Effects of hydrogen sulfide on mouse cortical neuronal injuries induced by oxygen gloucose deprivation/reoxygenation in vitro

doi:10.3969/j.issn.0529-1356.2014.03.004

Abstract

Abstract:

Objective To explore the effects of H₂S on neuronal injuries induced by oxygen glucose deprivation/reoxygenation (OGD/R) in cortical neurons. Methods For OGD, the primary cultured cortical neurons were incubated with glucose-free EBSS media for 4h in N₂/CO₂/O₂ (93%/5%/2%) atmosphere. Thereafter, the media were replaced by Neurobasal/B27 culture media and the neurons were incubated for 12 h in a 5% CO₂ incubator at 37℃. NaHS was used as a H₂S donor and cell survival rate was determined by cell counting kit 8(CCk-8). ［Ca ²⁺ ]i was determined using fura-2/AM and fluorescence microscopic imaging systems. The release rate of lactate dehydrogenase (LDH) was determined by lactate dehydrogenase assay kit, and cell damage was analyzed by staining of propidium iodide (PI). Results After pretreated with 200, 300 and 600 μmol/L sodium hydrosulfide (NaHS) for 30min before OGD/R, the cell survival rate of neurons significantly increased (n=4). ［Ca ²⁺］I(n=5), LDH release rate (n=4) and cell damage percentage (n=6) in the neuron pretreated with 300 μM NaHS were significantly lower than those in ODG/R cells. Treatment with 10μmol/L calcium chelator BAPTA also reduced the LDH release rate and cell damage percentage induced by ODG/R in neurons. Conclusion The results indicate that H₂S may inhibit the OGD/R induced damage in cortical neurons via reducing calcium overload of neurons.

Key words: Oxygen-glucose deprivation/reoxygenation, Hydrogen sulfide, Cell injury, Calcium overload, Fluorescence microscopic imaging, Mouse

WU Ji-yun WEI Ci-zhao XU Yue-qing LIU Lu-kuan ZHANG Yang-ping WEI Chu-rong MAO Mu-hua LUO You-gen*. Effects of hydrogen sulfide on mouse cortical neuronal injuries induced by oxygen gloucose deprivation/reoxygenation in vitro[J]. AAS, 2014, 45(3): 316-320.

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