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-80- Pharmacology 9/4 Room-D
BIOCHEMICAL PROPERTIES OF A MINT ALLERGEN.
Eishin Morita, Michihiro Hide, Shoso Yamamoto.
Department of Dermatology, Hiroshima University School of Medicine, Hiroshima, Japan.
Although in recent years, our knowledge of the structure of plant antigens has increased, a mint allergen which causes contact dernatitis has not been characterized. We experienced a 50-years old woman who showed contact hypersensitivity to several mint leaves and analyzed biochemical properties of an allergen extracted from homogenized mint leaves. Upon Sephacryl S200 gel-chromatography followed by reversed phase-C18-HPLC, the mint allergen was purified in a single peak. The allergen was heat stable (100oC, for 15min) and showed maximum UV absorption band at 271 nm, indicating lipid-like properties of the allergen.
-81- Pharmacology 9/4 Room-D
A STUDY OF EFFECT OF AZELASTIN ON GROWTH OF CULTURED NORMAL FIBROBLAST AND ITS PRODUCTION OF CYTOKINES.
Seiji Maeshima , Toru Minami , Hidenari Kusakabe , Kimihiro Kiyokane.
Department of Dermatorogy , Osaka Medical Collage , Osaka , Japan.
Purpose: A study of effect of Azelastin on growth of cultured normal fibroblasts and production of cytokines. Methord: NHDF were plated at a density of 2500 cells / well, and after 24 hrs culture were treated with 0¡Á5¡ß10-4 M of Azelastin. The number of cells were counted by modified MTT ( WST-1 ) assay methord after 24 ,48 , 72 , 96 hrs incubation respectively. The culture supernatant were assayed after 72 hrs incubation with Azelastin using IL1¦Â, TNF¦Á, IFN¦Ã, TGF¦Â1, bFGF, and procollage ELISA kits. Result: Over 1¡ß10-5 M of Azelastin inhibited cell proliferation. Production of IL1¦Â, TNF¦Á, IFN¦Ã was not observed. 1¡ß10-4 M , 5 ¡ß10-5 M of Azelastin stimulated IL6 production. Over 1¡ß10-5 M of Azelastin suppressed production of TGF¦Â1 , bFGF, procollagen. Conclusion: Over 1¡ß10-5 M of Azelastin downregulated proliferation of normal fibroblasts , procollagen , and cytokines ( TGF¦Â1 , bFGF ) through cell toxicity. Overexpression of IL6 is still studied .
-82- Pharmacology 9/4 Room-D
MODULATION OF CELL ADHESION MOLECULES BY GRISEOFULVIN.
Akihiko Asahina, Kunihiko Tamaki.
Department of Dermatology, University of Tokyo School of Medicine, Tokyo, Japan.
Griseofulvin (GF), a widely used antifungal agent, is also useful in the therapy of several inflammatory skin diseases. In this study, human peripheral blood cells, human dermal microvascular endothelial cells (HDMEC), and human umbilical vein endothelial cells (HUVEC) were incubated with or without the presence of GF to analyze the modulation of cell adhesion molecules. By flowcytometry, the expression of L-selectin on neutrophils was diminished by GF dose-dependently (10-100 mg/ml) while that on lymphocytes remained unchanged. The induction of Mac-1 on neutrophils was not affected. On both TNFa-stimulated HDMEC and IL-1a-stimulated HUVEC, GF suppressed the expression of VCAM-1 although it had little effect on ICAM-1. Moreover, the particles of E-selectin, visualized by fluorescent microscopy, formed clumps by the addition of GF. These results reveal the immunomodulatory potential of GF and suggest its new clinical application.
-83- Pharmacology 9/4 Room-D
EFFECT OF NADIFLOXACIN ON POLY-MORPHONUCLEAR LEUKOCYTE FUNCTIONS AND BACTERIAL LIPASE ACTIVITY.
Noriko Murayama1, Kenji Takamori1, Akimasa Someya2, Isao Nagaoka2.
1Department of Dermatology, Juntendo University Urayasu Hospital, Urayasu, Chiba, 2Department of 2nd Biochemistry, Jundtendo University School of Medicine, Tokyo, Japan.
To clarify the pharmacological action of nadifloxacin (NDFX) on acne formation, the effect of NDFX on polymorphonuclear leukocyte (PMN) func-tion (chemotaxis, lysosomal enzyme release, O£²¡Ý formation) and bacteriallipase activity was investigated. Chemotaxis was measured by the Boydenchamber method. O£²¡Ý formation was determined by measuring the reductionof cytochrome C. (1) NDFX and minocycline (MINO) inhibited PMN chemo- taxis depending on concentration. (2) NDFX and MINO did not affect the lysosomal enzyme release from PMNs. (3) NDFX and MINO suppresed O£²¡Ýformation concentration dependently. These findings suggest that NDFX inhibits acne formation by suppressing PMN chemotaxis and superoxide formation, and through bactericidal action.
-84- Pharmacology 9/4 Room-D
EFFECT OF NICOTINE ON HEME OXYGENASE-1 AND INFLAMMATORY CYTOKINE GENE EXPRESSION IN FIBROBLASTS.
Atsushi Takamiyagi, Shigeru Sassa.
Laboratory for Biochemical Hematology, The Rockefeller University, New York, USA.
Transdermal nicotine patch has been used for treatment of ulcerative colitis, though its mechanism of action is unclear. In order to examine whether it may have an anti-inflammatory effect, we determined the effect of nicotine treatment of cultured human fibroblasts on the expression of a variety of inflammatory markers, including mRNAs encoding microsomal heme oxygenase-1 (HO-1), the rate- limiting enzyme of heme catabolism and a acute-phase reactant, IL-6, IL-6 receptor (IL-6R), and IL-1¦Â. In fibroblasts derived from infant foreskin, HO-1 mRNA levels decreased by the treatment with 40 mM of nicotine or at higher concentrations. IL-6 mRNA levels also decreased by nicotine treatment, but not IL-6R mRNA which increased by the treatment. IL-1¦Â mRNA levels were examined by treatment of cells with TNF-¦Á, with or without nicotine. IL-1¦ÂmRNA levels in cells treated with TNF-¦Á significantly increased by the presence, compared with the absence of nicotine. These findings indicated that nicotine treatment suppresses of certain inflammatory proteins, such as HO-1 and IL-6 mRNA, and suggest that it may be involved in the downregulation of inflammatory processes.