title: Berberine exerts neuroprotective actions against in vitro ischemia-induced neuronal cell damage in organotypic hippocampal slice cultures: involvement of B-cell lymphoma 2 phosphorylation suppression.
abstract: In this study we elucidated the effects of berberine, a major alkaloid component contained in medicinal herbs, such as Phellodendri Cortex and Coptidis Rhizoma, on ischemic neuronal damage in mouse organotypic hippocampal slice cultures (OHSCs) caused by oxygen and glucose deprivation (OGD) and N-methyl-D-aspartate (NMDA) -type glutamate receptor stimulation. Hippocampal slices obtained from 7-d-old ICR mice were cultured for 10 d before the experiments. Ischemia-related damage was induced by OGD (5, 15, 45 min) or NMDA (10 microM) treatment, and was evaluated by measuring propidium iodide (PI) uptake. Levels of apoptotic marker proteins, B-cell lymphoma 2 (Bcl-2) and phosphorylated-Bcl-2 (p-Bcl-2), in the OHSCs were measured as indices of biochemical neuronal cell damage by Western blotting. Berberine (5, 25 microM) or the NMDA antagonist MK-801 (25 microM) was added to the medium 30 min before OGD or NMDA treatment. OGD time-dependently increased PI uptake of the OHSCs. Both berberine (5, 25 microM) and MK-801 (25 microM) significantly inhibited PI uptake at 24 h after 45-min OGD treatment and PI uptake in OHSCs exposed to NMDA for 24 h. OGD treatment also significantly increased the level of p-Bcl-2 but not that of Bcl-2 or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in OHSCs. Berberine (5-25 microM) significantly suppressed the OGD-induced increase of p-Bcl-2 level in OHSCs when tissue was exposed to the alkaloid prior to OGD or simultaneously with OGD. These findings suggest that berberine has protective effects against ischemic damage in mouse OHSCs and that the effects are at least partly mediated by suppression of Bcl-2 phosphorylation.
authors.affiliation: Division of Medicinal Pharmacology, Institute of Natural Medicine, University of Toyama, Japan.
authors.forename: Hu-Shan
authors.initials: HS
authors.lastname: Cui
authors.forename: Kinzo
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Yukihisa
authors.initials: Y
authors.lastname: Murakami
authors.forename: Hitomi
authors.initials: H
authors.lastname: Hori
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhao
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Obi
mesh_terms: D000818:Animals; D001599:Berberine / Q000737:chemistry* / Q000494:pharmacology* / Q000627:therapeutic use; D016923:Cell Death / Q000187:drug effects; D004281:Docosahexaenoic Acids / Q000494:pharmacology* / Q000627:therapeutic use; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D005947:Glucose / Q000172:deficiency; D006624:Hippocampus / Q000187:drug effects / Q000473:pathology* / Q000503:physiopathology; D000860:Hypoxia / Q000150:complications / Q000188:drug therapy; D007511:Ischemia / Q000150:complications / Q000473:pathology; D007555:Isoxazoles / Q000494:pharmacology / Q000627:therapeutic use; D051379:Mice; D008813:Mice, Inbred ICR; D016202:N-Methylaspartate / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects*; D009924:Organ Culture Techniques; D010766:Phosphorylation; D011419:Propidium; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology / Q000627:therapeutic use; D013997:Time Factors; D000096926:Benzenesulfonamides
chemical_list: D001599:Berberine; D004281:Docosahexaenoic Acids; D018690:Excitatory Amino Acid Agonists; D005947:Glucose; D007555:Isoxazoles; D016202:N-Methylaspartate; D011419:Propidium; D019253:Proto-Oncogene Proteins c-bcl-2; D013449:Sulfonamides; C497709:N-(2-(4-methoxybenzenesulfonamide)phenyl)-3,5-dimethyl-4-isoxazolcarboxamide; D000096926:Benzenesulfonamides