Study on the expression of NGB in hippocampus after status epliepticus in rats_Journal of Epilepsy

Authors：

ZHANG Feifei , CHENG Yanwei , Yu Minmin ,  SHI Xiangqun

Department of Neurology, General Hospital of Lanzhou Military Command, Lanzhou 730050, China;

Corresponding author：

SHI Xiangqun, Email: shixq_2003@163.com

Keywords：

Status epilepticus; Neuroglobin; Hippocampus; Rats; Experimental research

DOI：

10.7507/2096-0247.20170033

Video：

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ObjectiveTo observe the dynamic changes of neuroglobin (NGB) expression in hippocampus after status epilepticus(SE) in rats, and to explore the role of NGB in epileptic seizures.Methods40 healthy male Sprague Dawley rats were randomly divided into two group according to random number table method:control group (n=5) and epilepsy model group(n=35).Epilepsy model group according to observation time was divided into:0h, 1h, 3h, 12h, 24h, 10d and 30d.Intraperitoneal injection Lithium-pilocarpine (20 mg/kg~127 mg/kg, Li-PC) to establish the rat model of SE.Observe the behavioral changes in rats with epilepsy.Nissl staining was used to detect the neuronal damage in hippocampus. Streptavidin-biotin-peroxidase complex immunohistochemical method was used to detect the expression level of NGB in hippocampus;ResultsAfter SE, the neurons in hippocampus were severely damaged with the progress of epileptic seizures, the number of surviving neurons in CA1, CA3 regions showed a near linear decline.Among them, the number of surviving neurons in (12h, 24h, 10d, 30d)CA1, (0h, 12h, 24h, 10d, 30d)CA3 and(12h, 24h, 10d, 30d) DG area were significantly lower than that of the control group (P < 0.05).The expression level of NGB in CA1, CA3 and DG region of hippocampus were increased after SE, and both of CA1 and DG were reached peak in 24h after SE, but was still higher than the control group.And the CA3 area showed a continue rising trend.Among them, CA1(24h, 10d, 30d), CA3(24h, 10d, 30d) and DG(12h, 24h, 10d, 30d) were higher than that of control group significantly (P < 0.05).In addition, it was found that there was a positive correlation between the number of surviving neurons in CA3 area and the expression level of NGB (R=0.306, P=0.011).ConclusionUp-regulation of NGB expression in hippocampus after status epilepticus, and was positively correlated with the number of neurons in the CA3 area, suggesting that up regulation of NGB expression may be a compensatory protective mechanism of ischemic injury induced by seizures, and participate in the protection of epilepsy related neuronal damage.

Citation： ZHANG Feifei, CHENG Yanwei, Yu Minmin, SHI Xiangqun. Study on the expression of NGB in hippocampus after status epliepticus in rats. Journal of Epilepsy, 2017, 3(3): 217-222. doi: 10.7507/2096-0247.20170033 Copy

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8.	Li RC, Lee SK, Pouranfar F, et al. Hypoxia diferentially regulates the expression of neuroglobin and cytoglobin in rat brain. Brain Research, 2006, 1096(1): 173-179.
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11.	Shang A, Zhou D, Wang L, et al. Increased neuroglobin levels in the cerebral cortex and serum after ischemia-reperfusion insults. Brain Res, 2006, 1078(10): 219-226.
12.	Jixue Yang, Fang He, Qiang Meng, et al. Inhibiting HIF-1α decreases expression of TNF-α and Caspase-3 in specific brain regions exposed kainic acid-induced statusepilepticus. Cell Physiol Biochem, 2016, 38(2): 75-82.
13.	Ye SQ, Zhou XY, Lai XJ, et al. Silencing neuroglobin enhances neuronal vulnerability toxidative injury by down-regulating 14-3-3 gamma. Acta Pharmacol Sin, 2009, 30(7): 913-918.
14.	刘新建, 刘丽娜, 温慧敏, 等.缺氧缺血性脑损伤新生大鼠海马区脑红蛋白、凋亡相关蛋白Bcl-2、Bax的表达及其相关性.细胞与分子免疫学杂志, 2010, 26(10): 1040-1042.

1. Zhanyang Yu, Ning Liu, Jianxiang Liu, et al. Neuroglobin, a novel target for endogenous neuroprotection against stroke and neurodegenerative disorders. Int J Mol SCI, 2012, 13(7): 6995-7014.
2. Sun Y J, Jin K L, Peel A, et al. Neuroglobin protects the brain from experimental stroke in vivo. Proc Natl Acad Sci, 2003, 100(11): 3497-3500.
3. Racine RJ, Gartner JG, Burnham WM. Epileptiform activity and neural plasticity in limbic structures. Brain Res, 1972, 47(4): 262-268.
4. Paxinos G, Watson C. The rat brain in Stereotaxic coordinates. Academic press, 1986, 3(2): 6.
5. 何华.如何正确的对免疫组化结果进行半定量评分.中外健康文摘, 2008, 5(7):45.
6. Burmester T, Weich B, Reinhardt S, et al. A vertebrate globin expressed in the brain. Nature, 2000, 407(10):520-523.
7. Xiameng Chen, Yuan Liu, Lin Zhang, et al. Long-term neuroglobin expression of human astrocytes following brain trauma. Neuroscience Letters, 2015, 606(4): 194-199.
8. Li RC, Lee SK, Pouranfar F, et al. Hypoxia diferentially regulates the expression of neuroglobin and cytoglobin in rat brain. Brain Research, 2006, 1096(1): 173-179.
9. 谢金祥.高压氧综合治疗癫痢的经验.中华理疗杂志, 1999, 22(5): 338-340.
10. Li SQ, Li WB, Zhang M, et al. The role of neuroglobin in the neuroprotection of limb ischemic preconditioning in rats. Mol Neurobiol, 2013, 47(2): 197-208.
11. Shang A, Zhou D, Wang L, et al. Increased neuroglobin levels in the cerebral cortex and serum after ischemia-reperfusion insults. Brain Res, 2006, 1078(10): 219-226.
12. Jixue Yang, Fang He, Qiang Meng, et al. Inhibiting HIF-1α decreases expression of TNF-α and Caspase-3 in specific brain regions exposed kainic acid-induced statusepilepticus. Cell Physiol Biochem, 2016, 38(2): 75-82.
13. Ye SQ, Zhou XY, Lai XJ, et al. Silencing neuroglobin enhances neuronal vulnerability toxidative injury by down-regulating 14-3-3 gamma. Acta Pharmacol Sin, 2009, 30(7): 913-918.
14. 刘新建, 刘丽娜, 温慧敏, 等.缺氧缺血性脑损伤新生大鼠海马区脑红蛋白、凋亡相关蛋白Bcl-2、Bax的表达及其相关性.细胞与分子免疫学杂志, 2010, 26(10): 1040-1042.

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Study on the expression of NGB in hippocampus after status epliepticus in rats

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