Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 387, Issue 9, pp 861–871 | Cite as

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

  • Zhong-Bao Yang
  • Bin Tan
  • Ting-Bo Li
  • Zheng Lou
  • Jun-Lin Jiang
  • Ying-Jun Zhou
  • Jie Yang
  • Xiu-Ju Luo
  • Jun Peng
Original Article


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.


Hypoxia/reoxygenation NADPH oxidase PC12 cell vitexin Compound B-1 Oxidative injury 



This work was supported by the National Nature Science Foundation of China (No. 81373409 to Jun Peng; No. 81373408 to Jun-Lin Jiang), Hunan Provincial Natural Science Foundation of China (No. 13JJ2008 to Jun Peng), and Doctoral Fund of Ministry of Education of China (No. 20120162110056 to Jun Peng). We sincerely thank Dr. Qing-Jie Li (Department of Internal Medicine, The University of Texas Medical Branch, USA) for his kind assistance in fixing the language problems in our manuscript.

Supplementary material

210_2014_1006_MOESM1_ESM.doc (414 kb)
Supplemental Fig. 1 (DOC 414 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Zhong-Bao Yang
    • 1
  • Bin Tan
    • 1
  • Ting-Bo Li
    • 1
  • Zheng Lou
    • 1
  • Jun-Lin Jiang
    • 1
  • Ying-Jun Zhou
    • 2
  • Jie Yang
    • 3
  • Xiu-Ju Luo
    • 4
  • Jun Peng
    • 1
  1. 1.Department of Pharmacology, School of Pharmaceutical SciencesCentral South UniversityChangshaChina
  2. 2.Department of Medicinal Chemistry, School of Pharmaceutical SciencesCentral South UniversityChangshaChina
  3. 3.Department of NeurologyXiangya Hospital, Central South UniversityChangshaChina
  4. 4.Department of Laboratory MedicineXiangya School of Medicine, Central South UniversityChangshaChina

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