脑缺血后处理对大鼠大脑海马区Notch1、Cyclin D1和CDK4蛋白表达的影响

doi:10.13481/j.1671-587x.20170614

Abstract

Abstract: Objective:To investigate the effects of cerebral ischemic postconditioning (CIP) on the expressions of Cyclin D1 and CDK4 proteins in hippocampal neurons of the rats with global cerebral ischemia and reperfusion (I/R) based on Notch1 signaling pathway,and to explore the mechanisms. Methods:A total of 128 healthy male SD rats were randomly divided into sham operation group, I/R group (modified Pulsinelli four vessel occlusion method), CIP group(repeated 3 times of reperfusion or blocking blood vessel before complete reperfusion) and DAPT group(intraperitoneal injection of 10 mg·kg^-1·d^-1 DAPT 3 h before CIP),and there were 32 rats in each group. HE staining was used to observe the morphology of the neurons at 6, 24, 48 and 72 h after ischemia;immunohistochemical staining was used to observe the expressions of Cyclin D1, CDK4 and Notch1 in the hippocampus of the rats;Western blotting method was used to observe the expression levels of Cyclin D1, CDK4 and Notch1 proteins in the hippocampus of the rats at different time points. Results:The HE staining results showed that compared with sham operation group, the structure of neurons in hippocampus area of the rats in I/R group was damaged and the survival rate of neurons was significantly decreased(P<0.05).Compared with I/R group, the survival rates of neurons in the hippocampus area of the rats in CIP group were significantly increased at the corresponding time(P<0.05).Compared with CIP group, the survival rates of neurons in hippocampus area of the rats in DAPT group at the corresponding time were significantly decreased (P<0.05).The immunohistochemical staining results showed that compared with sham operation group,the number of cells with positive Notch1, Cyclin D1 and CDK4 in I/R group was increased (P<0.05).Compared with I/R group, the number of Cyclin D1 and CDK4 positive cells in CIP group was decreased (P<0.05), and the number of Notch1 positive cells of was significantly increased (P<0.05).Compared with CIP group, the number of Cyclin D1 and CDK4 positive cells in DAPT group was increased (P<0.05), and the number of Notch1 positive cells was significantly decreased(P<0.05).The Western blotting results showed that compared with sham operation group, the expression levels of Notch1, Cyclin D1 and CDK4 proteins in the hippocampus of the rats in I/R group were increased (P<0.05);compared with I/R group, the expression levels of Cyclin D1 and CDK4 proteins in the hippocampus of the rats in CIP group were significantly decreased (P<0.05), and the expression level of Notch1 protein in the hippocanpus of the rats was significantly increased (P<0.05);compared with CIP group, the expression levels of Cyclin D1 and CDK4 proteins in the hippocampus of the rats in DAPT group were increased (P<0.05), and the expression level of Notch1 protein was significantly decreased (P<0.05). Conclusion:CIP may play an important role in protecting the neurons by increasing the activity of Notch1 and inhibiting the expressions of Cyclin D1 and CDK4.

Key words: ischemic postconditioning, Notch1, nerve protection, Cyclin D1, CDK4

CLC Number:

R473.74

LIU Yao, ZHAO Xiaoyun, ZHAO Yaning, LI Jianmin, LI Jianing. Effects of cerebral ischemic postconditioning on expressions of Notch1, Cyclin D1 and CDK4 proteins in hippocampus of rats[J].Journal of Jilin University Medicine Edition, 2017, 43(06): 1142-1147.

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Effects of cerebral ischemic postconditioning on expressions of Notch1, Cyclin D1 and CDK4 proteins in hippocampus of rats

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