逍遥散对慢性应激损伤模型大鼠海马区Cacnb4基因表达的影响
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摘要
目的:本研究用逍遥散干预慢性应激损伤模型大鼠,观察其海马区Cacnb4基因的表达情况。方法:以60只Wistar大鼠随机分为空白对照组、慢性应激模型组、逍遥散治疗组3组,每组20只。造模方法采用Cart多相性慢性应激大鼠模型并加以改进,逍遥散治疗组在造模同时给予逍遥散2m L/(只·d),连续造模21 d,于第22天处死大鼠并取材。采用RT-PCR一步法及免疫组化检测方法测定各组大鼠海马区域Cacnb4基因及其蛋白表达情况。结果:慢性应激模型组大鼠海马Cacnb4mRNA及其相应蛋白表达均显著上调,中药复方逍遥散可从基因及蛋白水平同时纠正这种上调的表达变化。结论:电压依赖型钙离子通道β4辅助亚基的表达变化参与了慢性应激损伤过程,逍遥散对慢性应激所致神经细胞损伤可能通过调节这一基因发挥保护性作用。
Objective: We treat the chronic stress model by Xiaoyao Powder,observing the expression of Cacnb4 gene in brain hippocampus. Methods: Sixty Wistar rats were selected and randomly divided into three groups,blank group,model group and treatment group of Xiaoyao Powder,20 rats in each group. The modeling method was put to use Cart heterogenicity chronic stress rat model and improved to establish the rat model fou 21 days and drugs including Xiaoyao Powder 2 m L to each rat every day for treatment group of Xiaoyao Powder. On the 22 nd day,we observed the expression of Cacnb4 gene and protein in hippocampus of each group rats by Real time-PCR and immunohistochemistry method. Results: The expressions of Cacnb4 mRNA and its corresponding protein in hippocampus of chronic stress model group were significantly increased. Xiaoyao Powder can also correct the expression changes of this up regulation from the level of gene and protein. Conclusion: Voltage-dependent Calcium Ion Channelβ4 Secondary Subunit expression changes involved in the process of chronic stress injury. Xiaoyao Powder on chronic stress-induced nerve cell damage may play a protective regulatory role through this gene.
Objective: We treat the chronic stress model by Xiaoyao Powder,observing the expression of Cacnb4 gene in brain hippocampus. Methods: Sixty Wistar rats were selected and randomly divided into three groups,blank group,model group and treatment group of Xiaoyao Powder,20 rats in each group. The modeling method was put to use Cart heterogenicity chronic stress rat model and improved to establish the rat model fou 21 days and drugs including Xiaoyao Powder 2 m L to each rat every day for treatment group of Xiaoyao Powder. On the 22 nd day,we observed the expression of Cacnb4 gene and protein in hippocampus of each group rats by Real time-PCR and immunohistochemistry method. Results: The expressions of Cacnb4 mRNA and its corresponding protein in hippocampus of chronic stress model group were significantly increased. Xiaoyao Powder can also correct the expression changes of this up regulation from the level of gene and protein. Conclusion: Voltage-dependent Calcium Ion Channelβ4 Secondary Subunit expression changes involved in the process of chronic stress injury. Xiaoyao Powder on chronic stress-induced nerve cell damage may play a protective regulatory role through this gene.
引文
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[11]杨胜,刘振伟,万勤,等.大鼠海马神经元膜离子通道随培养时间变化的特点[J].中国应用生理学杂志,2004(2):151-155.
[12]Etemad S,Campiglio M,Obermair GJ,et al.The juvenile myoclonic epilepsy mutant of the calcium channelβ4 subunit displays normal nuclear targeting in nerve and muscle cells[J].Channels(Austin),2014,8(4):334-343.
[13]Subramanyam P,Obermair GJ,Baumgartner S,et al.Activity and calcium regulate nuclear targeting of the calcium channel beta4b subunit in nerve and muscle cells[J].Channels(Austin),2009,3(5):343-355.
[14]Etemad S,Obermair GJ,Bindreither D,et al.Differential neuronal targeting of a new and two known calcium channelβ4 subunit splice variants correlates with their regulation of gene expression[J].J Neurosic,2014,34(4):1446-1461.
[15]Ronjat M,Kiyonaka S,Barbado M,et al.Nuclear life of the voltagegated Cacnb4 subunit and its role in gene transcription regulation[J].Channel(Austin),2013,7(2):119-125.
[16]Tadmouri A,Kiyonaka S,Barbado M,et al.Cacnb4 directly couples electrical activity to gene expression,a process defective in juvenile epilepsy[J].EMBO J,2012,31(18):3730-3744.
[2]任双,吴高峰,胡建民.慢性应激与大脑海马神经再生的关系[J].动物医学进展,2016(1):85-88.
[3]敖海清,徐志伟,严灿,等.逍遥散对应激大鼠海马突触体内PKC活性及Ca2+浓度的影响[J].山东中医杂志,2006,(2):112-114.
[4]崔越,周敏,周静文,等.海马神经元钙离子通道的研究进展[J].中国老年学杂志,2012(3):643-645.
[5]Solmaz Etemad,Marta Campiglio,Gerald J Obermair,et al.The juvenile myoclonic epilepsy mutant of the calcium channelβ4 subunit displays normal nuclear targeting in nerve and muscle cells[J].Channels(Austin),2014,8(4):334-343.
[6]Zamponi GW,Striessnig J,Koschak A,et al.The Physiology,pathology,and pharmacology of voltage-gated calcium channels and their future therapeutic potential[J].Pharmacological Reviews,2015,67(4):821-870.
[7]Subramanyam P,Obermair GJ,Baumgartner S,et al.Activity and calcium regulate nuclear targeting of the calcium channel beta4b subunit in nerve and muscle cells[J].Channels,2009,3(5):343-355.
[8]Schjtt JM,Hsu SC,Plummer MR.The neuronal beta 4 subunit increases the unitary conductance of L-type voltage-gated calcium channels in PC12 cells[J].Journal of Biological Chemistry,2003,278(36):33936-33942.
[9]Bernard F,Ramirez DS,Sun S,et al.L-type calcium channel antagonism-Translation from in vitro to in vivo[J].Journal of Pharmacological and Toxicological Methods,2017:86-92.
[10]QU Yu.Changes in Single L-Type Calcium Channel Currents in CA1 Pyramidal Neurons of Rat Hippocampus After Transient Forebrain Ischemia[J].Progress in Biochemistry and Biophysics,2003,30(5):755-760.
[11]杨胜,刘振伟,万勤,等.大鼠海马神经元膜离子通道随培养时间变化的特点[J].中国应用生理学杂志,2004(2):151-155.
[12]Etemad S,Campiglio M,Obermair GJ,et al.The juvenile myoclonic epilepsy mutant of the calcium channelβ4 subunit displays normal nuclear targeting in nerve and muscle cells[J].Channels(Austin),2014,8(4):334-343.
[13]Subramanyam P,Obermair GJ,Baumgartner S,et al.Activity and calcium regulate nuclear targeting of the calcium channel beta4b subunit in nerve and muscle cells[J].Channels(Austin),2009,3(5):343-355.
[14]Etemad S,Obermair GJ,Bindreither D,et al.Differential neuronal targeting of a new and two known calcium channelβ4 subunit splice variants correlates with their regulation of gene expression[J].J Neurosic,2014,34(4):1446-1461.
[15]Ronjat M,Kiyonaka S,Barbado M,et al.Nuclear life of the voltagegated Cacnb4 subunit and its role in gene transcription regulation[J].Channel(Austin),2013,7(2):119-125.
[16]Tadmouri A,Kiyonaka S,Barbado M,et al.Cacnb4 directly couples electrical activity to gene expression,a process defective in juvenile epilepsy[J].EMBO J,2012,31(18):3730-3744.