大鼠脊髓神经元缺氧复氧损伤中钙敏感受体对其凋亡的影响及意义  

Effect of Calcium-sensing Receptor on the Apoptosis of Rat Spinal Cord Neurons in Anoxia/Reoxygenation Injury and Its Significance

作　　者：何炜[1] 孙继芾[1] 蒋润宇 张志坚[2] 陈谦 黄永辉[1] 

机构地区：[1]江苏大学附属医院骨科,江苏镇江212001 [2]江苏大学医学院基础与临床医学研究中心,江苏镇江212013

出　　处：《中国医学科学院学报》2017年第5期625-630,共6页Acta Academiae Medicinae Sinicae

摘　　要：目的探究大鼠脊髓神经元缺氧复氧损伤中钙敏感受体对其凋亡的影响及意义。方法将新生SD大鼠脊髓神经元随机分为正常对照组、缺氧复氧组、激动剂(Gd Cl3)组和抑制剂(NPS-2390)组4组。采用95%氮气通入置有细胞培养板的密闭通气盒中15 min,37℃培养箱内培养1 h,再用含2%B27、10%胎牛血清的Neurobasol-A液培养细胞24 h复制缺氧复氧模型。应用免疫荧光技术鉴定脊髓神经元并观察钙敏感受体的表达定位,Western blot法检测各组钙敏感受体及凋亡相关蛋白Caspase-3、Bax、Bcl-2的表达水平,激光共聚焦显微镜测定细胞内游离钙的变化,Tunel法观察各组细胞凋亡情况。结果缺氧复氧组的钙敏感受体(t=5.462,P=0.006)、细胞内游离钙(t=8.573,P=0.001)、细胞凋亡(t=4.899,P=0.008)、Caspase-3(t=5.118,P=0.007)和Bax(t=10.930,P=0.001)表达水平均明显高于对照组,激动剂组的钙敏感受体(t=4.975,P=0.008)、细胞内游离钙(t=4.899,P=0.008)、细胞凋亡(t=7.746,P=0.002)、Caspase-3(t=4.776,P=0.009)和Bax(t=5.281,P=0.006)表达水平均明显高于缺氧复氧组,抑制剂组的钙敏感受体(t=3.674,P=0.021)、细胞内游离钙(t=3.846,P=0.018)、细胞凋亡(t=4.281,P=0.013)、Caspase-3(t=3.521,P=0.024)和Bax(t=3.473,P=0.026)表达水平均明显低于缺氧复氧组;缺氧复氧组的Bcl-2表达水平明显低于对照组(t=6.242,P=0.003),激动剂组明显低于缺氧复氧组(t=3.028,P=0.004),抑制剂组明显高于缺氧复氧组(t=2.840,P=0.047)。结论在缺氧复氧损伤过程中,钙敏感受体表达增多,细胞内游离钙增加,脊髓神经元凋亡增多。Objective To investigate the effect and significance of calcium-sensing receptor( Ca SR)on the apoptosis of rat spinal cord neurons in anoxia/reoxygenation( A/R) injury. Methods The spinal cells were in ischemia and hypoxia environment for 1 h and in normal environment for 24 h to establish a model of A/R. After spinal A/R model was established,the spinal cells were divided into four groups randomly: the control group,A/R group,A/R + Gd Cl3 group,and A/R + NPS-2390 group. The expression of Ca SR in each group was detected by immunofluorescence and Western blotting. The concentration of intracellular calcium was measured by laser confocal scanning microscopy. The expressions of Caspase-3,Bax,and Bcl-2 were detected by using Western blotting. The apoptotic rate of spinal cells was detected by Tunel assay. Results Compared to thecontrol group,there was a significant increase in the level of Ca SR( t = 5. 462,P = 0. 006),the concentration of intracellular calcium( t = 8. 573,P = 0. 001),the apoptotic rate( t = 4. 899,P = 0. 008),Caspase-3( t =5. 118,P = 0. 007),and Bax( t = 10. 930,P = 0. 001) in A/R group. Compared to the A/R group,there was a significant increase in the level of Ca SR( t = 4. 975,P = 0. 008),the concentration of intracellular calcium( t = 4. 899,P = 0. 008),the apoptotic rate( t = 7. 746,P = 0. 002),Caspase-3( t = 4. 776,P = 0. 009),and Bax( t = 5. 281,P = 0. 006) in A/R + Gd Cl3 group. Compared to the A/R group,there was a significant decrease in the level of Ca SR( t = 3. 674,P = 0. 021),the concentration of intracellular calcium( t = 3. 846,P =0. 018),the apoptotic rate( t = 4. 281, P = 0. 013),Caspase-3( t = 3. 521,P = 0. 024),and Bax( t =3. 473,P = 0. 026) in A/R + NPS-2390 group. However,compared to the control group,there was a significant decrease in the level of Bcl-2( t = 6. 242,P = 0. 003) in A/R group. Compared to the A/R group,there was a significant decrease in the level of Bcl-2( t = 3. 028,P = 0. 004) in A/R + Gd Cl3 group. Compared to the A/R group,there was a significant increase

关 键 词：缺氧复氧 钙敏感受体 凋亡 脊髓神经元 

分 类 号：R364.1[医药卫生—病理学]