大鼠外伤性癫痫模型皮层SPP1蛋白的表达
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摘要
目的检测分泌型焦磷酸蛋白1 (Secreted phosphoprotein 1,SPP1)在外伤性癫痫(post-traumaticepilepsy,PTE)大鼠皮层脑组织中的表达变化,探讨其与外伤性癫痫的关联。方法随机将18只大鼠分为正常对照组、生理盐水组和实验组。建立外伤性癫痫模型。采用免疫组化法检测GFAP蛋白,采用Western blot法检测SPP1蛋白。结果大鼠行为学平均评分为4分;EEG显示明显痫性放电;免疫组化结果显示,实验组大鼠脑组织GFAP阳性细胞与正常对照组和生理盐水组比较,显著增高(P <0.05);生理盐水组GFAP阳性细胞与正常对照组比较,差异不具有统计学意义(P> 0.05),以上结果综合表明成功建立了大鼠外伤性癫痫模型。Western blot结果显示,实验组SPP1蛋白表达量较正常对照组和生理盐水组显著增高(P <0.01);生理盐水组SPP1蛋白表达量较正常对照组不显著,差异无统计学意义(P> 0.05)。结论 SPP1在外伤性癫痫大鼠皮层脑组织中表达显著上调,外伤性癫痫的发生机制可能与SPP1的表达异常增高有关。
Objective The article is to detect the expression of Secreted phosphoprotein 1(SPP1) in cortical brain tissue of post-traumatic epilepsy(PTE) rats and to explore its relationship with PTE. Methods 18 adult rats were randomly distributed into three groups, normal control group, physiological saline group and experimental group. PTE model was established. GFAP was detected by immunohistochemistry and Western blot was used to detect SPP1 protein. Results The average behavioral score of rats was 4; EEG showed epileptiform discharge obviously.Immunohistochemistry results showed that the GFAP positive cells in the brain of the experimental group were higher than normal control and saline group(P < 0.05); Comparing the number of GFAP positive cells in saline group and that in the normal control group, the difference have no statistical significance(P > 0.05). The above results indicate that the post-traumatic epilepsy model was successfully established. Western blot results showed that protein expression of SPP1 in the experimental group was significantly higher than normal control and saline group(P < 0.01); Comparing the SPP1 protein expression level in the saline group and that in the normal control group, the difference have no statistical significance(P > 0.05). Conclusion Expression of SPP1 in cortical brain tissue of post-traumatic epilepsy rats is increased significantly, and the elevated SPP1 may be related to formation of post-traumatic epilepsy.
Objective The article is to detect the expression of Secreted phosphoprotein 1(SPP1) in cortical brain tissue of post-traumatic epilepsy(PTE) rats and to explore its relationship with PTE. Methods 18 adult rats were randomly distributed into three groups, normal control group, physiological saline group and experimental group. PTE model was established. GFAP was detected by immunohistochemistry and Western blot was used to detect SPP1 protein. Results The average behavioral score of rats was 4; EEG showed epileptiform discharge obviously.Immunohistochemistry results showed that the GFAP positive cells in the brain of the experimental group were higher than normal control and saline group(P < 0.05); Comparing the number of GFAP positive cells in saline group and that in the normal control group, the difference have no statistical significance(P > 0.05). The above results indicate that the post-traumatic epilepsy model was successfully established. Western blot results showed that protein expression of SPP1 in the experimental group was significantly higher than normal control and saline group(P < 0.01); Comparing the SPP1 protein expression level in the saline group and that in the normal control group, the difference have no statistical significance(P > 0.05). Conclusion Expression of SPP1 in cortical brain tissue of post-traumatic epilepsy rats is increased significantly, and the elevated SPP1 may be related to formation of post-traumatic epilepsy.
引文
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[2]Albertsson A M,Zhang X,Leavenworth J,et al.The effect of osteopontin and osteopontin-derived peptides on preterm brain injury[J].J Neuroinflamm,2014,11(1):197.
[3]Chan J L,Reeves T M,Phillips L L.Osteopontin expression in acute immune response mediates hippocampal synaptogenesis and adaptive outcome following cortical brain injury[J].Exp Neurol,2014,261:757-771.
[4]Ueda Y,Willmore L J,Triggs W J.Amygdalar injection of FeCl2 causes spontaneous recurrent seizures[J].Exp Neurol,1988,153(1):123-127.
[5]Glushakov A V,Glushakova O Y,DoréS,et al.Animal Models of Posttraumatic Seizures and Epilepsy[J].Methods Mol Biol,2016,1462:481-519.
[6]Short A D,Bian J,Ghosh T K,et al.Intracellular Ca2+pool content is linked to control of cell growth[J].Proc Natl Acad Sci USA,1993,90(11):4986-4990.
[7]Kumaran C,Shivakumar K.Calcium and superoxide anionmediated mitogenic action of subetance P on cardiac fibroblasts[J].Am J Physiol Heart Circ Physiol,2002,282(5):H1855-1862.
[8]Khaitin A,Rudkovskii M,Uzdensky A.Camediates axotomy-induced necrosis and apoptosis of satellite glial cells remote from the transection site in the isolated crayfish mechanoreceptor[J].Mol Cell Neurosci,2018,88:7-15.
[9]Hubbard J A,Szu J I,Yonan J M,et al.Regulation of astrocyte glutamate transporter-1(GLT1)and aquaporin-4(AQP4)expression in a model of epilepsy[J].Exp Neurol,2016,283(Pt A):85-96.
[10]Bedner P,Steinh?user C.Altered kir and gap junction channels in temporal lobe epilepsy[J].Neurochem Int,2013,63(7):682-687.
[11]Li Z,Mi X,Xiong Y,et al.Elevated Expression of the Delta-Subunit of Epithelial Sodium Channel in Temporal Lobe Epilepsy Patients and Rat Model[J].J Mol Neurosci,2015,57(4):510-518.
[12]Orr B O,Gorczyca D,Younger M A,et al.Composition and control of a Deg/ENaC channel during presynaptic homeostatic plasticity[J].Cell Rep,2017,20(8):1855-1866.