芳香开窍类中药对星形胶质细胞水通道蛋白4表达的影响
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摘要
摘要:水通道蛋白-4(aquaporins-4,AQP4)是中枢神经系统的水通道,它主要分布在血-脑,脑-脑脊液界面,意味着AQP4在正常和病理生理情况下维持脑内水平衡起重要作用,本文介绍了AQP4在脑内的机构,分布、功能及其在脑水肿的形成及清除中的作用及其可能的调节机制。据此,研究各种药物调节AQP4表达的药物对创伤,脑肿瘤,癫痫及其他原因导致脑水稳态失衡而引起的脑功能障碍有很大作用。
目的:通过研究家兔含药脑脊液对原代培养的小鼠大脑星形胶质细胞水通道基因表达的影响,探讨芳香开窍类中药在治疗中枢神经疾病时的作用机理。方法:给家兔灌服麝香溶液、石菖蒲挥发油溶液,40min后抽取脑脊液,用含药脑脊液刺激原代培养的小鼠大脑星形胶质细胞,RT-PCR方法检测星形胶质细胞水通道-4基因表达水平。结果:原代培养的小鼠大脑星形胶质细胞表达水通道-4mRNA,培养液中加入家兔脑脊液对水通道-4mRNA表达没有影响,含石菖蒲的含药脑脊液使原代培养的小鼠大脑星形胶质细胞水通道-4基因表达降低;而含麝香的含药脑脊液对原代培养的小鼠大脑星形胶质细胞水通道-4基因表达无明显影响。结论:芳香开窍药石菖蒲挥发油的神经药理作用之一可能是降低星形胶质细胞AQP4基因的表达。
The AQPs are a family of homologous water channel proteins(number at least 1 3 in mammals that provide the major route for water movement across plasma membranes in a variety of cell types and play a key role in maintain the water balance in brain and the microenvironment in cell.
Aquaporin-4(AQP4)is the major water channel in the CNS , Its expression at fluid-tissue barrier (blood-brain and brain-cerebrospinal fluid barriers)throughout the brain and spinal cord suggests a role in water transport under normal and pathological conditions AQP4.Further insight into the mechanisms by which brain AQPs are regulated will be of utmost clinical importance,since perturbed water flow via brain AQPs has been implicated in many neurological disease。
Objective To study the mechanism concerning herbs of resuscitation with aromatics on treating central nerve disease. We investigated the effect of drug-containing cerebrospinal fluid which extracted from rabbits on the expression of aquaporin(AQP) mRNA in primary cultured brain astrocytes (AST) of mice.Methods We stimulate primary cultured brain AST with cerebrospinal fluid drawning out from rabbits treated by intragastric administration of musk solution, solution of volatile oil from acorus tatarinowii, then examine the expression level of AQP4 mRNA by RT-PCR. Results AQP4 mRNA was expressed in the primary cultured brain AST. There was no influence on the expression of AQP4 mRNA when added control cerebrospinal fluid to culture medium. The cerebrospinal fluid contained calamus depress the expression of AQP4 mRNA. However the cerebrospinal fluid contained musk have no remarkable effect on the expression of AQP4 mRNA. Conclusion Valitile oil from acorus tatarinowii may decrease the expression of AQP4 mRNA .
目的:通过研究家兔含药脑脊液对原代培养的小鼠大脑星形胶质细胞水通道基因表达的影响,探讨芳香开窍类中药在治疗中枢神经疾病时的作用机理。方法:给家兔灌服麝香溶液、石菖蒲挥发油溶液,40min后抽取脑脊液,用含药脑脊液刺激原代培养的小鼠大脑星形胶质细胞,RT-PCR方法检测星形胶质细胞水通道-4基因表达水平。结果:原代培养的小鼠大脑星形胶质细胞表达水通道-4mRNA,培养液中加入家兔脑脊液对水通道-4mRNA表达没有影响,含石菖蒲的含药脑脊液使原代培养的小鼠大脑星形胶质细胞水通道-4基因表达降低;而含麝香的含药脑脊液对原代培养的小鼠大脑星形胶质细胞水通道-4基因表达无明显影响。结论:芳香开窍药石菖蒲挥发油的神经药理作用之一可能是降低星形胶质细胞AQP4基因的表达。
The AQPs are a family of homologous water channel proteins(number at least 1 3 in mammals that provide the major route for water movement across plasma membranes in a variety of cell types and play a key role in maintain the water balance in brain and the microenvironment in cell.
Aquaporin-4(AQP4)is the major water channel in the CNS , Its expression at fluid-tissue barrier (blood-brain and brain-cerebrospinal fluid barriers)throughout the brain and spinal cord suggests a role in water transport under normal and pathological conditions AQP4.Further insight into the mechanisms by which brain AQPs are regulated will be of utmost clinical importance,since perturbed water flow via brain AQPs has been implicated in many neurological disease。
Objective To study the mechanism concerning herbs of resuscitation with aromatics on treating central nerve disease. We investigated the effect of drug-containing cerebrospinal fluid which extracted from rabbits on the expression of aquaporin(AQP) mRNA in primary cultured brain astrocytes (AST) of mice.Methods We stimulate primary cultured brain AST with cerebrospinal fluid drawning out from rabbits treated by intragastric administration of musk solution, solution of volatile oil from acorus tatarinowii, then examine the expression level of AQP4 mRNA by RT-PCR. Results AQP4 mRNA was expressed in the primary cultured brain AST. There was no influence on the expression of AQP4 mRNA when added control cerebrospinal fluid to culture medium. The cerebrospinal fluid contained calamus depress the expression of AQP4 mRNA. However the cerebrospinal fluid contained musk have no remarkable effect on the expression of AQP4 mRNA. Conclusion Valitile oil from acorus tatarinowii may decrease the expression of AQP4 mRNA .
引文
[1]杨志军,魏玲星形胶质细胞生物学功能研究进展[J]。中华神经医学杂志,2003, (2):396-398。
[2]才丽平,赵金茹,林庶茹,等。水通道蛋白研究进展[J]。解剖科学进展,2003,9(2):167-170
[3]沈映军。中药药理学。人民卫生出版社。2000。
[4] Solenov E, Watanabe H, Manley G, et al. Sevenfold-reduced osmotic water permeability in primary astrocyte cultures from AQP-4-deficient mice, measured by a fluorescence quenching method[J]. Am J Physiol Cell Physiol. 2004; 286: C426–C432.
[5] Nicchia GP, Frigeri A, Liuzzi GM, et al. Inhibition of AQP4 expression in astrocytes by RNAi determines alterations in cell morphology, growth, and water transport and induces changes in ischemia related genes. The FASEB . 2003, 10:1096-1183.
[6] Binder DK,Yno X,Sick TJ,et a1.Increased seizure duration and slowed potassium kinetics in mice lacking aquaporin-4 water channels.Gila.2006;53:631-636
[7] TS Lee, T Eid, S Mane,et al. Aquaporin-4 is increased in the sclerotic hippocampus in human temporal lobe epilepsy. Acta Neuropathol, 2004; 108(6): 493-502.
[8] Ying-hui Chen,Yong-bo Zhao,Wen-wen Hn,et al. Upregulation of AQP4 expression in the brain of patients with medically intractable epilepsy. Neuroscience Bulletin. 2005; 21(6):413-417
[9]李树蕾,杨立彬,黄艳智,等。中药石菖蒲及其主要成分α-细辛醚对癫痫幼鼠惊厥行为及脑电图的影响。吉林大学学报(医学版)。2006;32(1):74-77
[10]陈俐,廖卫平。石菖蒲萃取挥发油抗癫痫药效研究。中药新药与临床药理。2003;14(3): 171-173
[11]方永奇,魏刚,柯雪红。GC-MS分析石菖蒲挥发油透大鼠血脑屏障的成分研究。中药新药与临床药理。2002;13(3期)181-182
[12]方永奇李翎吴启端。β-细辛醚对小鼠脑组织基因表达谱的影响。中药材。(26)9: 650-652
[13]刘亚敏,张赐安。全脑缺血再灌注大鼠脑组织水通道蛋白-4mRNA的表达及药物对其影响。中国中医急症, 2002;11(6):474-476
[1] Ke C, PoonWS, Ng HK, et al. Heterogeneous responses of aquaporin- 4 in edema formation in a pep licated severe traumatic brain injurymodel in rats [ J ].Neurosci lett, 2001, 23, 301 (1) : 21 - 24.
[2] Saadoun S, PapadopoulosMC, Davies DC, et al. Aquaporin - 4 ex2p ression is increased in edema human brain tumours [ J ]. J NeurolNeu2rosurg Psychiatry, 2002, 72 (2) : 262 - 265.
[3] Davies DC. Blood - brain barrier breakdown in sep tic encephalopathy and brain tumours [ J ]. J Anat, 2002, 200 (6) : 639– 646l32]Binder DK,O~1.1lO K,Ma T,et a1.Increased seizure threshold in mice lacking aquaporin-4 water channels[J].Neuroropoa,2OO4,15:259-262
[4]Manley GT,Fujimma M,Ma T,et a1.Aqusporin-4 deletion in mice reduces brainedema after aeu|e'l~erintoxication andischemie st~ke[J] .Nat Med .2OOO。6:159—163
[5]石向群,杨金升,包仕尧,等.AQP4在正常大鼠脑内的分布[J].中华神经医学杂志,2004,13:257.260
[6]包仕尧,石向群,张志琳,等.大鼠脑缺血再灌注损伤过程中水通道,蛋白4表达水平的变化[J].国外医学.脑血管疾病分册, 2OO4,12:122-126
[7]李燕华,孙善全.水通道蛋白一4在脑出血大鼠脑组织的分布.临床神经病学杂志,2004,17(2):108—110.
[8]李燕华,孙善全.低渗液对星形胶质细胞水通道蛋白一4表达的影响.中华医学杂志,2004,8(6):496—501.
[9]鲁宏,孙善全.缺血性脑水肿的AQP4表达与磁共振成像的相关性研究.中国医学影像杂志,2003,19(8):957—960.
[10]Yamamoto N ,Yoneda K,Asai K,et a1.Alterations in the expression of the AQP family in cultured rat astrocytes during hypoxia and reoxygenation.Brain Res Mol Brain Res,2001,90:26.
[11]Preston GM ,Agre P.Isolation of the cDNA for erythrocyte integral membrane protein of 28 kilodahons:member of an ancient channel family.Proc Natl Acad Sci USA ,1991,88:11110—11114.
[12]陈彗星,顾承鳞.水通道蛋白一4研究进展.中国基层医药,2004,11(12):1521—1522.
[13] Umenishi F,Verkman AS,Gropper MA.Quantitative analysis of aquaporin mRNA expression in rat tissues by Rnase protection assay.DNA Cell Biol,1996,15(6):475—478.
[14] Zelenin S,Gunnarson E,Allklna T,et a1.Identification of a new form of AQP4 mRNA that is develnpmentally expressed in mouse brain. Pediatr—Res.2000,48(3):335—339.
[15] Neely JD,Nielsen S,Agre P,et a1.Heterotetramerlc composition of aquaporin-4 water channels.Biochemistry,1999,38(34):1 1 156
[16]Rash JE,Yasumara T,hudson CS,et a1.Direct immunogold labeling of aquaporin-4 square arrays of astrocyte and ependymocyte plasma membranes in rat brain and spinal cord.Pioc Natl Acad Sci USA,1998,95(20):11981—11985.
[17]李燕华,孙善全.水通道蛋白一4在脑出血大鼠脑组织的分布.临床神经病学杂志,2004,17(2):108—110.
[18] Yamamoto N ,Yoneda K,Asai K,et a1.Alterations in the expression of the AQP family in cultured rat astrocytes during hypoxia and reoxygenation.Brain Res Mol Brain Res,2001,90:26.
[19]李燕华,孙善全.低渗液对星形胶质细胞水通道蛋白一4表达的影响.中华医学杂志,2004,8(6):496—501.
[20]鲁宏,孙善全.缺血性脑水肿的AQP4表达与磁共振成像的相关性研究.中国医学影像杂志,2003,19(8):957—960
[2]才丽平,赵金茹,林庶茹,等。水通道蛋白研究进展[J]。解剖科学进展,2003,9(2):167-170
[3]沈映军。中药药理学。人民卫生出版社。2000。
[4] Solenov E, Watanabe H, Manley G, et al. Sevenfold-reduced osmotic water permeability in primary astrocyte cultures from AQP-4-deficient mice, measured by a fluorescence quenching method[J]. Am J Physiol Cell Physiol. 2004; 286: C426–C432.
[5] Nicchia GP, Frigeri A, Liuzzi GM, et al. Inhibition of AQP4 expression in astrocytes by RNAi determines alterations in cell morphology, growth, and water transport and induces changes in ischemia related genes. The FASEB . 2003, 10:1096-1183.
[6] Binder DK,Yno X,Sick TJ,et a1.Increased seizure duration and slowed potassium kinetics in mice lacking aquaporin-4 water channels.Gila.2006;53:631-636
[7] TS Lee, T Eid, S Mane,et al. Aquaporin-4 is increased in the sclerotic hippocampus in human temporal lobe epilepsy. Acta Neuropathol, 2004; 108(6): 493-502.
[8] Ying-hui Chen,Yong-bo Zhao,Wen-wen Hn,et al. Upregulation of AQP4 expression in the brain of patients with medically intractable epilepsy. Neuroscience Bulletin. 2005; 21(6):413-417
[9]李树蕾,杨立彬,黄艳智,等。中药石菖蒲及其主要成分α-细辛醚对癫痫幼鼠惊厥行为及脑电图的影响。吉林大学学报(医学版)。2006;32(1):74-77
[10]陈俐,廖卫平。石菖蒲萃取挥发油抗癫痫药效研究。中药新药与临床药理。2003;14(3): 171-173
[11]方永奇,魏刚,柯雪红。GC-MS分析石菖蒲挥发油透大鼠血脑屏障的成分研究。中药新药与临床药理。2002;13(3期)181-182
[12]方永奇李翎吴启端。β-细辛醚对小鼠脑组织基因表达谱的影响。中药材。(26)9: 650-652
[13]刘亚敏,张赐安。全脑缺血再灌注大鼠脑组织水通道蛋白-4mRNA的表达及药物对其影响。中国中医急症, 2002;11(6):474-476
[1] Ke C, PoonWS, Ng HK, et al. Heterogeneous responses of aquaporin- 4 in edema formation in a pep licated severe traumatic brain injurymodel in rats [ J ].Neurosci lett, 2001, 23, 301 (1) : 21 - 24.
[2] Saadoun S, PapadopoulosMC, Davies DC, et al. Aquaporin - 4 ex2p ression is increased in edema human brain tumours [ J ]. J NeurolNeu2rosurg Psychiatry, 2002, 72 (2) : 262 - 265.
[3] Davies DC. Blood - brain barrier breakdown in sep tic encephalopathy and brain tumours [ J ]. J Anat, 2002, 200 (6) : 639– 646l32]Binder DK,O~1.1lO K,Ma T,et a1.Increased seizure threshold in mice lacking aquaporin-4 water channels[J].Neuroropoa,2OO4,15:259-262
[4]Manley GT,Fujimma M,Ma T,et a1.Aqusporin-4 deletion in mice reduces brainedema after aeu|e'l~erintoxication andischemie st~ke[J] .Nat Med .2OOO。6:159—163
[5]石向群,杨金升,包仕尧,等.AQP4在正常大鼠脑内的分布[J].中华神经医学杂志,2004,13:257.260
[6]包仕尧,石向群,张志琳,等.大鼠脑缺血再灌注损伤过程中水通道,蛋白4表达水平的变化[J].国外医学.脑血管疾病分册, 2OO4,12:122-126
[7]李燕华,孙善全.水通道蛋白一4在脑出血大鼠脑组织的分布.临床神经病学杂志,2004,17(2):108—110.
[8]李燕华,孙善全.低渗液对星形胶质细胞水通道蛋白一4表达的影响.中华医学杂志,2004,8(6):496—501.
[9]鲁宏,孙善全.缺血性脑水肿的AQP4表达与磁共振成像的相关性研究.中国医学影像杂志,2003,19(8):957—960.
[10]Yamamoto N ,Yoneda K,Asai K,et a1.Alterations in the expression of the AQP family in cultured rat astrocytes during hypoxia and reoxygenation.Brain Res Mol Brain Res,2001,90:26.
[11]Preston GM ,Agre P.Isolation of the cDNA for erythrocyte integral membrane protein of 28 kilodahons:member of an ancient channel family.Proc Natl Acad Sci USA ,1991,88:11110—11114.
[12]陈彗星,顾承鳞.水通道蛋白一4研究进展.中国基层医药,2004,11(12):1521—1522.
[13] Umenishi F,Verkman AS,Gropper MA.Quantitative analysis of aquaporin mRNA expression in rat tissues by Rnase protection assay.DNA Cell Biol,1996,15(6):475—478.
[14] Zelenin S,Gunnarson E,Allklna T,et a1.Identification of a new form of AQP4 mRNA that is develnpmentally expressed in mouse brain. Pediatr—Res.2000,48(3):335—339.
[15] Neely JD,Nielsen S,Agre P,et a1.Heterotetramerlc composition of aquaporin-4 water channels.Biochemistry,1999,38(34):1 1 156
[16]Rash JE,Yasumara T,hudson CS,et a1.Direct immunogold labeling of aquaporin-4 square arrays of astrocyte and ependymocyte plasma membranes in rat brain and spinal cord.Pioc Natl Acad Sci USA,1998,95(20):11981—11985.
[17]李燕华,孙善全.水通道蛋白一4在脑出血大鼠脑组织的分布.临床神经病学杂志,2004,17(2):108—110.
[18] Yamamoto N ,Yoneda K,Asai K,et a1.Alterations in the expression of the AQP family in cultured rat astrocytes during hypoxia and reoxygenation.Brain Res Mol Brain Res,2001,90:26.
[19]李燕华,孙善全.低渗液对星形胶质细胞水通道蛋白一4表达的影响.中华医学杂志,2004,8(6):496—501.
[20]鲁宏,孙善全.缺血性脑水肿的AQP4表达与磁共振成像的相关性研究.中国医学影像杂志,2003,19(8):957—960
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