单纯性脑震荡对大鼠海马肿瘤坏死因子表达的影响
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摘要
目的观察单纯性脑震荡(PCC)大鼠海马肿瘤坏死因子(TNF-α)表达的变化。方法应用金属单摆闭合脑损伤打击装置制作清醒状态下PCC大鼠模型,将大鼠随机分为PCC 3 h、12 h、1 d、2 d、3 d、7 d亚组,每个亚组5只;另设正常对照组(n=5)。采用TNF-α多克隆抗体进行免疫组织化学染色和Western blotting实验,观察PCC组和正常对照组大鼠海马脑区的TNF-α免疫阳性表达以及TNF-α蛋白表达。结果正常对照组大鼠海马CA1、CA2、CA3、CA4、DG1和DG2中,TNF-α阳性产物的表达均较弱,阳性细胞轮廓不清且数量少;PCC组TNF-α阳性产物表达和阳性细胞数量随时间推移逐渐增加,3 d达高峰,7 d下降。PCC不同时间点亚组各脑区TNF-α的表达均显著高于正常对照组(均P<0.05)。PCC组各亚组TNF-α阳性细胞计数及蛋白水平均显著高于正常对照组(均P<0.05)。结论 TNF-α在PCC大鼠海马致伤早期呈现表达增高趋势,在脑损伤继发性病理变化中起重要作用。
Objective To observe the expression of tumor necrosis factor alpha( TNF-α) in the hippocampus of rats with pure cerebral concussion( PCC). Methods The rat PCC experimental model was made by metallic pendulum-striker closed-head concussive equipment during the wake state. PCC rats were randomly divided into 3 h,12 h,1 d,2 d,3 d,7 d subgroups,5 rats for each subgroup; in addition,normal control group( n = 5) was set.The immuno-expression and protein expression of TNF-α in hippocampus of rats were evaluated by immunohistochemistry and Western blotting. Results The positive expression of TNF-α was weak in CA1,CA2,CA3,CA4,DG1 and DG2 areas of hippocampus in normal control group; the positive cell outline was not clear,and positive cell was less. With the time coursing,the expression of TNF-α and positive cell number in PCC group were progressively increased peaking at 3 d and decreasing at 7 d. The positive expression of TNF-α of brain areas in PCC subgroups at different time points were significantly higher than that in normal control group( all P < 0. 05). The number of TNF-α positive cell and TNF-α protein level in each subgroup of PCC group were significantly higher than those in control group( all P < 0. 05). Conclusion TNF-α shows an increasing expression in the hippocampus during the early stage of PCC,and plays an important role in secondary pathological changes of the brain injury.
Objective To observe the expression of tumor necrosis factor alpha( TNF-α) in the hippocampus of rats with pure cerebral concussion( PCC). Methods The rat PCC experimental model was made by metallic pendulum-striker closed-head concussive equipment during the wake state. PCC rats were randomly divided into 3 h,12 h,1 d,2 d,3 d,7 d subgroups,5 rats for each subgroup; in addition,normal control group( n = 5) was set.The immuno-expression and protein expression of TNF-α in hippocampus of rats were evaluated by immunohistochemistry and Western blotting. Results The positive expression of TNF-α was weak in CA1,CA2,CA3,CA4,DG1 and DG2 areas of hippocampus in normal control group; the positive cell outline was not clear,and positive cell was less. With the time coursing,the expression of TNF-α and positive cell number in PCC group were progressively increased peaking at 3 d and decreasing at 7 d. The positive expression of TNF-α of brain areas in PCC subgroups at different time points were significantly higher than that in normal control group( all P < 0. 05). The number of TNF-α positive cell and TNF-α protein level in each subgroup of PCC group were significantly higher than those in control group( all P < 0. 05). Conclusion TNF-α shows an increasing expression in the hippocampus during the early stage of PCC,and plays an important role in secondary pathological changes of the brain injury.
引文
[1]Ferguson RJ,Mittenberg W,Barone DF,et al.Postconcussion syndrome following sports-related head injury:expectation as etiology[J].Neuropsychology,1999,13:582.
[2]Chiang CK,Huang JW,Hsu SP,et al.Plasma interleukin-18 levels in hemodialysis patients:increased by dialysis process and association with interleukin-6 and tumor necrotic factor-alpha[J].Blood Purif,2006,24:174.
[3]于建云,李树华,韩晓华,等.脑震荡鼠细胞组织病理学研究[J].昆明医学院学报,2003,24:1.
[4]于建云,林海英,吴春云,等.大鼠脑震荡后8天不同损伤程度空间认知行为的变化[J].昆明医学院学报,2005,26:1.
[5]于建云,李俊祥,郭泽云,等.实验大鼠轻中型闭合性脑损伤昏迷指标与分级标准[J].法医学杂志,2008,1:8.
[6]Villoslada P,Genain CP.Role of nerve growth factor and other trophic factors in brain inflammation[J].Prog Brain Res,2004,146:403.
[7]江基尧.现代颅脑损伤学[M].第3版.上海:第二军医大学出版社,2010:625-627.
[8]Suzuki S,Tanaka K,Suzuki N.Ambivalent aspects of interleukin-6in cerebral ischemia:inflammatory versus neurotrophic aspects[J].J Cereb Blood Flow Metab,2009,29:464.
[9]Ashdown H,Joita S,Luheshi GN,et al.Acute brain cytokine responses after global birth hypoxia in the rat[J].J Neurosci Res,2008,86:3401.
[10]李玉环,吴德野,董世峰,等.TNF-α在单纯性脑震荡大鼠皮质中的表达变化[J].昆明医科大学学报,2015,36:4.
[11]严治,孙小丽,胡玉莲,等.大鼠脑挫伤后脑与其他器官组织中TNF-α表达的比较[J].法医学杂志,2012,28:261.
[12]刘盛明,游潮,候敏.创伤性脑损伤脑组织NF-κB、TNF-α和血清IL-6表达的实验研究[J].四川医学,2007,28:1254.
[13]张援平,张锦泉,王安平,等.脑震荡早期血清TNF-α和IL-1β的变化[J].沈阳部队医药,2007,20:27.
[14]Lynch MA.Long-term potentiation and memory[J].Physiol Rev,2004,84:87.
[15]Li P,Jiang H,Wu H,et al.AH6809 decreases production of inflammatory mediators by PGE2-EP2-c AMP signaling pathway in an experimentally induced pure cerebral concussion in rats[J].Brain Res,2018,1698:11.
[16]林海英,于建云,赵小林,等.大鼠单纯性脑震荡损伤后不同时期空间学习记忆功能的研究[J].中国行为医学科学,2006,15:769.
[17]Barth AM,Mody I.Changes in hippocampal neuronal activity during and after unilateral selective hippocampal ischemia in vivo[J].J Neurosci,2011,31:851.
[18]Cumiskey D,Butler MP,Moynagh PN,et al.Evidence for a role for the group I metabotropic glutamate receptor in the inhibitory effect of tumor necrosis factor-alpha on long-term potentiation[J].Brain Res,2007,1136:13.
[19]Murray CA,Lynch MA.Evidence that increased hippocampal expression of the cytokine interleukin-1 beta is a common trigger for age-and stress-induced impairments in long-term potentiation[J].J Neurosci,1998,18:2974.
[2]Chiang CK,Huang JW,Hsu SP,et al.Plasma interleukin-18 levels in hemodialysis patients:increased by dialysis process and association with interleukin-6 and tumor necrotic factor-alpha[J].Blood Purif,2006,24:174.
[3]于建云,李树华,韩晓华,等.脑震荡鼠细胞组织病理学研究[J].昆明医学院学报,2003,24:1.
[4]于建云,林海英,吴春云,等.大鼠脑震荡后8天不同损伤程度空间认知行为的变化[J].昆明医学院学报,2005,26:1.
[5]于建云,李俊祥,郭泽云,等.实验大鼠轻中型闭合性脑损伤昏迷指标与分级标准[J].法医学杂志,2008,1:8.
[6]Villoslada P,Genain CP.Role of nerve growth factor and other trophic factors in brain inflammation[J].Prog Brain Res,2004,146:403.
[7]江基尧.现代颅脑损伤学[M].第3版.上海:第二军医大学出版社,2010:625-627.
[8]Suzuki S,Tanaka K,Suzuki N.Ambivalent aspects of interleukin-6in cerebral ischemia:inflammatory versus neurotrophic aspects[J].J Cereb Blood Flow Metab,2009,29:464.
[9]Ashdown H,Joita S,Luheshi GN,et al.Acute brain cytokine responses after global birth hypoxia in the rat[J].J Neurosci Res,2008,86:3401.
[10]李玉环,吴德野,董世峰,等.TNF-α在单纯性脑震荡大鼠皮质中的表达变化[J].昆明医科大学学报,2015,36:4.
[11]严治,孙小丽,胡玉莲,等.大鼠脑挫伤后脑与其他器官组织中TNF-α表达的比较[J].法医学杂志,2012,28:261.
[12]刘盛明,游潮,候敏.创伤性脑损伤脑组织NF-κB、TNF-α和血清IL-6表达的实验研究[J].四川医学,2007,28:1254.
[13]张援平,张锦泉,王安平,等.脑震荡早期血清TNF-α和IL-1β的变化[J].沈阳部队医药,2007,20:27.
[14]Lynch MA.Long-term potentiation and memory[J].Physiol Rev,2004,84:87.
[15]Li P,Jiang H,Wu H,et al.AH6809 decreases production of inflammatory mediators by PGE2-EP2-c AMP signaling pathway in an experimentally induced pure cerebral concussion in rats[J].Brain Res,2018,1698:11.
[16]林海英,于建云,赵小林,等.大鼠单纯性脑震荡损伤后不同时期空间学习记忆功能的研究[J].中国行为医学科学,2006,15:769.
[17]Barth AM,Mody I.Changes in hippocampal neuronal activity during and after unilateral selective hippocampal ischemia in vivo[J].J Neurosci,2011,31:851.
[18]Cumiskey D,Butler MP,Moynagh PN,et al.Evidence for a role for the group I metabotropic glutamate receptor in the inhibitory effect of tumor necrosis factor-alpha on long-term potentiation[J].Brain Res,2007,1136:13.
[19]Murray CA,Lynch MA.Evidence that increased hippocampal expression of the cytokine interleukin-1 beta is a common trigger for age-and stress-induced impairments in long-term potentiation[J].J Neurosci,1998,18:2974.