Protective Effects of Berberine on Oxygen-Glucose Deprivation/Reperfusion on Oligodendrocyte Cell Line (OLN-93)

Shabnam Nadjafi, Soltan-Ahmad Ebrahimi, Nahid Rahbar-Roshandel

Abstract


Background: Oligodendrocytes, the myelinating glial cells of central nervous system, are highly vulnerable to ischemic‑induced excitotoxic insult, a phenomenon in which calcium overload triggers cell death. Berberine is an alkaloid extracted from medicinal herbs as Coptidis Rhizoma with several pharmacological effects like inhibition of neuronal apoptosis in cerebral ischemia.

Methods: We examined the effects of berberine (0.5-4 μM) and glutamate receptors antagonists (MK‑801 [10 μM] and NBQX [30 μM]) on OLN‑93 cell line (a permanent immature rat oligodendrocyte) during (30, 60, 240 min) oxygen‑glucose deprivation (OGD)/24 h reperfusion. The cells were cultured in 12-well plates. The cells were exposed to glucose‑free medium and hypoxia in a small anaerobic chamber. Cell viability was evaluated by MTT (3-[4,5‑Dimethylthiazol‑2‑yl]-2,5‑diphenyl‑tetrazolium bromide) assay. The intracellular calcium levels also were evaluated by Ca2+‑sensitive indicator Fura-2/AM in presence or absence of berberine (2 μM) during 30 min chemical OGD by NaN3 (20 mM). Student’s t‑test and ANOVA were used for statistical analysis.

Results: Berberine, MK‑801and NBQX significantly increased oligodendrocyte viability in all 3 time‑scheduled oxygen‑glucose deprivation/reperfusion. Berberine at 2 μM produced peak of protection, and increased cell viability to 83%, 77%, and 79% during 30, 60, 240 min ischemic experiments, respectively (P < 0.001). Berberine significantly attenuated intracellular Ca2+ rise induced by chemical ischemia, and this effect of berberine was significantly stronger than MK‑801 and NBQX (P < 0.001).

Conclusions: We concluded that berberine protected OLN‑93 oligodendrocyte against ischemic induced excitotoxic injury. Attenuation of intracellular Ca2+ overload by berberine may be the key mechanism that saved OLN‑93 from excitotoxicity damage.

Keywords: Berberine, calcium, excitotoxicity, OLN-93 cell line, oxygen‑glucose deprivation

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