Diphenhydramine interferes with MTT reduction assay.
- Author:
Yao SHEN
1
;
Chen-Hui ZHANG
;
Wei-Wei HU
;
Zhong CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Astrocytes; drug effects; metabolism; physiology; Cell Survival; Cells, Cultured; Diphenhydramine; pharmacology; Drug Interactions; Formazans; pharmacokinetics; Rats; Rats, Sprague-Dawley; Tetrazolium Salts; pharmacokinetics
- From: Journal of Zhejiang University. Medical sciences 2011;40(2):163-168
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo determine the effects of organic amine diphenhydramine on the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide dye (MTT) reduction assay.
METHODSThe primarily cultured cortical astrocytes were incubated with various concentrations of diphenhydramine for 24 h. To analyze the effects of diphenhydramine and other organic amines on the MTT assay, the data obtained from the MTT assay were compared with the results obtained from morphological observation and hoechst 33342 and propidium iodide (PI) nucleus double staining.
RESULTThe MTT assay showed that diphenhydramine (10(-4)mol/L), pyrilamine (10 (-4)mol/L) and zolantidine (10 (-5)mol/L) caused a significant increase in MTT reduction in astrocytes. However there was no proliferation, apoptosis or necrosis detected by hoechst and PI nucleus double staining. Light microscopy revealed that exocytosis of formazan granules was inhibited by diphenhydramine.
CONCLUSIONDiphenhydramine and other organic amines may enhance MTT reduction by suppression of MTT formazan exocytosis in astrocytes, which may affect the results of cell viability studies.
