Effect of Modified Xiao Chaihutang on Expression of EAATs and VGLUTs in Hippocampus of Chronic Restraibt Stress Depression Model Rat

Er-ping XU; Meng-di MAO; Li-zhi SHANG; Guo-qiang WANG; Bao-guang LIU; Yao-yang LI; Ming BA

doi:10.13422/j.cnki.syfjx.20202202

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Effect of Modified Xiao Chaihutang on Expression of EAATs and VGLUTs in Hippocampus of Chronic Restraibt Stress Depression Model Rat

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- Effect of Modified Xiao Chaihutang on Expression of EAATs and VGLUTs in Hippocampus of Chronic Restraibt Stress Depression Model Rat
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 23, Pages: 27-36(2020)
- 作者机构：
  
  1.河南中医药大学中医药科学院，郑州 450046
  2.河南省仲景方药现代研究重点实验室，郑州 450046
  3.河南中医药大学基础医学院，郑州 450046
  4.河南中医药大学第一临床医学院，郑州 450006
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20202202
  CLC： R2-0;R22;R285.5;R289
- Received：21 May 2020，
  
  Published Online：17 September 2020，
  
  Published：05 December 2020
- 稿件说明：
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许二平,毛梦迪,尚立芝等.小柴胡汤加味对慢性束缚抑郁模型大鼠海马谷氨酸转运体EAATs，VGLUTs表达的影响[J].中国实验方剂学杂志,2020,26(23):27-36.

XU Er-ping,MAO Meng-di,SHANG Li-zhi,et al.Effect of Modified Xiao Chaihutang on Expression of EAATs and VGLUTs in Hippocampus of Chronic Restraibt Stress Depression Model Rat[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(23):27-36.
许二平,毛梦迪,尚立芝等.小柴胡汤加味对慢性束缚抑郁模型大鼠海马谷氨酸转运体EAATs，VGLUTs表达的影响[J].中国实验方剂学杂志,2020,26(23):27-36. DOI： 10.13422/j.cnki.syfjx.20202202.

XU Er-ping,MAO Meng-di,SHANG Li-zhi,et al.Effect of Modified Xiao Chaihutang on Expression of EAATs and VGLUTs in Hippocampus of Chronic Restraibt Stress Depression Model Rat[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(23):27-36. DOI： 10.13422/j.cnki.syfjx.20202202.

摘要

目的

观察小柴胡汤加味对慢性束缚抑郁模型大鼠海马谷氨酸膜转运体（EAATs）及囊泡转运体（VGLUTs）表达的影响，探讨小柴胡汤加味基于谷氨酸转运的抗抑郁机制。

方法

120只SD大鼠随机分为正常组、模型组、小柴胡汤加味组低、中、高剂量组、利鲁唑组，每组20只。除正常组外，其余各组采用束缚应激制备大鼠抑郁模型，小柴胡汤加味低、中、高剂量组分别灌胃（

）小柴胡汤加味6.5，13，26 g·kg

-1

；利鲁唑组腹腔注射利鲁唑20 mg·kg

-1

；正常组和模型组

等量生理盐水；1次/d，共干预21 d。采用强迫游泳实验（FST）和悬尾实验（TST）评价大鼠的抑郁行为；采用高效液相色谱法（HPLC）检测海马组织中谷氨酸含量；实时荧光定量聚合酶链式反应（Real-time PCR）检测大鼠海马中EAAT1，EAAT2和EAAT3 mRNA表达；蛋白免疫印迹法（Western blot）检测海马组织EAAT1，EAAT2，EAAT3，VGLUT1，VGLUT2蛋白表达；用尼氏染色法观察大鼠海马神经元形态，免疫组化（IHC）检测海马CA1区EAAT1，EAAT2和NeuN蛋白在神经细胞中的定位表达。

结果

与正常组比较，模型组大鼠悬尾和强迫游泳不动时间均显著延长（

0.01），海马内EAAT1，EAAT2，EAAT3 mRNA和蛋白表达均显著下降（

0.01），VGLUT1和NeuN蛋白表达均显著降低（

0.01）；而谷氨酸水平和VGLUT2表达均显著增高（

0.01）。与模型组比较，小柴胡汤加味中、高剂量组大鼠悬尾和强迫游泳不动时间明显缩短（

0.05，

0.01），海马内EAAT1，EAAT2和EAAT3 mRNA和蛋白表达量均显著增加（

0.01），VGLUT1和NeuN蛋白表达均显著增强（

0.01），谷氨酸水平和VGLUT2表达显著回降（

0.01），海马神经元结构明显复原。

结论

小柴胡汤加味有明显的抗抑郁作用，其机制可能与其上调大鼠海马内EAAT1，EAAT2，EAAT3基因和VGLUT1蛋白表达。

Abstract

Objective

To study the effect of modified Xiao Chaihutang on the expressions of excitatory amino acid transporters（EAATs） and vesicle glutamate transporters（VGLUTs）in hippocampus of rats with chronic depression， in order to explore the anti-depressant mechanism of modified Xiao Chaihutang based on glutamate transport.

Method

A total of 120 SD rats were randomly divided into normal group， model group， and low， middle and high-dose modified Xiaochaihutang groups （6.5， 13， 26 g·kg

-1

） and riluzole group， with 20 rats in each group．Except normal group， the depression model of rats was prepared through Chronic restraint stress（CRS）. The normal group and the model group were intragastrically （

） given normal saline. The modified Xiao Chaihutang groups were intragastrically given corresponding herbal drugs （6.5， 13， 26 g·kg

-1

）， and the Riluzole group was given Riluzole 20 mg·kg

-1

through intraoeritoneal injection for 21 days， once a day. Then the depressive behaviors of rats were observed by forced swimming test （FST） and tail suspension test （TST）. The level of glutamic acid （Glu） in rats hippocampus was determined by high performance liquid chromatography （HPLC）. The mRNA expressions of EAAT1， EAAT2 and EAAT3 in hippocampus were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）method. Western blot was used to detect the protein expressions of EAAT1， EAAT2， EAAT3， VGLUT1 and VGLUT2 in rat hippocampus tissue. Nissl staining was used to observe the morphology of hippocampal neurons in rats. Immunohistochemical（IHC）S-P method were used to detect the location expressions of EAAT1， EAAT2 and NeuN proteins in rat hippocampal CA1 region tissue.

Result

The immobility times in FST and TST were increased significantly（

0.01）， the mRNA and protein expressions of EAAT1，EAAT2，EAAT3 were decreased significantly

（

0.01）， and as well as the expressions of VGLUT1 and NeuN were decreased significantly（

0.01）， while the level of Glutamate and the expression of VGLUT2 were increased significantly（

0.01） in model group， compared with normal group. Compared with model group，the immobility times in FST and TST were decreased significantly（

0.05，

0.01）， mRNA and protein expressions of EAAT1，EAAT2，EAAT3 were increased significantly（

0.01）， and expressions of VGLUT1 and NeuN were increased significantly（

0.01）. However， the level of Glutamate and the expression of VGLUT2 were decreased significantly（

0.01）， and the damage of hippocampal neurons in rats was mild in middle and high-dose modified Xiao Chaihutang groups.

Conclusion

Modified Xiao Chaihutang has an anti-depressive effect. Its mechanism may be related to its up-regulation of expressions of EAAT1， EAAT2， EAAT3 genes and VGLUT1 protein in the hippocampus of depression model rats.

关键词

Keywords

references

FANG X ， ZHANG C ， WU Z ， et al . The association between somatic symptoms and suicidal ideation in Chinese ﬁrst-episode major depressive disorder ［J］. J Affect Disord ， 2019 ， 245 （ 2 ）： 17 - 21

RUSH A J ， TRIVEDI M H ， WISNIEWSKI S R ， et al . Acute and longer-term outcomes in depressed out patients requiring one or several treatment steps： a STAR-D report ［J］. Am J Psychiatry ， 2006 ， 163 （ 11 ）： 1905 - 1917 .

LINDSTROM ， SARAH H ， SUNDBERG ， et al . VGluT1 deficiency impairs visual attention and reduces the dynamic range of short-term plasticity at corticothalamic synapses ［J］. Cereb Cortex ， 2020 ， 30 （ 3 ）： 1813 - 1829 .

SHEN ， ZHIJUN ， HARAGOPAL ， et al . Zinc modulates synaptic transmission by differentially regulating synaptic glutamate homeostasis in hippocampus ［J］. Eur J Neurosci ， 2020 ， doi： 10.1111/ejn.14749 http://dx.doi.org/10.1111/ejn.14749 .

杜林柯，王萌，周永学 . 《金匮要略》从脏腑论治神志病学术思想探析［J］. 中医学报， 2020 ， 35 （ 2 ）： 257 - 259 .

刘丽军，刘思尧，沙春河，等 . 柴越汤对抑郁症大鼠 HPA 轴的调节作用［J］. 中国实验方剂学杂志， 2017 ， 23 （ 10 ）： 133 - 138 .

孙永康，杨海燕，王新志 . 王新志分期论治郁证经验［J］. 中国中医基础医学杂志， 2020 ， 26 （ 1 ）： 132 - 134 .

MELIA K R ， RYABININAE SCHROEDERR ， et al . Induction and habituation of immediate early gene expression in rat brain by acute and repeated restraint stress ［J］. J Neurosci ， 1994 ， 14 （ 10 ）： 5929 - 5938 .

BRENNAN B P ， HUDSON J I ， JENSEN J E ， et al . Rapid enhancement of glutamatergic neurotransmission in bipolar depression following treatment with riluzole ［J］. Neuropsychopharmacology ， 2010 ， 35 （ 3 ）： 834 - 846 .

戴若以，顾小红，张云东，等 . 束缚应激大鼠下丘脑谷氨酸受体变化和行为绝望状态的关系研究［J］. 重庆医科大学学报， 2016 ， 41 （ 1 ）： 38 - 43 .

KANEKAR S ， BOGDANOVA O V ， OLSON P R ， et al . Hypobaric hypoxia induces depression-like behavior in female sprague-dawley rats，butnot in males ［J］. High Alt Med Biol ， 2015 ， 16 （ 1 ）： 52 - 60 .

顾兵，金建波，李华南，等 . 神经组织染色方法的研究概况［J］. 中国药理学通报， 2011 ， 27 （ 10 ）： 1472 - 1475 .

COULTER D A ， EID T . Astrocytic regulation of glutamate homeostasis in epilepsy ［J］. Glia ， 2012 ， 60 （ 8 ）： 1215 - 1226 .

GREWER C ， GAMEIRO A ， ZHANG Z ， et al . Glutamate forward and reverse transport：from molecular mechanism to transporter-mediated release after ischemia ［J］. IUBMB Life ， 2008 ， 60 （ 9 ）： 609 - 619 .

BALAZS L . Involvement of extrasynaptic glutamate in physiological and pathophysiological changes of neuronal excitability ［J］. Cell Mol Life Sci ， 2018 ， 75 （ 16 ）： 2917 - 2949 .

BANASR M ， CHOWDHURY G M ， TERWILLIGER R ， et al . Glial pathology in an animal model of depression：reversal of stress-induced cellular，metabolic and behavioral deficits by the glutamate-modulating drug riluzole ［J］. Mol Psychiatry ， 2010 ， 15 （ 5 ）： 501 - 511 .

MIGUEL-HIDALGO J J ， WALTZER R ， WHITTOM A A ， et al . Glial and glutamatergic markers in depression，alcoholism，and their comorbidity ［J］. J Affect Disord ， 2010 ， 127 （ 1/3 ）： 230 - 240 .

BERNARD R ， KERMAN I A ， THOMPSON R C ， et al . Altered expression of glutamate signaling，growth factor，and glia genes in the locus coeruleus of patients with major depression ［J］. Mol Psychiatry ， 2011 ， 16 （ 6 ）： 634 - 646 .

MIYAGISHI H ， TSUJI M ， SAITO A ， et al . Inhibitory effect of yokukansan on the decrease in the hippocampal excitatory amino acid transporter EAAT2 in stress-maladaptive mice ［J］. J Tradit Complement Med ， 2017 ， 7 （ 4 ）： 371 - 374 .

ZINK M ， VOLLMAYR B ， GEBICKE-HAERTER P J ， et al . Reduced expression of glutamate transporters vGluT1，EAAT2 and EAAT4 in learned helpless rats，an animal model of depression ［J］. Neuropharmacology ， 2010 ， 58 （ 2 ）： 465 - 473 .

徐雅岚，赵洪庆，杜青，等 . 左归降糖解郁方对糖尿病并发抑郁症大鼠海马区星形胶质细胞谷氨酸转运体的影响［J］. 北京中医药大学学报， 2016 ， 39 （ 6 ）： 470 - 475 .

葛渴敏，王薇，薛文达，等 . 越鞠丸合甘麦大枣汤加减对谷氨酸诱导的HT22细胞损伤模型的神经保护作用［J］. 中国实验方剂学杂志， 2019 ， 25 （ 12 ）： 22 - 27 .

许鹏，章程鹏 . 基于抑郁症发病机制的中药防治规律［J］. 中国实验方剂学杂志， 2020 ， 26 （ 7 ）： 232 - 238 .

郭延红，夏忠玉，陈江，等 . 人参皂苷Rg 1 对慢性应激抑郁模型大鼠谷氨酸及其受体表达的影响［J］. 中国医院药学杂志， 2019 ， 39 （ 2 ）： 137 - 141 .

YOSHIZUMI M ， EISENACH J C ， HAYASHIDA K . Riluzole and gabapentinoids activate glutamate transporters to facilitate glutamate-induced glutamate release from cultured astrocytes ［J］. Eur J Pharmacol ， 2012 ， 677 （ 1/3 ）： 87 - 92 .

CARBONE M ， DUTY S ， RATTRAY M . Riluzole elevates GLT-1 activity and levels in striatal astrocytes ［J］. Neurochem Int ， 2012 ， 60 （ 1 ）： 31 - 38 .

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