[1]方立),王雪晶),荆婧),等.rhG-CSF对大鼠缺血缺氧神经元巨自噬水平的影响*[J].郑州大学学报(医学版),2013,(06):753.
 FANG Li),WANG Xuejing),JING Jing),et al.Effects of rhG-CSF on macroautophagy of ischemia hypoxia SH-SY5Y cells and rat neurons[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2013,(06):753.
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rhG-CSF对大鼠缺血缺氧神经元巨自噬水平的影响*
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2013年06期
页码:
753
栏目:
论著
出版日期:
2013-11-20

文章信息/Info

Title:
Effects of rhG-CSF on macroautophagy of ischemia hypoxia SH-SY5Y cells and rat neurons
作者:
方立1)王雪晶1)荆婧1)马耀华1)邓文静1)张雯雯1)滕军放1 2)#
1)郑州大学第一附属医院神经内科 郑州 450052;2)郑州大学第一附属医院河南省高等学校临床医学重点学科开放实验室 郑州 450052
Author(s):
FANG Li1) WANG Xuejing1) JING Jing1) MA Yaohua1) DENG Wenjing1) ZHANG Wenwen1) TENG Junfang1 2)
1)Department of Neurology, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052
关键词:
重组人粒细胞集落刺激因子 巨自噬 缺血缺氧 大鼠 SH-SY5Y
Keywords:
recombinant human granulocyte colony stimulating factor macroautophagy ischemia hypoxia rat SH-SY5Y
分类号:
R743.3
摘要:
目的:探讨重组人粒细胞集落刺激因子(rhG-CSF)对缺血缺氧神经元巨自噬水平的影响及意义。方法:采取氧糖剥夺法制作细胞缺氧模型,Western blot法检测0、100、200 μg/L rhG-CSF干预下细胞巨自噬标志微管相关蛋白轻链3膜型和胞质型比值(LC3Ⅱ/LC3Ⅰ)的变化,单丹磺酰戊二胺(MDC)法荧光观察巨自噬空泡变化,MTT法检测细胞活力。应用改良线栓法建立大鼠局灶性脑缺血模型,rhG-CSF干预前及干预7 d后进行改良的神经功能缺损评分评定,Western blot法检测干预7 d后大鼠皮层神经元LC3Ⅱ/LC3Ⅰ,免疫组化法检测LC3蛋白的表达。结果:随着rhG-CSF给药剂量的增加,细胞缺氧模型LC3Ⅱ/LC3Ⅰ和巨自噬空泡数量逐渐增加(F=288.686,154.698,P<0.001),细胞活力也相应增强(F=74.456 P<0.001)。动物实验中, 干预7 d后,rhG-CSF组大鼠缺血神经元LC3Ⅱ/LC3Ⅰ(F=573.085,P<0.001)及LC3阳性细胞数(F=88.945,P<0.001)分别较假手术组及模型组上调,大鼠的神经功能缺损评分较干预前明显降低(F组间=20.403,F时间=19.193,F交互=33.415,P<0.001)。结论:局灶性脑缺血后rhG-CSF可能通过上调巨自噬水平参与神经元的保护。
Abstract:
Aim: To explore the effect and significance of rhG-CSF on neuronal macroautophagy in hypoxia ischemia rats. Methods: Cell hypoxia model was established by taking oxygen-glucose deprivation. After dealing cells with 0,100,200 μg/L rhG-CSF, the marker of macrophage, LC3Ⅱ/LC3Ⅰ was measured by Western blot, macroautophagy vesicle change was analyzed by monodansylcadaverine(MDC), and MTT was used to investigate the cell viability. Focal cerebral ischemia rat model was established by improved suture method. The modified neurological severity score(mNSS)was adopted to evaluate neural function recovery before and after the intervention of rhG-CSF,LC3Ⅱ/LC3 Ⅰ of cortical neurons in rats was observed by Western blot, and LC3 protein expression was detected by immunohistochemistry. Results: With the increase of the dosage of rhG-CSF, the ratio of LC3Ⅱ/LC3Ⅰ and the number of macroautophagy vesicles gradually increased(F=288.686,154.698,P<0.001),and the cell viability also gradually increased(F=74.456, P<0.001). Compared with sham operation group and model group, LC3Ⅱ/LC3Ⅰ and LC3-positive cells(F=573.085 and 88.945,P<0.001)raised in rhG-CSF group. After treatment, the mNSS of rats in rhG-CSF group was lower(Fgroup=20.403,Ftime=19.193,Fmutual=33.415,P<0.001). Conclusion: rhG-CSF may participate in the neural protection after focal cerebral ischemia by raising macroautophagy activity.

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备注/Memo

备注/Memo:
*河南省基础与前沿技术研究计划项目 092102310217; 河南省医学科技攻关项目 200903037
#通讯作者, 男,1960年11月生,博士,教授,主任医师,研究方向:脑血管病的基础与临床研究,E-mail:tjf6666@yahoo.com.cn
更新日期/Last Update: 2013-11-20