The Effect And Significance Of Calcium Sensing Receptor On The Apoptosis Of Rat Spinal Cord Neurons In Anoxia/Reoxygenation Injury

Objective:To investigate the expression of calcium-sensing receptor(CaSR)in spinal cord neurons and to study the effect and significance of CaSR in spinal cord neurons anoxia/reoxygenation(A/R)injury,in order to point out a new research direction and therapeutic targets.Methods:1.Spinal cord neurons were obtained from the SD rats.Using immunofluorescence technique to identify the spinal cord neurons and observe the expression of CaSR in spinal cord neurons.2.To establish the model of spinal cord neurons anoxia/reoxygenation(A/R)injury.3.The spinal cells were divided into four groups randomly:the control group,anoxia/reoxygenation group(A/R group),A/R+GdCl3 group and A/R+NPS-2390 group.The expression of CaSR in each group was detected by Western-blotting and Fluorescent quantitative PCR.The concentration of intracellular calcium was measured by laser confocal scanning microscope while observing the effection of intracellular calcium by GdCl3 and NPS-2390.4.The apoptotic rate of spinal cells was detected by Tunel assay.Using Western-blotting to detect the expression of Caspase-3,Bax and Bcl-2.Results:1.The expression of CaSR was existed in rat spinal cord neurons and distributed in the cell membrane and cytoplasm.2.The model of spinal cord neurons anoxia/reoxygenation(A/R)injury was established successfully.The expression of CaSR protein in A/R group was higher than in control group.3.Compared to the control group,there was a significant increase in the levels of CaSR protein,CaSR mRNA and the concentration of intracellular calcium in A/Rgroup.However,the above parameters increased more in A/R+GdCl3 group,and decreased in A/R+NPS-2390 group.4.Compared to the control group,there was a significant increase in the levels of Caspase-3,Bax,and the apoptotic rate in A/R group,accompanied with the decreasing of Bcl-2.However,the above parameters increased more in A/R+GdCl3 group,and decreased in A/R+NPS-2390 group.Conclusions:The expression of CaSR was existed in rat spinal cord neurons and distributed in the cell membrane and cytoplasm.During the process of anoxia/reoxygenation injury in rat spinal cord neurons,the expression of calcium sensing receptor and calcium is increased,accompanied with the increasing apoptosis rate.GdCl3 can increase the expression of CaSR and make spinal cord neurons anoxia/reoxygenation injury deeper.However,NPS-2390 has the opposite effect.It is suggested that the intervention of CaSR in spinal cord ischemia-reperfusion injury can be a new way to treat the disease.