Equol protects PC12 neuronal cells against hypoxia/reoxygenation injury in vitro by reducing reactive oxygen species production.
- Author:
Wei YU
1
;
Xiuling DENG
;
Zhen MA
;
Yan WANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Hypoxia; Cell Survival; Down-Regulation; Equol; pharmacology; L-Lactate Dehydrogenase; metabolism; Malondialdehyde; metabolism; Membrane Glycoproteins; metabolism; NADPH Oxidase 2; NADPH Oxidases; metabolism; Neurons; drug effects; metabolism; Neuroprotective Agents; pharmacology; PC12 Cells; Phosphorylation; Rats; Reactive Oxygen Species; metabolism; src-Family Kinases; metabolism
- From: Journal of Southern Medical University 2016;36(1):1-7
- CountryChina
- Language:English
-
Abstract:
OBJECTIVEBoth of gp91(phox) (an isoform of nicotinamide adenine dinucleotide phosphate-reduced oxidases) and Src (a non-receptor protein tyrosine kinase) play a prominent role in mediating hypoxia/reoxygenation injury of neurons. The present study was designed to investigate the neuroprotective effect of equol, a predominant active metabolite of daidzein, against hypoxia/reoxygenation injury in rat pheochromocytoma cell line (PC12) and explore the underlying mechanisms.
METHODSPC12 cells exposed to hypoxia/reoxygenation injury were examined for reactive oxygen species (ROS) using dihydroethidium and 2', 7'-dichlorofluorescein diacetate and analyzed for changes in lactate dehydrogenase (LDH) activity and malondialdehyde (MDA) content. The expression levels of gp91(phox) and phosphorylated Src-Tyr416 (p-Src) were measured using Western blotting.
RESULTSEquol dose-dependently restored the cell viability and decreased LDH activity and MDA content in culture medium of PC12 cells exposed to hypoxia/reoxygenation. Pretreatment of the cells with 10(-5) and 10(-6) mol/L equol inhibited hypoxia/reoxygenation-induced increase of ROS. PC12 cells treated with equol prior to hypoxia/reoxygenation injury showed significant enhancement of the protein levels of gp91(phox) and p-Src.
CONCLUSIONEquol confers neuroprotection against hypoxia/reoxygenation injury in PC12 cells by inhibiting the generation of ROS very likely as a result of down-regulation of gp91(phox) and inhibition of Src phosphorylation.