Neuroprotective Effects Of Hypoxia Preconditioning And 3-n-butylphalide In Primary Culture Rat Hippocampus Neurons

Objective: (1)To observe neuroprotective effects of hypoxia preconditioning(HP) in primary culture SD rat hippocampus neurons and investigate its possible mechanisms;(2)To observe neuroprotective effects of 3-n-butylphalide(NBP) against oxygen glucose deprivation /reoxygenation (OGD / R) injury in primary culture SD rat hippocampus neurons and investigate its possible mechanisms to against ischemic reperfusion injury.Methods :After 7-day primary cultured hippocampal neurons of newborn SD rats(< 24h)in vitro,(1)established the HP model , hippocampal neurons were divided into three groups: normal control group, pure hypoxia group and HP group.⑵Established OGD30min/R8h model, hippocampal neurons were divided into five groups: normal control group, OGD/R model group, OGD/R+NBP group(0.1μmol/L,1μmol/L and 10μmol/L group). Apoptotic cells were analyzed by Hoechest33258 staining. Bcl-2 and NGB expression of cells in each group were analyzed by immunocytochemistry. The mRNA expression of portions Bcl-2 and NGB in each group were analyzed by RT-PCR.Results⑴Comparing with pure hypoxia group, HP group showed a significantly decreased neuronal apoptosis and increased expression of proteins Bcl-2 and NGB,and their mRNA (P < 0.01);⑵Comparing with OGD/R model group,NBP group showed a significantly decreased neuronal apoptosis and increased expression of proteins Bcl-2 and NGB (P < 0.05);⑶Comparing between each different concentration of NBP drugs showed a significantly decreased neuronal apoptosis and increased expression of proteins Bcl-2 and NGB with the increasing of NBP concentration(P < 0.01) .Conclusion (1)HP inhibit apoptosis of hippocampal neurons with hypoxia injury and play endogenous neuroprotection by up regulation the mRNA of Bcl-2 and NGB, and increasing the expression of protein Bcl-2 and NGB;⑵NBP inhibit apoptosis of hippocampal neurons with OGD/R injury and play exogenous neuroprotection by increasing of proteins Bcl-2 and NGB; The exogenous neuroprotection is enhanced with the increasing of NBP concentration.⑶The neuroprotection of HP and NBP may be have same possible mechanisms of increasing the expression of Bcl-2 and NGB proteins.