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

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (06): 1142-1147.doi: 10.13481/j.1671-587x.20170614

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

刘瑶¹, 赵晓云¹, 赵雅宁¹, 李建民², 李佳宁³

1. 华北理工大学护理与康复学院, 河北唐山 063000;
2. 华北理工大学附属医院神经外科, 河北唐山 063000;
3. 河北省唐山市人民医院急诊科, 河北唐山 063000

收稿日期:2016-11-20 出版日期:2017-11-28 发布日期:2017-12-01
通讯作者: 赵雅宁,教授,硕士研究生导师(Tel:0315-4839888,E-mail:zyning789@126.com) E-mail:zyning789@126.com
作者简介:刘瑶(1992-),女,河北省唐山市人,在读医学硕士,主要从事慢性病方面的研究。
基金资助:
河北省卫计委科研重点项目资助课题（ZD2010106）

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

LIU Yao¹, ZHAO Xiaoyun¹, ZHAO Yaning¹, LI Jianmin², LI Jianing³

1. College of Nursing and Rehabilitation Medicine, North China University of Science and Technology, Tangshan 063000, China;
2. Department of Neurosurgery, Affiliated Hospital, North China University of Science and Technology, Tangshan 063000, China;
3. Department of Emergency, Tangshan People's Hospital, Hebei Province, Tangshan 063000, China

Received:2016-11-20 Online:2017-11-28 Published:2017-12-01

摘要/Abstract

摘要： 目的：探讨基于Notch1信号通路观察缺血后处理（CIP）对全脑缺血再灌注（I/R）大鼠海马区神经元中Cyclin D1和CDK4蛋白表达的影响，阐明其机制。方法：128只健康的雄性SD大鼠随机分为假手术组、I/R组（改良的Pulsinelli四血管阻断法制作）、CIP组（在彻底再灌注之前进行反复3次的再通/阻断血管）和DAPT组（实施CIP之前3 h按10mg·kg^-1·d^-1腹腔注射DAPT），每组32只。分别在缺血后6、24、48和72 h采用HE染色观察各组大鼠海马区神经元表现并计算存活率，免疫组织化学染色观察CyclinD1、CDK4和Notch1表达的细胞，Western blotting法观察各时间点大鼠海马区中Cyclin D1、CDK4和Notch1蛋白的表达水平。结果：HE染色，与假手术组比较，I/R组大鼠海马区神经元结构损伤，各时间点海马区神经元存活率明显降低（P<0.05）；与I/R组比较，CIP组相应时间点大鼠海马区神经元存活率明显升高（P<0.05）；与CIP组比较，DAPT组相应时间点大鼠海马区神经元存活率明显降低（P<0.05）。免疫组织化学染色，与假手术组比较，I/R组大鼠海马区Notch1、Cyclin D1和CDK4阳性细胞数增加（P<0.05）；与I/R组比较，CIP组大鼠海马区Cyclin D1和CDK4阳性细胞数增加（P<0.05），Notch1阳性细胞数明显增加（P<0.05）；与CIP组比较，DAPT组大鼠海马区Cyclin D1和CDK4阳性细胞数增加（P<0.05），Notch1阳性细胞数明显减少（P<0.05）。Western blotting法，与假手术组比较，I/R组大鼠海马区Notch1、Cyclin D1和CDK4蛋白表达水平升高（P<0.05）；与I/R组比较，CIP组大鼠海马区Cyclin D1和CDK4蛋白表达水平明显降低（P<0.05），Notch1蛋白表达水平明显升高（P<0.05）；与CIP组比较，DAPT组的Cyclin D1和CDK4蛋白表达水平明显升高（P<0.05），Notch1蛋白表达水平明显降低（P<0.05）。结论：CIP对神经元的保护作用可能是通过提高Notch1蛋白活性和抑制Cyclin D1和CDK4蛋白实现的。

关键词: 神经保护, Cyclin D1, CDK4, 缺血后处理, Notch1

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

中图分类号:

R473.74

刘瑶, 赵晓云, 赵雅宁, 李建民, 李佳宁. 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[J]. 吉林大学学报(医学版), 2017, 43(06): 1142-1147.

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.

参考文献

[1] Zhao ZQ, Corvera JS, Halkos ME, et al. Inhibition of myocardial injury by ischemic postconditioning during reperfusion:comparison with ischemic preconditioning[J]. Am J Physiol Heart Circ Physiol, 2003, 285(2):579-588.
[2] 刘瑶,赵雅宁,李建民,等. P38MAPK通路对脑缺血后处理大鼠海马自噬的影响[J]. 西安交通大学学报:医学版,2017,38(4):522-528.
[3] SunF, MaoX, XieL, et al. Notch1 signaling modulates neuronal progenitor activity in the subventricular zone in response to aging and focal ischemia[J]. Aging Cell, 2013, 12(6):978-987.
[4] 金文虎,王达利,聂开瑜,等. 增生性瘢痕成纤维细胞中Cyclin D1,CDK2,CDK4作用及与细胞周期的相关性[J]. 中国组织工程研究,2012,16(24):4422-4426.
[5] Zhao Y, Chen X, Ma L, et al. Electroacupuncture pretreatment induces tolerance against focal cerebral ischemia through activation of canonical Notch pathway[J]. BMC Neurosci, 2012, 13(1):111.
[6] 李兵,章翔,蒋晓帆,等. 改良四血管阻塞法建立大鼠全脑缺血模型[J].中华神经外科疾病研究杂志,2005,4(2):110-113.
[7] 彭荣琳,涂荣会,郑国军.STEMI急诊介入治疗中缺血后处理对心肌的保护作用[J].心血管康复医学杂志,2015,24(3):277-279.
[8] 吉斐,华赟鹏,简佩恩,等. 缺血后处理对肝硬化大鼠肝脏缺血-再灌注损伤保护作用的免疫机制[J]. 中华肝脏外科手术学电子杂志,2014,3(6):371-374.
[9] 王磊,刘修恒,陈晖.肾脏缺血后处理对缺血再灌注损伤保护作用的研究进展[J].职业与健康,2015,31(16):2292-2294,2298.
[10] 张浩,伍火志,马明生.缺血后处理对肺移植早期NF-κB激活信号通路影响的研究[J]. 中国现代医学杂志,2015,25(7):12-17.
[11] 韩冬,孙淼,何平平,等. 缺血后处理对大鼠脑缺血再灌注后紧密连接的保护作用及ZO-1蛋白表达的影响[J]. 中风与神经疾病杂志,2015,32(4):296-298.
[12] 李鑫,吕达平,姚明,等. 缺血后处理通过NMDA受体对全脑缺血大鼠的神经保护作用[J]. 中风与神经疾病杂志,2015,32(10):910-913.
[13] Wu X, Fleming A, Ricketts T, et al. Autophagy regulates Notch degradation and modulates stem cell development and neurogenesis.[J]. Nat Commun, 2016, 7(1):10533.
[14] Wang L, Song G, Ming L, et al. MicroRNA-375 overexpression influences P19 cell proliferation, apoptosis and differentiation through the Notch signaling pathway[J]. Int J Mol Med, 2016, 37(1):47-55.
[15] 周娜. 脑缺血后处理对全脑缺血大鼠海马区Notch1/细胞周期通路的影响[D].唐山:华北理工大学,2016.
[16] 何砚如,陈立娟,马根山. Notch信号通路对缺血心肌的保护作用[J]. 东南大学学报:医学版,2016,35(1):139-143.
[17] 龚薇薇. INOSINE对脑缺血再灌流后脑组织Cyclin D1、CDK4表达的影响[D]. 青岛:青岛大学, 2004.
[18] 王耀琴.缺血后处理启动细胞周期促进受损心肌组织修复的研究[D].太原:山西医科大学,2013.
[19] Sriuranpong V, Borges MW, Ravi RK, et al.Notch signaling induces cell cycle arrest in small cell lung cancer cells[J]. Cancer Res, 2001, 61(7):3200-3205.
[20] 田园如画,周中和,陈会生.腺苷酸活化蛋白激酶在缺血预处理诱导的神经保护中的作用[J]. 解放军医学杂志,2015, 40(5):366-371.

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

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

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	赵利军, 李开济, 吴静, 门秀丽. 缺血后处理对大鼠肢体缺血再灌注后肾细胞凋亡的抑制作用[J]. 吉林大学学报(医学版), 2017, 43(04): 725-728.
[2]	赵利军, 李开济, 卢秋玲, 门秀丽. 缺血后处理对大鼠肢体缺血再灌注后肺损伤的保护作用及其机制[J]. 吉林大学学报(医学版), 2016, 42(02): 255-259.
[3]	齐玲, 杨阳, 刘玉翠, 朱天信, 金嵩, 臧琳, 张玉影, 吕鹏, 徐冶. 双氢青蒿素对神经母细胞瘤细胞生长的抑制作用及其机制[J]. 吉林大学学报(医学版), 2016, 42(02): 266-270.
[4]	米亚静, 刘洁, 王世伟, 王天伟, 景晓红. DNA 双加氧酶TET2在老年痴呆动物模型脑组织中的表达及其对氧化应激中神经元的保护作用[J]. 吉林大学学报(医学版), 2015, 41(04): 727-731.
[5]	刘治慧, 杨巍. 橙皮素对舌癌细胞的生长抑制作用及其机制[J]. 吉林大学学报(医学版), 2015, 41(03): 558-562.
[6]	刘颖，张萱，刘纯岩，倪文杰，毛洪涛，王珍琦. 抗精神病药奎硫平对大鼠海马神经元死亡和细胞周期的影响[J]. 吉林大学学报(医学版), 2014, 40(01): 15-18.
[7]	常明则，田晔，乔琳，狄政莉，胡彬，张茹，王虎青，吴海琴，刘勇. 葛根素预处理对大鼠局灶性脑缺血再灌注损伤的保护作用及时间窗研究[J]. 吉林大学学报(医学版), 2014, 40(01): 23-26.
[8]	刘颖，张萱，刘纯岩，倪文杰，毛洪涛，王珍琦. 抗抑郁药奥氮平对大鼠海马神经细胞凋亡和细胞周期的影响[J]. 吉林大学学报(医学版), 2013, 39(6): 1094-1097.
[9]	李悦玮\|张文琪\|孙景辉\|张科强. 尼可地尔后处理对心肌缺血-再灌注损伤大鼠心肌的保护作用[J]. J4, 2012, 38(6): 1160-1163.
[10]	王少华\|洪新雨\|薄其青\|葛鑫\|崔佳乐. 日本刺沙蚕纤溶酶对局灶性脑缺血大鼠的神经保护作用[J]. J4, 2012, 38(5): 957-960.
[11]	张萱, 刘颖, 刘纯岩, 王珍琦. 抗精神病药奎硫平的神经保护作用[J]. J4, 2011, 37(5): 779-783.
[12]	张萱, 刘纯岩, 王英男, 关松磊, 王珍琦. 抗抑郁药万拉法新对大鼠海马神经细胞凋亡和细胞周期的影响[J]. J4, 2011, 37(4): 577-581.
[13]	杨宇, 杨欣, 孙欣, 吴昊, 王全颖, 杨广笑, 吴江. 可分泌表达神经保护肽的重组慢病毒对闭合性脑损伤小鼠的神经保护作用[J]. J4, 2010, 36(4): 616-619.
[14]	吴静, 杨巍, 孙际童, 苏静波, 魏晓曦, 李一. AIRE通过调控Notch1表达影响CD4+CD25+Treg的作用及其机制[J]. J4, 2009, 35(6): 983-986.
[15]	王珍琦,张萱,李鹏武，刘扬，贾晓晶，龚守良. 抗精神病药奥氮平的神经保护作用[J]. J4, 2008, 34(4): 556-559.