New inhibitors for expression of IgE receptor on human mast cell

Bioorganic and Medicinal Chemistry Letters

Volumn 20, Issue 7, 2010, Pages 2299-2302

  Tamura, Satoru ^a   Yoshihira, Kunichika ^a   Fujiwara, Katsuaki ^a   Murakami, Nobutoshi ^a  

^a OSAKA UNIVERSITY   (Japan)

Author keywords

Anti allergy; Delphinidin; Epigallocatechin gallate; Fc RI; Human mast cell; Tricetinidin

Indexed keywords

ANTIALLERGIC AGENT; APIGENIN; BAICALEIN; CATECHIN GALLATE; CYANIDIN; DELPHINIDIN; EPICATECHIN; EPICATECHIN GALLATE; EPIGALLOCATECHIN; EPIGALLOCATECHIN GALLATE; GALLOCATECHIN GALLATE; IMMUNOGLOBULIN E RECEPTOR; LUTEOLIN; MALVIDIN CHLORIDE; NORWOGONIN; ORIENTIN; PEONIDIN; SCUTELLAREIN; TRICETINIDIN; UNCLASSIFIED DRUG; WOGONIN;

ARTICLE; BIOLOGICAL ACTIVITY; CELL SURFACE; CONTROLLED STUDY; DRUG MECHANISM; DRUG POTENCY; FLOW CYTOMETRY; FLUORESCENT ANTIBODY TECHNIQUE; HUMAN; HUMAN CELL; MAST CELL; PROTEIN EXPRESSION; STRUCTURE ACTIVITY RELATION;

ANTHOCYANINS; ANTI-ALLERGIC AGENTS; CATECHIN; CELL LINE; FLAVONOIDS; GENE EXPRESSION; HUMANS; HYPERSENSITIVITY; MAST CELLS; RECEPTORS, IGE;

EID: 77949486905     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2010.01.160     Document Type: Article

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3. 3 twice, the cells were treated with anti-human FcεRI α-chain antibody (0.001 μg, Kyokuto) on ice for 60 min. Then the cells were rinsed with the PBS twice and incubated with FITC labeled anti-mouse IgG antibody (0.001 μg, Cosmo Bio) on ice for 45 min in the dark. After duplicate washing with the PBS, the harvested cells were analyzed by flow cytometry (FACScalibur, Becton Dickinson). All samples were assessed for inhibitory activity for FcεRI expression in triplicate. In the cases of co-incubation with only DMF and treatment with only FITC labeled anti-mouse IgG antibody, the fluorescent scores were indicated as mean(control) and mean(back), respectively. Inhibition ratio of each sample was determined by the following equation:Inhibition ratio (%) = 100 × [mean(control) - mean(sample)/mean(control) - mean(back)].
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9. 2 for 36 h. Total cellular RNA was isolated using QuickPrep micro mRNA Purification Kit (Amersham) according to the manufacturer's instructions. First strand cDNA was synthesized using anchored oligo(dT) primers and ReverTra Ace reverse transcriptase (TOYOBO). The resultant cDNAs were respectively amplified by using the following primers: FcεRIα forward, 5′ataaaagctccgcgtgagaa3′, reverse, 5′tccttgagcacagacgtttc3′; FcεRIβ forward, 5′ttaccaggacctctaggagtgg3′, reverse 5′aggctggatgaaaaggtgtt3′; FcεRIγ forward 5′ccagcagtggtcttgctcttact3′, reverse 5′gcatgcaggcatatgtgatgcca3′; G3PDH forward 5′gatgacatcaagaaggtggtg3′, reverse 5′gctgtagccaaattcgttgtc3′. The amplified PCR products were subjected to electrophoresis on a 1.5% TAE (Tris-acetate EDTA buffer) agarose gel, then stained with ethidium bromide. Image analysis for each blot was conducted by Scion image (Scion) to determine amount of the expressed mRNA and quantification of each mRNA was carried out in triplicate. G3PDH was used as a control to correct expression of FcεRIα, β, and γ. The relative expression rates of the three subunits were described as compared with that of unstimulated HMC-1 cells.