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Resveratrol protects primary cortical neuron cultures from transient oxygenglucose deprivation by inhibiting MMP9

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It was recently shown that resveratrol exerts neuroprotective effects against cerebral ischemia in mice. The aim of the present study was to further confirm these effects in in vitro primary cortical neuron cultures with transient oxygenglucose deprivation (OGD), and to investigate whether these effects are due to the inhibition of matrix metalloproteinase9 (MMP9) and of cell apoptosis. Neuronal primary cultures of cerebral cortex were prepared from BALB/c mice embryos (1315 days). Cells from 14- to 16-day cultures were subjected to OGD for 3 h, followed by 21 h of reoxygenation to simulate transient ischemia. Different doses of resveratrol were added into the culture medium during the simulation of transient ischemia. The effect of the extracellular signalregulated kinase (ERK) inhibitor U0126 was studied by adding U0126 (5 µg/µl, 4 µl) into the culture medium during transient ischemia; as a control, we used treatment of cells with 50 µM of resveratrol. Cell viability was investigated using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) reduction assay. Cell apoptosis was assessed by flow cytometry. The effects of resveratrol on the expression of MMP9 were analyzed by western blotting and reverse transcriptionpolymerase chain reaction (RTPCR), while the levels of ERK, phosphorylated (p)ERK, cleaved caspase3, Bax and Bcl2 were measured by western blotting. The results of the MTT assay showed that cell viability is significantly reduced by transient OGD. OGD induced cell apoptosis, the expression of Bax and the activation of caspase3 and ERK, inhibited the expression of Bcl2 and increased the expression of MMP9, while these effects were reversed by treatment with resveratrol. The therapeutic efficacy of resveratrol was shown to be dosedependent, with the most suitable dose range determined at 50100 µM. Treatment with U0126 inhibited MMP9 and Bax expression and caspase3 activation, while it further promoted the expression of the antiapoptotic molecule Bcl2, suggesting that resveratrol inhibits MMP9 expression and cell apoptosis by attenuating the activation of ERK1/2. In conclusion, OGD can induce apoptosis through canonical apoptotic signals and by regulating the expression of MMP9; the antiapoptotic activity of resveratrol and its inhibitory effect on MMP9 expression contribute in the reduced activation of ERK.
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Document Type: Research Article

Affiliations: Department of Neurosurgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China

Publication date: January 1, 2014

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  • Molecular Medicine Reports is a monthly, peer-reviewed journal available in print and online, that includes studies devoted to molecular medicine, underscoring aspects including pharmacology, pathology, genetics, neurosciences, infectious diseases, molecular cardiology and molecular surgery. In vitro and in vivo studies of experimental model systems pertaining to the mechanisms of a variety of diseases offer researchers the necessary tools and knowledge with which to aid the diagnosis and treatment of human diseases.
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