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Planta Med 2003; 69(6): 506-512
DOI: 10.1055/s-2003-40647
Original Paper
Pharmacology
© Georg Thieme Verlag Stuttgart · New York

Protective Effects of Fangchinoline and Tetrandrine on Hydrogen Peroxide-Induced Oxidative Neuronal Cell Damage in Cultured Rat Cerebellar Granule Cells

Sang Bum Koh1 , Ju Yeon Ban1 , Bo Young Lee1 , Yeon Hee Seong1
  • 1College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, Korea
Weitere Informationen

Publikationsverlauf

Received: August 13, 2002

Accepted: March 8, 2003

Publikationsdatum:
16. Juli 2003 (online)

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Abstract

The present study was performed to examine the neuroprotective effects of fangchinoline (FAN) and tetrandrine (TET), bis-benzylisoquinoline alkaloids, which exhibit the characteristics of Ca2+ channel blockers, on H2O2-induced neurotoxicity using cultured rat cerebellar granule neurons. H2O2 produced a concentration-dependent reduction of cell viability, which was blocked by (5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801), an N-methyl-D-aspartate (NMDA) receptor antagonist, verapamil, an L-type Ca2+ channel blocker, and N G-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor. Pretreatment with FAN and TET over a concentration range of 0.1 to 10 μM significantly decreased the H2O2-induced neuronal cell death as assessed by a trypan blue exclusion test, a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and the number of apoptotic nuclei. In addition, FAN and TET inhibited the H2O2-induced elevation of glutamate release into the medium, elevation of the cytosolic free Ca2+ concentration ([Ca2+]c), and generation of reactive oxygen species (ROS). These results suggest that FAN and TET may mitigate the harmful effects of H2O2-induced neuronal cell death by interfering with the increase of [Ca2+]c, and then by inhibiting glutamate release and generation of ROS.

Abbreviations

AP5:D(-)-2-amino-5-phosphonopentanoic acid

DMSO:dimethyl sulfoxide

FAN:fangchinoline

H2DCF-DA:2′,7′-dichlorodihydrofluorescin diacetate

MK-801:(5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,20-imine

MTT:3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide

L-NAME:N G-Nitro-L-arginine methyl ester

NMDA:N-methyl-D-aspartate

TET:tetrandrine

Key words

Fangchinoline - tetrandrine - H2O2 - neurotoxicity - cytosolic free Ca2+ concentration - cerebellar granule cells

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Prof. Dr. Yeon Hee Seong

College of Veterinary Medicine and Research Institute of Veterinary Medicine

Chungbuk National University

San 48

Kaeshin-Dong

Heungduk-Ku

Cheongju

Chungbuk 361-763

Republic of Korea

eMail: vepharm@chungbuk.ac.kr

Fax: +82-43-267-3150