| 分泌型糖蛋白Reelin对N-甲基-D-天冬氨酸（NMDA）诱导的视网膜神经节细胞凋亡的抑制作用 |
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| 引用本文： | 王艳婷,金学民,李晓华,赵朝霞.分泌型糖蛋白Reelin对N-甲基-D-天冬氨酸（NMDA）诱导的视网膜神经节细胞凋亡的抑制作用[J].眼科新进展,2021,0(6):518-521. |
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| 作者姓名： | 王艳婷 金学民 李晓华 赵朝霞 |
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| 作者单位： | 450000 河南省郑州市，河南省人民医院眼科(王艳婷，李晓华，赵朝霞)；450000 河南省郑州市，郑州大学第一附属医院眼科（金学民） |
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| 摘 要： | 目的 探讨分泌型糖蛋白Reelin对N-甲基-D-天冬氨酸（NMDA）诱导的视网膜神经节细胞（RGC）凋亡的抑制作用。方法 将大鼠RGC-5分为对照组、NMDA组、Reelin类似物组、Reelin阴性对照组和Reelin抑制物组。对照组细胞用含双抗的氨基酸双抗培养基培养，NMDA组在对照组细胞培养基中添加100 μmol·L-1 NMDA后进行细胞培养，Reelin类似物组、Reelin阴性对照组和Reelin抑制物组在NMDA组细胞培养基的基础上利用Lipofectamine 2000将Reelin类似物、阴性对照和抑制物转染RGC-5，共同孵育24 h。用Annexin V FITC/碘化丙啶联合流式细胞仪检测细胞凋亡率，Western blot检测细胞内Reelin、磷酸化-Tau蛋白（p-Tau）、糖原合成酶激酶-3β (GSK-3β)和细胞周期素依赖蛋白激酶5（CDK5）表达情况。结果 对照组RGC-5细胞凋亡率为（9.30±0.74）%，NMDA组为（25.40±1.13）%，NMDA组高于对照组，差异有统计学意义（t=6.311,P<0.001）。NMDA组RGC-5内Reelin蛋白表达水平低于对照组，p-Tau、GSK-3β和CDK5蛋白表达水平均高于对照组，差异均有统计学意义(均为P<0.001)。Reelin类似物组细胞凋亡率为（9.23±0.24）%，低于Reelin阴性对照组［（14.33±1.02）%］和Reelin抑制物组［（26.81±1.15）%］，各组细胞凋亡率差异均有统计学意义（均为P<0.05）。Reelin类似物组RGC-5内p-Tau、GSK-3β蛋白表达水平均低于Reelin阴性对照组和Reelin抑制物组，Reelin抑制物组RGC-5内p-Tau、GSK-3β蛋白表达水平均高于Reelin阴性对照组，差异均有统计学意义（均为P<0.05）；CDK5蛋白表达水平在3组中基本一致，差异无统计学意义（P>0.05）。结论 Reelin对NMDA 诱导损伤的RGC具有保护作用，这种作用是通过下调GSK-3β表达抑制Tau蛋白磷酸化，进一步抑制细胞凋亡而实现的。
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| 关 键 词： | Reelin 青光眼 视网膜神经节细胞 Tau蛋白 细胞凋亡 |
Secreted glycoprotein Reelin prevents N-methyl-D-aspartate-induced apoptosis of retinal ganglion cells |
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| WANG Yanting,JIN Xuemin,LI Xiaohua,ZHAO Zhaoxia.Secreted glycoprotein Reelin prevents N-methyl-D-aspartate-induced apoptosis of retinal ganglion cells[J].Recent Advances in Ophthalmology,2021,0(6):518-521. |
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| Authors: | WANG Yanting JIN Xuemin LI Xiaohua ZHAO Zhaoxia |
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| Affiliation: | 1.Department of Ophthalmology of Henan Provincial People’s Hospital，Zhengzhou 450000，Henan Province,China2.Department of Ophthalmology，the First Affiliated Hospital of Zhengzhou University，Zhengzhou 450000，Henan Province,China |
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| Abstract: | Objective To investigate the inhibitory effect of secreted glycoprotein Reelin on N-methyl-D-aspartate (NMDA)-induced apoptosis of retinal ganglion cells (RGC).Methods RGC-5 of rats were divided into control group, NMDA group, Reelin analog group, Reelin negative control group and Reelin inhibitor group. Cells in the control group were cultured with double-antibody-containing amino acid double-antibody (DEME) medium, the NMDA group was cultured with DEME medium plus 100 μmol·L-1 NMDA. Reelin analog group, Reelin negative control group and Reelin inhibitor group were cultured with DEME medium plus 100 μmol·L-1 NMDA, and then RGC-5 cells were transfected with the Reelin analog, negative control and inhibitor using Lipofectamine 2000, respectively, for 24 hours . The apoptosis rates were detected by Annexin V FITC/propidium iodide combined with flow cytometry, while Western blot was used to detect the expression levels of Reelin, phosphorylated-Tau protein (p-Tau), Glycogen synthasc kinase-3β (GSK-3β), cyclin-dependent kinase 5 (CDK5) protein.Results The apoptosis rate of RGC-5 cells in the control group was (9.30±0.74)%, the NMDA group was (25.40±1.13)%, and the apoptosis rate of NMDA group was higher than that of the control group, and the difference was statistically significant (t=6.311, P<0.001). The expression of Reelin protein in RGC-5 in the NMDA group was lower than that in the control group, and the expression levels of p-Tau, GSK-3β and CDK5 protein were higher than those in the control group, and the differences were statistically significant (all P<0.001). The apoptosis rate of the Reelin analog group was (9.23±0.24) %, which was lower than that of the Reelin negative control group ［(14.33±1.02)%］ and the Reelin inhibitor group ［(26.81±1.15)%］, and the differences were statistically significant (both P<0.05). The expression levels of p-Tau and GSK-3β of RGC-5 in the Reelin analog group were lower than those in the Reelin negative control group and Reelin inhibitor group. The expression levels of p-Tau and GSK-3β in RGC-5 in the Reelin inhibitor group were higher than those in the Reelin negative control group, and the differences were statistically significant (all P<0.05), but CDK5 protein expression level was basically the same in the three groups, and the difference was not statistically significant (P>0.05).Conclusion Reelin has a protective effect against NMDA-induced damage to RGC. This effect is achieved through Reelin down regulating the expression of GSK-3β, thereby inhibiting Tau protein phosphorylation and further inhibiting cell apoptosis. |
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| Keywords: | Reelin glaucoma retinal ganglion cells Tau protein cell apoptosis |
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