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中华神经创伤外科电子杂志 ›› 2018, Vol. 04 ›› Issue (03) : 161 -166. doi: 10.3877/cma.j.issn.2095-9141.2018.03.008

所属专题: 文献

基础研究

NKCC1、KCC2及mTOR相关蛋白4E-BP1在外伤性癫痫中的表达及意义
魏书山1, 王丰1, 林元相1,(), 康德智1, 黄小芬1, 余良宏1, 林章雅1, 张聪1   
  1. 1. 350005 福州,福建医科大学附属第一医院神经外科
  • 收稿日期:2017-12-01 出版日期:2018-06-15
  • 通信作者: 林元相
  • 基金资助:
    福建省自然科学基金项目(2015J0105); 福建省医学创新基金(2014-CXB-14)

Expression and significance of NKCC1, KCC2 and mTOR pathway associated protein 4E-BP1 in post-traumatic epilepsy

Shushan Wei1, Feng Wang1, Yuanxiang Lin1,(), Dezhi Kang1, Xiaofen Huang1, Lianghong Yu1, Zhangya Lin1, Cong Zhang1   

  1. 1. Deparment of Neurosergery, The First Affiliated Hospital of Fujian Medical University, Fujian 350005, China
  • Received:2017-12-01 Published:2018-06-15
  • Corresponding author: Yuanxiang Lin
  • About author:
    Corresponding author: Lin Yuanxiang, Email:
引用本文:

魏书山, 王丰, 林元相, 康德智, 黄小芬, 余良宏, 林章雅, 张聪. NKCC1、KCC2及mTOR相关蛋白4E-BP1在外伤性癫痫中的表达及意义[J]. 中华神经创伤外科电子杂志, 2018, 04(03): 161-166.

Shushan Wei, Feng Wang, Yuanxiang Lin, Dezhi Kang, Xiaofen Huang, Lianghong Yu, Zhangya Lin, Cong Zhang. Expression and significance of NKCC1, KCC2 and mTOR pathway associated protein 4E-BP1 in post-traumatic epilepsy[J]. Chinese Journal of Neurotraumatic Surgery(Electronic Edition), 2018, 04(03): 161-166.

目的

探讨主要表达于神经元细胞膜上的NA+-K+-2CL-转运体(NKCC1)、K+-CL-转运体(KCC2)及表达于神经元细胞质中mTOR通路信号蛋白4E结合蛋白1(4E-BP1)在外伤性癫痫(PTE)癫痫灶中的表达及临床意义。

方法

收集2010年1月至2015年12月福建医科大学附属第一医院神经外科外伤性癫痫患者术后脑组织标本14例作为实验组;选取脑外伤患者行减压或清创手术获取的和各种病变患者手术入路不可避免要切除的正常脑组织8例作为对照组。用Western blot和实时定量荧光PCR(RT-PCR)检测14例PTE癫痫脑组织、8例对照组"正常脑组织"NKCC1、KCC2、4E-BP1的表达。

结果

Western blot显示PTE病灶脑组织中4E-BP1、NKCC1相对灰度值(0.61±0.12、0.92±0.19)高于正常脑组织(0.27±0.05、0.67±0.66),差异有统计学意义(P<0.05)。PTE病灶脑组织中KCC2相对灰度值(0.58±0.99)低于正常脑组织(0.72±0.06),差异有统计学意义(P<0.05)。RT-PCR结果显示,PTE病灶脑组织中4E-BP1、NKCC1相对灰度值(30.84±1.32、27.81±1.92)高于正常脑组织(26.94±1.24、23.52±0.74),差异有统计学意义(P<0.05)。PTE病灶脑组织中KCC2相对灰度值(21.55±1.01)低于正常脑组织(24.59±1.02),差异有统计学意义(P<0.05)。PTE组中NKCC1/KCC2比值(1.29±0.11)高于对照组(0.96±0.26),差异有统计学意义(P<0.05)。

结论

NKCC1、KCC2和mTOR通路信号蛋白4E-BP1的异常改变,可能是外伤后脑组织组织学改变及反复癫痫发作的重要分子机制。

Objective

To investigagte the expression and significance of NA+-K+-2CL- co-transporter (NKCC1), K+-CL- co-transporter (KCC2), which are located on the neuronal membrane, and mammalian target of rapamycin (mTOR) associated protein 4E blinding protein 1 (4E-BP1), which is located in cytoplasm of neuron, in post-traumatic epilepsy (PTE).

Methods

To collect 14 postoperative brain tissue specimens of post-traumatic epilepsy patients admitted to our hospital from 2010 to 2015 as the experiment group; 8 cases of normal brain tissue as the control group. And detect the expression of NKCC1, KCC2 and 4E-BP1 both in 14 cases of PTE tissue and 8 cases of normal brain tissue by Western blot and RT-PCR.

Results

The content of NKCC1 and 4E-BP1 in PTE was higher than that in normal brain tissue (P<0.05) by Western blot and RT-PCR; while the content of KCC2 was opposite (P<0.05). Besides, in PTE the ratio of NKCC1 to KCC2 was higher (1.29±0.11), compared to normal brain tissue (0.96±0.26) (P<0.05).

Conclusion

Abnormal expression of NKCC1, KCC2 and mTOR associated protein 4E-BP1 may be an important molecular mechanism of the histological changes of brain, after traumatic brain injured, and repeated seizures.

表1 RT-PCR检测所用的引物及内参照β-actin引物序列
图1 基因溶解曲线图
图2 基因扩增曲线图
图3 Wertern blot检测β-actin、4E-BP1、NKCC1、KCC2的表达
图4 PTE组和对照组NKCC1/KCC2比值图
表2 内参蛋白β-actin、4E-BP1、NKCC1和KCC2相对灰度值对比
图5 RT-PCR检测目的基因β-actin4E-BP1NKCC1KCC2表达灰度值
图6 PTE组和对照组NKCC1/KCC2比值
表3 PTE组和对照组4E-BP1相对灰度值
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