颅脑创伤后期大鼠代谢型谷氨酸受体5表达变化及牛磺酸治疗作用
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摘要
目的探讨颅脑创伤后期(第7天)大鼠脑组织代谢型谷氨酸受体5(m Glu R5)表达变化,以及牛磺酸治疗作用。方法液压脑损伤打击仪制备液压打击颅脑创伤大鼠模型,采用随机数字表法将30只无特定病原体级Sprague-Dawley大鼠随机分为对照组、颅脑创伤组和牛磺酸治疗组(各10只),干湿重法检测大鼠脑组织含水量,实时荧光定量聚合酶链反应和Western blotting法检测水通道蛋白4(AQP4)和m Glu R5 m RNA和蛋白表达变化。结果与对照组相比,颅脑创伤组大鼠脑组织含水量(t=4.893,P=0.002)、AQP4 m RNA(t=6.523,P=0.000)和蛋白(t=4.366,P=0.008)表达水平升高,m Glu R5m RNA(t=5.776,P=0.001)和蛋白(t=3.945,P=0.014)表达水平降低;经牛磺酸治疗后,大鼠脑组织含水量(t=2.151,P=0.140)、AQP4 m RNA(t=1.144,P=0.432)和蛋白(t=0.367,P=0.804)降至正常水平,m Glu R5 m RNA(t=1.824,P=0.216)和蛋白(t=1.185,P=0.414)升至正常水平。相关分析显示,脑组织含水量与m Glu R5 m RNA(r=-0.617,P=0.014)和蛋白(r=-0.665,P=0.007)呈负相关,与AQP4蛋白呈正相关(r=0.658,P=0.008)。结论牛磺酸可以升高颅脑创伤后期(第7天)大鼠脑组织m Glu R5表达水平,降低脑水肿和脑组织含水量,具有抑制性神经递质作用。
Objective To investigate the change of metabotropic glutamate receptor 5(m Glu R5)expression level in rats with late-stage(the 7th day) traumatic brain injury(TBI) and the role of taurine.Methods The left cerebral TBI rat models were made by using lateral fluid percussion method. A total of30 specific pathogen free(SPF) male Sprague-Dawley(SD) rats were randomly divided into 3 groups: shamoperation group(control group), TBI model group(TBI group) and taurine treatment group(taurine group).Wet and dry weight method was used to measure the brain water content. Real- time fluorescentquantitative polymerase chain reaction(PCR) and Western blotting were used to detect the change of m RNAand protein expression of aquaporin 4(AQP4) and m Glu R5 in each group. Results Compared withcontrol group, the brain water content(t = 4.893, P = 0.002), AQP4 m RNA(t = 6.523, P = 0.000) and protein(t = 4.366, P = 0.008) expression were upregulated, while m Glu R5 m RNA(t = 5.776, P = 0.001) and protein(t = 3.945, P = 0.014) expression were downregulated in TBI group. After taurine treatment, the brain watercontent(t = 2.151, P = 0.140), AQP4 m RNA(t = 1.144,P = 0.432) and protein(t = 0.367, P = 0.804)decreased to normal, while m Glu R5 m RNA(t = 1.824, P = 0.216) and protein(t = 1.185, P = 0.414) increasedto normal. Correlation analysis showed brain water content was negatively correlated with m Glu R5 m RNA(r =- 0.617, P = 0.014) and m Glu R5 protein(r =- 0.665, P = 0.007), while it was positively correlated withAQP4 protein(r = 0.658, P = 0.008). Conclusions Taurine can significantly increase the m Glu R5 expression level of brain issue in the late-stage(the 7th day) of TBI and decline brain edema and brainwater content. It may be a potential protective agent as an inhibitory neurotransmitter.
Objective To investigate the change of metabotropic glutamate receptor 5(m Glu R5)expression level in rats with late-stage(the 7th day) traumatic brain injury(TBI) and the role of taurine.Methods The left cerebral TBI rat models were made by using lateral fluid percussion method. A total of30 specific pathogen free(SPF) male Sprague-Dawley(SD) rats were randomly divided into 3 groups: shamoperation group(control group), TBI model group(TBI group) and taurine treatment group(taurine group).Wet and dry weight method was used to measure the brain water content. Real- time fluorescentquantitative polymerase chain reaction(PCR) and Western blotting were used to detect the change of m RNAand protein expression of aquaporin 4(AQP4) and m Glu R5 in each group. Results Compared withcontrol group, the brain water content(t = 4.893, P = 0.002), AQP4 m RNA(t = 6.523, P = 0.000) and protein(t = 4.366, P = 0.008) expression were upregulated, while m Glu R5 m RNA(t = 5.776, P = 0.001) and protein(t = 3.945, P = 0.014) expression were downregulated in TBI group. After taurine treatment, the brain watercontent(t = 2.151, P = 0.140), AQP4 m RNA(t = 1.144,P = 0.432) and protein(t = 0.367, P = 0.804)decreased to normal, while m Glu R5 m RNA(t = 1.824, P = 0.216) and protein(t = 1.185, P = 0.414) increasedto normal. Correlation analysis showed brain water content was negatively correlated with m Glu R5 m RNA(r =- 0.617, P = 0.014) and m Glu R5 protein(r =- 0.665, P = 0.007), while it was positively correlated withAQP4 protein(r = 0.658, P = 0.008). Conclusions Taurine can significantly increase the m Glu R5 expression level of brain issue in the late-stage(the 7th day) of TBI and decline brain edema and brainwater content. It may be a potential protective agent as an inhibitory neurotransmitter.
引文
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[12]Dixon CE,Lyeth BG,Povlishock JT,Findling RL,Hamm RJ,Marmarou A,Young HE,Hayes RL.A fluid percussion model of experimental brain injury in the rat.J Neurosurg,1987,67:110-119.
[13]Wang JW,Wang HD,Zhong WZ,Li N,Cong ZX.Expression and cell distribution of metabotropic glutamate receptor 5 in the rat cortex following traumatic brain injury.Brain Res,2012,1464:73-81.
[14]Li AL.A study of cell membrane damage after severe traumatic brain injury and the effect of hypothermia on it.Tianjin:Tianjin Medical University,2006.[李爱林.重型脑创伤后细胞膜损伤及亚低温对其影响的研究.天津:天津医科大学,2006.]
[15]Fei Z,Zhang X,He YD,Li ZG,Liang JW,Yin C.Change of metabotropic glutamate receptor subtype 5 mRNA levels in diffuse brain injury with secondary insults and effect of antagonist MCPG.Zhonghua Chuang Shang Za Zhi,2001,17:199-201.[费舟,章翔,何远东,李志刚,梁景文,尹畅.二次脑损伤后鼠脑皮层代谢性谷氨酸受体5mRNA表达改变及α-甲基-4-羧基苯丙氨酸的治疗作用.中华创伤杂志,2001,17:199-201.]
[16]Wang JW,Wang HD,Cong ZX,Zhang XS,Zhou XM,Zhang DD.Activation of metabotropic glutamate receptor 5 reduces the secondary brain injury after traumatic brain injury in rats.Biochem Biophys Res Commun,2013,430:1016-1021.
[17]Leon R,Wu H,Jin Y,Wei J,Buddhala C,Prentice H,Wu JY.Protective function of taurine in glutamate-induced apoptosis in cultured neurons.J Neurosci Res,2009,87:1185-1194.
[18]Wang JW,Wang HD.The research progress of metabotropic glutamate receptor 5 in neurogliocyte.Zhonghua Shen Jing Wai Ke Ji Bing Yan Jiu Za Zhi,2013,12:470-471.[王嘉炜,王汉东.代谢性谷氨酸受体5在神经胶质细胞中的研究进展.中华神经外科疾病研究杂志,2013,12:470-471.]
[2]Liu GQ,Meng Y,Yuan YD,Cao Y,Jing M,Feng JY.Experimental study on the effect of moschus combined with borneol on cerebral edema and expression of AQP-4 after traumatic brain injury in rats.Wu Jing Hou Qin Xue Yuan Xue Bao(Yi Xue Ban),2012,21:775-777.[刘刚强,孟阳,苑亚东,曹勇,靖明,冯金勇.麝香配伍冰片对颅脑损伤大鼠脑组织含水量及水通道蛋白-4表达的影响.武警后勤学院学报(医学版),2012,21:775-777.]
[3]Haldipur P,Dupuis N,Degos V,Moniaux N,Chhor V,Rasika S,Schwendimann L,le Charpentier T,Rougier E,Amouyal P,Amouyal G,Dournaud P,Brechot C,El Ghouzzi V,Faivre J,Fleiss B,Mani S,Gressens P.HIP/PAP prevents excitotoxic neuronal death and promotes plasticity.Ann Clin Transl Neurol,2014,1:739-754.
[4]Jansson LC,Akerman KE.The role of glutamate and its receptors in the proliferation,migration,differentiation and survival of neural progenitor cells.J Neural Transm(Vienna),2014,121:819-836.
[5]Kumar A,Dhull DK,Mishra PS.Therapeutic potential of mGluR5 targeting in Alzheimer's disease.Front Neurosci,2015,9:215.
[6]Jansson LC,Louhivuori L,Wigren HK,Nordstrom T,Louhivuori V,Castren ML,Akerman KE.Effect of glutamate receptor antagonists on migrating neural progenitor cells.Eur J Neurosci,2013,37:1369-1382.
[7]Guimaraes IM,Carvalho TG,Ferguson SS,Pereira GS,Ribeiro FM.The metabotropic glutamate receptor 5 role on motor behavior involves specific neural substrates.Mol Brain,2015,8:24.
[8]Vella J,Zammit C,Di Giovanni G,Muscat R,Valentino M.The central role of aquaporins in the pathophysiology of ischemic stroke.Front Cell Neurosci,2015,9:108.
[9]Gebara E,Udry F,Sultan S,Toni N.Taurine increases hippocampal neurogenesis in aging mice.Stem Cell Res,2015,14:369-379.
[10]Zhang XB,Huang HL,Chang XL,Cai Y,Yao X.The role of taurine in treating encephaledema of rats with acute severe traumatic brain injury.Zhonghua Chuang Shang Za Zhi,2010,26:941-943.[张学斌,黄慧玲,常小丽,蔡英,姚鑫.牛磺酸对急性重型颅脑创伤大鼠脑水肿的作用.中华创伤杂志,2010,26:941-943.]
[11]Cai Y,Huang HL,Fan WJ,Wu QL,Li XQ,Su YH,Wen XC.Protection role of taurine transporter in rats brain edema followed severe traumatic head injury.Tianjin Yi Yao,2015,43:607-611.[蔡英,黄慧玲,范维佳,武俏丽,李晓茜,苏彦华,温晓昶.重型颅脑创伤大鼠后期脑水肿研究及牛磺酸转运体的作用研究.天津医药,2015,43:607-611.]
[12]Dixon CE,Lyeth BG,Povlishock JT,Findling RL,Hamm RJ,Marmarou A,Young HE,Hayes RL.A fluid percussion model of experimental brain injury in the rat.J Neurosurg,1987,67:110-119.
[13]Wang JW,Wang HD,Zhong WZ,Li N,Cong ZX.Expression and cell distribution of metabotropic glutamate receptor 5 in the rat cortex following traumatic brain injury.Brain Res,2012,1464:73-81.
[14]Li AL.A study of cell membrane damage after severe traumatic brain injury and the effect of hypothermia on it.Tianjin:Tianjin Medical University,2006.[李爱林.重型脑创伤后细胞膜损伤及亚低温对其影响的研究.天津:天津医科大学,2006.]
[15]Fei Z,Zhang X,He YD,Li ZG,Liang JW,Yin C.Change of metabotropic glutamate receptor subtype 5 mRNA levels in diffuse brain injury with secondary insults and effect of antagonist MCPG.Zhonghua Chuang Shang Za Zhi,2001,17:199-201.[费舟,章翔,何远东,李志刚,梁景文,尹畅.二次脑损伤后鼠脑皮层代谢性谷氨酸受体5mRNA表达改变及α-甲基-4-羧基苯丙氨酸的治疗作用.中华创伤杂志,2001,17:199-201.]
[16]Wang JW,Wang HD,Cong ZX,Zhang XS,Zhou XM,Zhang DD.Activation of metabotropic glutamate receptor 5 reduces the secondary brain injury after traumatic brain injury in rats.Biochem Biophys Res Commun,2013,430:1016-1021.
[17]Leon R,Wu H,Jin Y,Wei J,Buddhala C,Prentice H,Wu JY.Protective function of taurine in glutamate-induced apoptosis in cultured neurons.J Neurosci Res,2009,87:1185-1194.
[18]Wang JW,Wang HD.The research progress of metabotropic glutamate receptor 5 in neurogliocyte.Zhonghua Shen Jing Wai Ke Ji Bing Yan Jiu Za Zhi,2013,12:470-471.[王嘉炜,王汉东.代谢性谷氨酸受体5在神经胶质细胞中的研究进展.中华神经外科疾病研究杂志,2013,12:470-471.]