Study on the Relationship between the Expression Ratio of Induced Nitric Oxide Synthase over Glial Fibrillary Acidic Protein and the Time of Injury after Brain Concussion in Rats_West China Medical Journal

Authors：

 Dong HewenHewen , DaiHao , XiaYongjun , ZengDa , JinWei ,  LiuMin

West China School of Basic and Forensic Medicine, Sichuan University, Chengdu, Sichuan 610041, P. R. China;

Corresponding author：

LiuMin, Email: min8liu@hotmail.com

Keywords：

Induced nitric oxide synthase; Glial fibrillary acidic protein; Brain concussion; Injury time determination; Ratio of the percentage of positive cells; Rats

DOI：

10.7507/1002-0179.201600195

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Objective To study the relationship between the expression ratio of induced nitric oxide synthase (iNOS) over glial fibrillary acidic protein (GFAP) and the time of injury after brain concussion in rat, in order to acquire a new visual angle for determining injury time of cerebral concussion. Methods Eighty-five healthy Sprague-Dawley rats were divided into three groups randomly: model group (n=25), experimental group (n=55), and control group (n=5). The rats in the model group were used to confirm the attack hight to make the model of brain concussion; according to the time of execution, rats in the experimental group were then subdivided into 11 groups with 5 rats in each subgroup, and their execution time was respectively hour 0.5, 1, 3, 6, 12, 24, 48, 96, 168, 240, and 336; the rats in the control group were executed after fed for 24 hours. After the model of cerebral concussion was established through freefalling dart method, hematoxylin-eosin staining and immunohistochemistry staining of iNOS and GFAP were conducted for the brain of the rats. All related experimental results were studied by using microscope with image analytical system and homologous statistics. Results The ratio of positive expression of iNOS over that of GFAP increased gradually during hour 0.5- 3 after injury in brain (from 5.03 to 10.47). At the same time, the positive expression of iNOS increased significantly (from 14.61% to 37.45%). However, the increase of the positive expression of GFAP was not obvious. Between hour 3 and 12, the ratio began to decline to 4.98, which was still at a high level, and during the same time period, the positive expressions of iNOS and GFAP also experienced the same change pattern. Later, the ratio began to decline between hour 12 and 336 after injury (from 4.98 to 0.95). All ratios at this time were lower than those between hour 0.5 and 12. The positive expression of iNOS and GFAP both increased to a climax before declining. Conclusions The ratio of positive expression of iNOS over GFAP and the respective change pattern of iNOS and GFAP can be used as the evidence of estimating the injury time of cerebral concussion. We can use the ratio of two or more markers to provide a new visual angle for concluding the concussion injury time.

Citation： Dong HewenHewen, DaiHao, XiaYongjun, ZengDa, JinWei, LiuMin. Study on the Relationship between the Expression Ratio of Induced Nitric Oxide Synthase over Glial Fibrillary Acidic Protein and the Time of Injury after Brain Concussion in Rats. West China Medical Journal, 2016, 31(4): 718-723. doi: 10.7507/1002-0179.201600195 Copy

1.	Nylén K, Ost M, Csajbok LZ, et al. Increased serum-GFAP in patients with severe traumatic brain injury is related to outcome[J]. J Neurol Sci, 2006, 240(1/2): 85-91.
2.	Deng P, Xu HH, Zhu JZ. Preliminary study on using changes in astrocytes and neurons in the brain-stem of the rat after stab wounds for determining the time of injury[J]. Forensic Sci Int, 1999, 103(3): 181-191.
3.	王炜煜, 秦启生, 易继林. 大鼠脑挫伤后GFAP和iNOS表达与损伤时间关系的实验研究[J]. 中国法医学杂志, 2004, 19(5): 271-274.
4.	Nieto-Sampedro M, Lewis ER, Cotman CW, et al. Brain injury causes a time-dependent increase in neuronotrophic activity at the lesion site[J]. Science, 1982, 217(4562): 860-861.
5.	杨静, 王晔, 陈晓刚, 等. 大鼠脑液压冲击伤caspase-8的表达[J]. 法医学杂志, 2006, 22(1): 1-3.
6.	陈锐, 余彬华, 胡玲, 等. 大鼠脑震荡伤后脑组织HSP70、bFGF及TGF-β1表达变化[J]. 法医学杂志, 2009, 25(4): 249-253.
7.	董阳. 一重和三重脑震荡鼠BB、BW和FS行为学变化及脑组织GFAP和MBP变化研究[D]. 昆明: 昆明医科大学, 2013.
8.	Marmarou A, Foda MA, van den Brink W, et al. A new model of diffuse brain injury in rats. Part I: Pathophysiology and biomechanics[J]. J Neurosurg, 1994, 80(2): 291-300.
9.	陈锐, 严治, 胡玲, 等. 大鼠脑震荡模型建立法的改良和评价[J]. 广东医学院学报, 2010, 28(4): 362-363.
10.	Kamijo R, Harada H, Matsuyama T, et al. Requirement for transcription factor IRF-1 in NO synthase induction in macrophages[J]. Science, 1994, 263(5153): 1612-1615.
11.	Saura M, Zaragoza C, Bao G, et al. Interaction of interferon regulatory factor-1 and nuclear factor kappa B during activation of inducible nitric oxide synthase transcription[J]. J Mol Biol, 1999, 289(3): 459-471.
12.	Martin E, Nathan C, Xie Q. Role of interferon regulatory factor 1 in induction of nitric oxide synthase[J]. J Exp Med, 1994, 180(3): 977-984.
13.	Xie K, Huang S, Dong Z, et al. Destruction of bystander cells by tumor cells transfected with inducible nitric oxide (NO) synthase gene[J]. J Natl Cancer Inst, 1997, 89(6): 421-427.
14.	Robertson FM, Long BW, Tober KL, et al. Gene expression and cellular sources of inducible nitric oxide synthase during tumor promotion[J]. Carcinogenesis, 1996, 17(9): 2053-2059.
15.	Palmieri D, Capponi S, Geroldi A, et al. TNF alpha induces the expression of genes associated with endothelial dysfunction through p38MAPK-mediated down-regulation of miR-149[J]. Biochem Biophys Res Commun, 2014, 443(1): 246-251.
16.	Ono K, Suzuki H, Sawada M. Delayed neural damage is induced by iNOS-expressing microglia in a brain injury model[J]. Neurosci Lett, 2010, 473(2): 146-150.
17.	Togashi H, Sasaki M, Frohman E, et al. Neuronal (type Ⅰ) nitric oxide synthase regulates nuclear factor kappaB activity and immunologic (type Ⅱ) nitric oxide synthase expression[J]. Proc Natl Acad Sci USA, 1997, 94(6): 2676-2680.
18.	Huang D, Shenoy A, Cui J, et al. In situ detection of AP sites and DNA Strand breaks bearing 3’-phosphate termini in ischemic mouse brain[J]. FASEB J, 2000, 14(2): 407-417.
19.	Middeldorp J, Hol EM. GFAP in health and disease[J]. Prog Neurobiol, 2011, 93(3): 421-443.
20.	Hozumi I, Chiu FC, Norton WT. Biochemical and immunocytochemical changes in glial fibrillary acidic protein after stab wounds[J]. Brain Res, 1990, 524(1): 64-71.
21.	李玉红, 闫平, 孟晓萍, 等. 实验性大鼠颅脑损伤后GFAP的表达研究[J]. 伤残医学杂志, 2004, 12(2): 1-3.

1. Nylén K, Ost M, Csajbok LZ, et al. Increased serum-GFAP in patients with severe traumatic brain injury is related to outcome[J]. J Neurol Sci, 2006, 240(1/2): 85-91.
2. Deng P, Xu HH, Zhu JZ. Preliminary study on using changes in astrocytes and neurons in the brain-stem of the rat after stab wounds for determining the time of injury[J]. Forensic Sci Int, 1999, 103(3): 181-191.
3. 王炜煜, 秦启生, 易继林. 大鼠脑挫伤后GFAP和iNOS表达与损伤时间关系的实验研究[J]. 中国法医学杂志, 2004, 19(5): 271-274.
4. Nieto-Sampedro M, Lewis ER, Cotman CW, et al. Brain injury causes a time-dependent increase in neuronotrophic activity at the lesion site[J]. Science, 1982, 217(4562): 860-861.
5. 杨静, 王晔, 陈晓刚, 等. 大鼠脑液压冲击伤caspase-8的表达[J]. 法医学杂志, 2006, 22(1): 1-3.
6. 陈锐, 余彬华, 胡玲, 等. 大鼠脑震荡伤后脑组织HSP70、bFGF及TGF-β1表达变化[J]. 法医学杂志, 2009, 25(4): 249-253.
7. 董阳. 一重和三重脑震荡鼠BB、BW和FS行为学变化及脑组织GFAP和MBP变化研究[D]. 昆明: 昆明医科大学, 2013.
8. Marmarou A, Foda MA, van den Brink W, et al. A new model of diffuse brain injury in rats. Part I: Pathophysiology and biomechanics[J]. J Neurosurg, 1994, 80(2): 291-300.
9. 陈锐, 严治, 胡玲, 等. 大鼠脑震荡模型建立法的改良和评价[J]. 广东医学院学报, 2010, 28(4): 362-363.
10. Kamijo R, Harada H, Matsuyama T, et al. Requirement for transcription factor IRF-1 in NO synthase induction in macrophages[J]. Science, 1994, 263(5153): 1612-1615.
11. Saura M, Zaragoza C, Bao G, et al. Interaction of interferon regulatory factor-1 and nuclear factor kappa B during activation of inducible nitric oxide synthase transcription[J]. J Mol Biol, 1999, 289(3): 459-471.
12. Martin E, Nathan C, Xie Q. Role of interferon regulatory factor 1 in induction of nitric oxide synthase[J]. J Exp Med, 1994, 180(3): 977-984.
13. Xie K, Huang S, Dong Z, et al. Destruction of bystander cells by tumor cells transfected with inducible nitric oxide (NO) synthase gene[J]. J Natl Cancer Inst, 1997, 89(6): 421-427.
14. Robertson FM, Long BW, Tober KL, et al. Gene expression and cellular sources of inducible nitric oxide synthase during tumor promotion[J]. Carcinogenesis, 1996, 17(9): 2053-2059.
15. Palmieri D, Capponi S, Geroldi A, et al. TNF alpha induces the expression of genes associated with endothelial dysfunction through p38MAPK-mediated down-regulation of miR-149[J]. Biochem Biophys Res Commun, 2014, 443(1): 246-251.
16. Ono K, Suzuki H, Sawada M. Delayed neural damage is induced by iNOS-expressing microglia in a brain injury model[J]. Neurosci Lett, 2010, 473(2): 146-150.
17. Togashi H, Sasaki M, Frohman E, et al. Neuronal (type Ⅰ) nitric oxide synthase regulates nuclear factor kappaB activity and immunologic (type Ⅱ) nitric oxide synthase expression[J]. Proc Natl Acad Sci USA, 1997, 94(6): 2676-2680.
18. Huang D, Shenoy A, Cui J, et al. In situ detection of AP sites and DNA Strand breaks bearing 3’-phosphate termini in ischemic mouse brain[J]. FASEB J, 2000, 14(2): 407-417.
19. Middeldorp J, Hol EM. GFAP in health and disease[J]. Prog Neurobiol, 2011, 93(3): 421-443.
20. Hozumi I, Chiu FC, Norton WT. Biochemical and immunocytochemical changes in glial fibrillary acidic protein after stab wounds[J]. Brain Res, 1990, 524(1): 64-71.
21. 李玉红, 闫平, 孟晓萍, 等. 实验性大鼠颅脑损伤后GFAP的表达研究[J]. 伤残医学杂志, 2004, 12(2): 1-3.

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Study on the Relationship between the Expression Ratio of Induced Nitric Oxide Synthase over Glial Fibrillary Acidic Protein and the Time of Injury after Brain Concussion in Rats

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