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Cytotechnology

, Volume 63, Issue 2, pp 181–190 | Cite as

Microarray analysis of immediate-type allergy in KU812 cells in response to fulvic acid

  • Hideko Motojima
  • Myra O. Villareal
  • Junkyu Han
  • Hiroko IsodaEmail author
JAACT Special Issue

Abstract

Fulvic acid (FA) is class of compounds of humic substances formed through the degradation of organic substances by chemical and biological processes. FA has been utilized in traditional Chinese medicine and possesses various pharmacological properties. Previously, we reported that FA extracted from solubilized excess sludge (SS-FA) had an inhibitory effect on β-hexosaminidase release in human leukemia basophilic (KU812) cells. In this study, we investigated the effects of SS-FA on the immediate-type allergic reaction and studied its possible mechanisms of action in KU812 cells following activation with phorbol myristate acetate (20 nmol L−1) plus calcium ionophore A23187 (1 μmol L−1) (PMACI). The inhibitory effect of SS-FA on degranulation in PMACI-stimulated KU812 cells was examined using histamine release assay. SS-FA significantly decreased the histamine release in KU812 cells at concentrations of 0.1–10.0 μg mL−1. To gain more information regarding the mechanism of the suppression of degranulation following SS-FA treatment, microarray was conducted to determine which genes were differentially expressed in response to SS-FA in PMACI-activated KU812 cells. From a total of 201 genes in the DNA chip, 28 genes were up-regulated and 173 genes were down-regulated in cells pretreated with SS-FA for 15 min and stimulated with PMACI. From the 71 genes that showed more than two fold change in expression, 16 genes were significantly down-regulated that were subjected to hierarchical clustering. SS-FA affected the expression of genes that were involved in the following pathways: signal transduction, cytokine–cytokine receptor interaction, immune response, cell adhesion molecules and IgE receptor β subunit response.

Keywords

Immediate-type allergy Histamine release Microarray analysis Fulvic acid Solubilized excess sludge 

Abbreviations

DNP-BSA

Dinitrophenylated bovine serum albumin

FA

Fulvic acid

FBS

Fetal bovine serum

KU812

Human leukemia basophilic

PMA

Phorbol 12-myristate 13-acetate

PMACI

PMA plus calcium ionophore A23187

RBL-2H3

Rat basophilic leukemia

SLE

Systemic lupus erythematosus

SS-FA

Fulvic acid extracted from solubilized sludge

Notes

Acknowledgments

This research was partially supported by the Mitsui & Co., Ltd. Environment Fund.

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Hideko Motojima
    • 1
  • Myra O. Villareal
    • 1
  • Junkyu Han
    • 1
  • Hiroko Isoda
    • 1
    Email author
  1. 1.Graduate School of Life and Environmental SciencesUniversity of TsukubaTsukubaJapan

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