Protective effect of vitexin compound B-1 against hypoxia/reoxygenation-induced injury in differentiated PC12 cells via NADPH oxidase inhibition.

Abstract

Vitexin compound B-1 (VB-1) is a novel member of the vitexins family isolated from the seeds of the Chinese herb Vitex negundo. This study aims to investigate whether VB-1 is able to protect nerve cells against oxidative injury and whether the antioxidative effects of VB-1 occur through a mechanism involving the inhibition of NADPH oxidase (NOX) in a manner of hypoxia-inducible factor 1α (HIF-1α)-dependent. To establish a neuronal in vitro model of oxidative stress, the differentiated PC12 cells were subjected to 5 h of hypoxia followed by 20 h of reoxygenation (H/R). Three dosages of VB-1 (10(-8), 10(-7), and 10(-6) M) were chosen to evaluate the effect of VB-1 on H/R-induced injury and the underlying mechanisms. At the end of the experiments, culture mediums and cells were collected for analysis of cellular apoptosis, lactate dehydrogenase (LDH) and caspase 3/7-like activities, reactive oxygen species (ROS) levels, 4-hydroxynonenal (4-HNE) and malondialdehye (MDA) contents, and HIF-1α and NOX expression, respectively. Our results showed that cell injury (indicated by apoptosis ratio, caspase 3/7-like activity, and LDH release), oxidative stress (indicated by ROS production, 4-HNE, and MDA contents), NOX activity, and NOX expression (NOX2 and NOX4 isoforms) were dramatically increased in PC12 cells following H/R, which were attenuated in the presence of VB-1 at dosage of 10(-7) or 10(-6) M. There was no significant change in HIF-1α expression in all experimental groups. These results provide evidence that VB-1 is able to protect the PC12 cells against H/R-induced injury through a mechanism involving the suppression of NOX expression and subsequent reduction of ROS production. The effect of VB-1 on H/R-induced NOX expression is independent on HIF-1α inhibition.

• Authors: Zhong-Bao Yang, Bin Tan, Ting-Bo Li, Zheng Lou, Jun-Lin Jiang, Ying-Jun Zhou, Jie Yang, Xiu-Ju Luo, Jun Peng
• Journal: Naunyn-Schmiedeberg's archives of pharmacology (Naunyn Schmiedebergs Arch Pharmacol) Vol. 387 Issue 9 Pg. 861-71 (Sep 2014) ISSN: 1432-1912 [Electronic] Germany
• PMID: 24947869 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 6-hydroxy-4-(4-hydroxy-3-methoxyphenyl)-3-hydroxymethyl-7-methoxy-3,4-dihydro-2-naphthaldehyde
• Aldehydes
• Lignans
• Neuroprotective Agents
• RNA, Messenger
• Reactive Oxygen Species
• Malondialdehyde
• NADPH Oxidases
• Casp3 protein, rat
• Caspase 3
• Caspase 7
• 4-hydroxy-2-nonenal

Topics

• Aldehydes (metabolism, pharmacology)
• Animals
• Apoptosis (drug effects)
• Caspase 3 (metabolism)
• Caspase 7 (metabolism)
• Cell Differentiation
• Hypoxia (metabolism)
• Lignans (pharmacology)
• Malondialdehyde (metabolism)
• NADPH Oxidases (antagonists & inhibitors, genetics, metabolism)
• Neuroprotective Agents (pharmacology)
• PC12 Cells
• RNA, Messenger (metabolism)
• Rats
• Reactive Oxygen Species (metabolism)