 activation in HPC was clearly observed, whereas that in skin fibroblasts was not. The AP-1 inhibitor curcumin strongly inhibited LPS-induced VEGF production in HPC cultures. In addition, a protein synthesis inhibitor, cycloheximide, inhibited VEGF mRNA accumulation in response to LPS. These results suggest that the enhanced production of VEGF in HPC induced by LPS takes place via an sCD14-dependent pathway which requires new protein synthesis and is mediated in part through AP-1 activation.

* Corresponding author. Mailing address: Department of Operative Dentistry and Endodontology, Kagoshima University Dental School, 8-35-1 Sakuragaoka, Kagoshima 890-8544, Japan. Phone and fax: 81-99-275-6190. E-mail: toriim{at}dentb.hal.kagoshima-u.ac.jp.

Infection and Immunity, April 1999, p. 1633-1639, Vol. 67, No. 4
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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