Neuroprotective effect of osthole on MPP+-induced cytotoxicity in PC12 cells via inhibition of mitochondrial dysfunction and ROS production

Neurochem Int. 2010 Oct;57(3):206-15. doi: 10.1016/j.neuint.2010.05.011. Epub 2010 May 25.

Abstract

Background: The 1-methyl-4-phenylpyridinium ion (MPP(+)), an inhibitor of mitochondrial complex I, has been widely used as a neurotoxin because it causes a severe Parkinson's disease-like syndrome accompanied by increased levels of intracellular reactive oxygen species (ROS) and apoptotic death. In the present study, we investigated the protective effects of osthole, a coumarin compound extracted from the plant-derived medicine Cnidium monnieri, on MPP(+)-induced cytotoxicity in cultured rat adrenal pheochromocytoma (PC12) cells.

Methods: PC12 cells were treated with MPP(+) 2h after treated with different concentrations of osthole. 24h later, the cell viability, the release of lactate dehydrogenase, the activity of caspase-3 and cytochrome c, the expression ratio of Bax/Bcl-2 and the generation of intracellular ROS were detected.

Results: We found that pretreatment with osthole on PC12 cells significantly reduced the loss of cell viability, the release of lactate dehydrogenase, the activity of caspase-3 and cytochrome c, the increase in Bax/Bcl-2 ratio and the generation of intracellular ROS induced by MPP(+). Moreover, our HPLC analysis of cell extracts confirmed that extracellular osthole does penetrate the cell membrane. Thus osthole may function as an intracellular antioxidant to reduce oxidative stress induced by MPP(+).

Conclusions: Therefore, the present study supports the notion that osthole may be a promising neuroprotective agent for the treatment of neurodegenerative disorders such as Parkinson's disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Caspases / metabolism
  • Cell Nucleus / drug effects
  • Cell Nucleus / ultrastructure
  • Cell Survival / drug effects
  • Chromatography, High Pressure Liquid
  • Coumarins / pharmacology*
  • Cytochromes c / metabolism
  • DNA Fragmentation / drug effects
  • Flow Cytometry
  • L-Lactate Dehydrogenase / metabolism
  • MPTP Poisoning / pathology
  • MPTP Poisoning / prevention & control*
  • Membrane Potentials / drug effects
  • Mitochondria / drug effects*
  • Neuroprotective Agents*
  • PC12 Cells
  • Proto-Oncogene Proteins c-bcl-2 / biosynthesis
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Rats
  • Reactive Oxygen Species / metabolism
  • bcl-2-Associated X Protein / biosynthesis

Substances

  • Coumarins
  • Neuroprotective Agents
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • Reactive Oxygen Species
  • bcl-2-Associated X Protein
  • Cytochromes c
  • L-Lactate Dehydrogenase
  • Caspases
  • osthol