Protective Effect Of Naloxone On The Damage Of Cultured Rat Cortical Neurons

Objective:To investigate the damage of anoxic and LPS(lipopolysaccharide) condition on thecultured rat cortical neurons and the protective effects of naloxone.Methods:1. Cortical neurons cultured for 12d were randomly divided into three groups of control, anoxic and anoxic plus naloxone treated. They were exposed to anoxic environment for 6h and then cultured for 24h under normoxic condition.2. Primitive rat cortical neurons were randomly divided into three groups: control group, LPS group, naloxone plus LPS group.3. Observe the variation of survival rate and morphology of neurons under different treating conditions, measure the content of lactate dehydrogenase (LDH) in the solution and test the apoptosis of neurons by the method of TUNEL. Using flow cytometry, measure the apoptotic rate of neurons and the variation of intracellular calcium and mitochondrial membrane potential.Results:1. After enzyme digestion for cortical neurons of newborn rats, above 90% of primitive cells were neurons.2. The pathological process of neuron damage caused by 6h anoxia and 24h normoxia culture was similar to that of clinical neurons damage caused by ischemia, anoxia and reperfusion injury.3. Anoxia and lipopolysaccharide treatment can lead to pathological damage of culturedrat cortical neurons. Those pathologic changes included intracellular calcium overload,abnormal, increased permeability of cellular membrane and increase in neuron apoptosisand death.4. Naloxone can ameliorate the toxicity damage of neurons caused by anoxia and LPS,behaving as correcting intracellular calcium overload, improving energy metabolism,maintaining permeability of cellular membrane and reducing neuron apoptosis and death.Conclusion:Anoxia and LPS can lead to severe damages of cultured cortical neurons, which include intracellular calcium overload, abnormal energy metabolism, increased permeability of cellular membrane and increase in neuron apoptosis and death. Naloxone can ameliorate the damage of neurons caused by anoxia and LPS, reduce apoptosis of neurons and suppress the increasing of the intracellular calcium, which suggest that naloxone can protect rat cortical neurons from damage under anoxia and LPS condition.