微透析法测定脑缺血损伤后黄芩苷、栀子苷及兴奋性氨基酸含量变化
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摘要
目的:建立微透析检测脑缺血损伤后脑脊液中黄芩苷、栀子苷及兴奋性氨基酸含量变化方法,阐明黄芩苷、栀子苷脑保护作用物质基础。方法:采用微透析技术收集给药前后脑缺血大鼠的脑脊液,应用液质联用技术动态监测黄芩苷、栀子苷及兴奋性氨基酸的含量变化规律,对黄芩苷、栀子苷配伍抗脑缺血损伤作用进行研究。结果:黄芩苷与栀子苷配伍(7∶3)给药7天后,微透析技术收集大鼠脑缺血前后海马部位的脑脊液,利用液质联用技术进行检测,仅检测到栀子苷、谷氨酸和天冬氨酸三种物质,同时栀子苷、黄芩苷配伍45mg/kg、60mg/kg组对兴奋性氨基酸的释放起到抑制作用。结论:栀子苷可以透过血脑屏障,推测黄芩苷、栀子苷(7∶3)配伍可能通过抑制兴奋性氨基酸的释放而发挥脑保护作用。
Objective: To establish the method to determine the contents of baicalin( BC),geniposide( GP) and EAAs in cerebrospinal fluid after cerebral ischemic injury by microdialysis,and study the material basis of their protective effects. Methods: The cerebrospinal fluid( CSF) of cerebral ischemia rats was collected by microdialysis technique before and after the administration,the contents of BC,GP and EAAs were dynamically monitored by liquid-mass spectrometry to study the protective effect of BC and GP on cerebral ischemia injury. Results: Mixture of BC and GP( 7: 3) was administered for 7 days,then CSF of rats hippocampus before and after the administration were collected by microdialysis,and only GP,Glu and Asp were detected by HPLC-MS/MS. Release of EAAs was inhibited by 60 mg/kg BC and 45 mg/kg GP group. Conclusion: The result shows that GP can penetrate the blood-brain barrier. In addition,the compatibility of BC and GP( 7: 3) may inhibit the release of EAAs from hippocampal tissue to extracellular fluid,to protect the brain.
Objective: To establish the method to determine the contents of baicalin( BC),geniposide( GP) and EAAs in cerebrospinal fluid after cerebral ischemic injury by microdialysis,and study the material basis of their protective effects. Methods: The cerebrospinal fluid( CSF) of cerebral ischemia rats was collected by microdialysis technique before and after the administration,the contents of BC,GP and EAAs were dynamically monitored by liquid-mass spectrometry to study the protective effect of BC and GP on cerebral ischemia injury. Results: Mixture of BC and GP( 7: 3) was administered for 7 days,then CSF of rats hippocampus before and after the administration were collected by microdialysis,and only GP,Glu and Asp were detected by HPLC-MS/MS. Release of EAAs was inhibited by 60 mg/kg BC and 45 mg/kg GP group. Conclusion: The result shows that GP can penetrate the blood-brain barrier. In addition,the compatibility of BC and GP( 7: 3) may inhibit the release of EAAs from hippocampal tissue to extracellular fluid,to protect the brain.
引文
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[10]朱黎霞,张英丰.基于微透析技术结合液质联用的丹酚酸B正常和高脂血症大鼠药动学比较研究.中草药,2015,46(1)∶90～95.
[11]陈科达,王梦令,葛求富, 等. 黄芩苷、栀子苷及其配伍治疗局灶性脑缺血大鼠药效评价及作用机制研究. 中国药学杂志, 2006, 14∶ 1234～1236.
[12]王斌, 李敏, 侯建平, 等. 栀子、黄芩药材与标准品对小鼠脑缺血保护作用的研究.中药药理与临床, 2010; 26(5)∶ 74～76.
[13]李传云, 潘彦舒, 贾旭, 等. 黄芩、栀子配伍对大鼠局灶性脑缺血再灌注模型缺血级联反应的影响.北京中医药大学学报,2002, 25(6)∶ 31～33.
[14]朱晓磊,张娜,李澎涛,等. 栀子苷阻抑脑缺血损伤级联反应的作用环节探讨.中国药学杂志.2004,29(11)∶1065～1068.
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[18]耿小茵,赖真,邓常青,等.补阳还五汤对鼠脑缺血再灌注后脑组织兴奋性氨基酸的作用.广州中医药大学学报,2004,21(2)∶125～127.
[19]李伟.软脉胶囊对脑缺血再灌注大鼠兴奋性和抑制性氨基酸的影响.中国中医急症, 2001,10(6)∶348～349.
[20]刘向哲.中风星蒌通腑胶囊对急性脑缺血大鼠脑组织中氨基酸平衡的影响.中国中医急症,2004,13(9)∶609～610.
[21]李花,刘旺华,刘建新,等.丹龙醒脑片对沙土鼠脑缺血再灌注保护作用的实验研究.中国中医药信息杂志,2003,10(5)∶29～31.
[22]Zhou H X,Wollmuth L P.Advancing NMDA receptor physiol-ogy by integrating multiple approaches.Trends Neurosci,2017,40∶129～137.
[2]Laird M D, Meiler S E, Vender J R, et al. Curcumin attenuates cerebral edema following traumaticbraininjuryinmice∶apossibleroleforaquaporin-4 Journal of Neurochemistry,2010,113(3)∶637～648.
[3]Stamatovic S M, Keep R F, Andjelkovic A V. Brain Endothelial Cell-Cell Junctions: How to "Open" the Blood Brain Barrier. Current Neuropharmacology,2008,6(3)∶179～192.
[4]Turner R,Vink R. Inhibition of neurogenic inflammation as a novel treatment for ischemic stroke. Timely Topics in Medicine Cardiovascular Diseases,2007,11(4)∶221～226.
[5]程方敏,刘在贵. 脑梗死患者脑脊液兴奋性氨基酸含量的测定及其意义 ,中华现代内科学杂志2008,5(8)∶349～350.
[6]疏树华,方才.兴奋性氨基酸及受体与脑损伤的研究进展. 立体定向和功能性神经外科杂志,2007,20(2)∶124～126.
[7]李宁,吴建中,蒋京生.脑缺血时海马细胞外液中兴奋性氨基酸的变化.中国病理生理杂志,1993,9(6)∶740～744.
[8]李菊,杨期东,欧阳珊,等.脑缺血大鼠海马细胞外液氨基酸的动态变化.中华医学杂志,1996,76(4)∶294～296.
[9]武祺, 胡亮,黄惠敏. 50例急性大脑皮层梗死患者脑脊液兴奋性氨基酸含量的研究.求医问药:学术版,2011,9(12)∶217～218.
[10]朱黎霞,张英丰.基于微透析技术结合液质联用的丹酚酸B正常和高脂血症大鼠药动学比较研究.中草药,2015,46(1)∶90～95.
[11]陈科达,王梦令,葛求富, 等. 黄芩苷、栀子苷及其配伍治疗局灶性脑缺血大鼠药效评价及作用机制研究. 中国药学杂志, 2006, 14∶ 1234～1236.
[12]王斌, 李敏, 侯建平, 等. 栀子、黄芩药材与标准品对小鼠脑缺血保护作用的研究.中药药理与临床, 2010; 26(5)∶ 74～76.
[13]李传云, 潘彦舒, 贾旭, 等. 黄芩、栀子配伍对大鼠局灶性脑缺血再灌注模型缺血级联反应的影响.北京中医药大学学报,2002, 25(6)∶ 31～33.
[14]朱晓磊,张娜,李澎涛,等. 栀子苷阻抑脑缺血损伤级联反应的作用环节探讨.中国药学杂志.2004,29(11)∶1065～1068.
[15]Arranz A M, Gottlieb M, Pérezcerdá F, et al. Increased expression of glutamate transporters in subcortical white matter after transient focal cerebral ischemia. Neurobiology of Disease, 2010, 37(1)∶156～165.
[16]Lipton S A, Rosenberg P A. Excitatory amino acids as a final common pathway for neurologic disorders.. N Engl J Med, 1994, 330(9)∶613～622.
[17]Cazeviedle C,Muller A,Meynier F,et al. Superoxideandnotrieoxidecooperationinhypoxa/reoxygenution-inducedneuroninjury. Free Radle Biol Med,1993,14(4)∶ 389～395.
[18]耿小茵,赖真,邓常青,等.补阳还五汤对鼠脑缺血再灌注后脑组织兴奋性氨基酸的作用.广州中医药大学学报,2004,21(2)∶125～127.
[19]李伟.软脉胶囊对脑缺血再灌注大鼠兴奋性和抑制性氨基酸的影响.中国中医急症, 2001,10(6)∶348～349.
[20]刘向哲.中风星蒌通腑胶囊对急性脑缺血大鼠脑组织中氨基酸平衡的影响.中国中医急症,2004,13(9)∶609～610.
[21]李花,刘旺华,刘建新,等.丹龙醒脑片对沙土鼠脑缺血再灌注保护作用的实验研究.中国中医药信息杂志,2003,10(5)∶29～31.
[22]Zhou H X,Wollmuth L P.Advancing NMDA receptor physiol-ogy by integrating multiple approaches.Trends Neurosci,2017,40∶129～137.