| Objective: To investigate the expression and role of calcium sensing receptor(CaSR) and calpain in apoptosis of rat spinal cord neurons induced by anoxia/reoxygenation(A/R). Methods:After spinal anoxia/reoxygenation model was established by culturing primary spinal cell of neonatal rat in vivo.The spinal cells were divided into four groups randomly:the control group,A/R group,calcimimetic group(GdCl3 group),inhibitor group.The apoptotic rate of spinal cells was determined by Hoechest33258 assay, the expression of Ca SR and Calpain was detected by fluorescence immunoassay and western blotting. Results:1. Calcium sensing receptor and calpain are distributed in cell membrane, cytoplasm and neurite of the Spinal cord neurons.2.compared with control group, expression of CaSR, calpain and Bax protein rised in hypoxia / reoxygenation group,and the apoptosis rate was increased too.3.Compared with hypoxia/reoxygenation group,the expression of CaSR, calpain and Bax protein in agonist group increased, while the inhibitor group had opposite effect.Conclusion:1.Calcium sensing receptor and calpain eexists in the rat spinal cord neurons, which are distributed in spinal cord neurons in cell membrane, cytoplasm and protrusion.2.Hypoxia can activate Ca SR, and can increase apoptosis. The use of agonist GdCl3 can aggravate the original hypoxia and reoxygenation injury,increase apoptosis; inhibitor’s effect with the contrary effect. It showed that Ca SR participated in the process of apoptosis.3.CaSR excitement or inhibition, are accompanied by the expression of calpain increased or decreased, indicating that in the process of apoptosis and Ca SR and calpain exists positive correlation.4.Inhibitor NPS2390 can significantly inhibit cell apoptosis, reduce cell hypoxia and reoxygenation injury. Therefore, the specificity of CaSR inhibitors probably will become the treatment of spinal cord ischemia-reperfusion injury in a effective way. |
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