从BDNF/TrkB信号通路探讨乌头汤对神经病理性疼痛模型小鼠脑神经元的保护作用

师钰琪; 吴红艳; 朱春燕; 林娜

doi:10.13422/j.cnki.syfjx.20200639

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从BDNF/TrkB信号通路探讨乌头汤对神经病理性疼痛模型小鼠脑神经元的保护作用

经典名方 | 更新时间：2021-02-09

- 从BDNF/TrkB信号通路探讨乌头汤对神经病理性疼痛模型小鼠脑神经元的保护作用
- Neuroprotective Effect by Wutoutang in Spinal Nerve Ligation Mice via BDNF/TrkB Signaling Pathway
- 中国实验方剂学杂志 2020年26卷第7期页码：23-30
- 作者机构：
  
  1.广州中医药大学，广州　 510006
  2.中国中医科学院　中药研究所，北京　 100700
- 作者简介：
  
  [第一作者]　师钰琪，在读硕士，从事中药药理学研究，E-mail：qq790863493@163.com
  *朱春燕，博士，从事中药药理学研究，E-mail：xijiangyue3013@163.com；
  *林娜，博士，研究员，从事中药药理学研究，E-mail：linna888@163.com
- 基金信息：
  
  国家“重大新药创制”科技重大专项(2019ZX09731002);中央级公益性科研院所基本科研业务费课题(L2017046);国家自然科学基金项目(81630107;81603322)
- DOI：10.13422/j.cnki.syfjx.20200639
  中图分类号： R2-0; R289; R285.5
- 收稿日期：2019-09-02，
  
  网络出版日期：2019-12-09，
  
  纸质出版日期：2020-04-05
- 稿件说明：
移动端阅览
师钰琪, 吴红艳, 朱春燕, 等. 从BDNF/TrkB信号通路探讨乌头汤对神经病理性疼痛模型小鼠脑神经元的保护作用[J]. 中国实验方剂学杂志, 2020,26(7):23-30.

Yu-qi SHI, Hong-yan WU, Chun-yan ZHU, et al. Neuroprotective Effect by Wutoutang in Spinal Nerve Ligation Mice via BDNF/TrkB Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(7): 23-30.
师钰琪, 吴红艳, 朱春燕, 等. 从BDNF/TrkB信号通路探讨乌头汤对神经病理性疼痛模型小鼠脑神经元的保护作用[J]. 中国实验方剂学杂志, 2020,26(7):23-30. DOI： 10.13422/j.cnki.syfjx.20200639.

Yu-qi SHI, Hong-yan WU, Chun-yan ZHU, et al. Neuroprotective Effect by Wutoutang in Spinal Nerve Ligation Mice via BDNF/TrkB Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(7): 23-30. DOI： 10.13422/j.cnki.syfjx.20200639.

摘要

目的：

从脑源性神经营养因子(BDNF)/酪氨酸激酶受体B(TrkB)信号通路研究乌头汤对神经病理性疼痛模型小鼠的神经元保护作用。

方法：

40只雄性ICR小鼠随机分为假手术组(Sham)，SNL模型组，乌头汤组(126 mg·kg

-1

)，ANA-12＋乌头汤组。建立神经病理性疼痛L5脊神经结扎(SNL)小鼠模型，造模成功后，灌胃给药10 d，脑立体定位注射BDNF/TrkB受体拮抗剂ANA-12(50 nmol·L

-1

)7 d，免疫组化法检测小鼠海马BDNF，蛋白激酶B(Akt)，环磷酸腺苷(cAMP)反应元件结合蛋白(CREB)蛋白的表达。免疫荧光法观察谷氨酸(Glu)和

-氨基丁酸(GABA)神经元变化。小鼠海马神经元细胞分离于精准孕18 d的胎鼠脑组织，分为正常组、甘氨酸处理组，ANA-12组(0.5 mmol·L

-1

)，ANA-12＋甘氨酸组，ANA-12＋乌头汤组，ANA-12＋乌头汤＋甘氨酸组，细胞免疫荧光法观察原代神经元形态学变化；海马神经元细胞分为正常组、甘氨酸处理组(200 mmol·L

-1

)，肿瘤坏死因子(TNF)-

(5 mg·L

-1

)＋甘氨酸组，TNF-

＋乌头汤(5 mg·L

-1

)＋甘氨酸组，TNF-

＋乌头汤＋甘氨酸＋BDNF-siRNA组，TNF-

＋乌头汤＋甘氨酸＋Akt-siRNA组，TNF-

＋乌头汤＋甘氨酸＋CREB-siRNA组，细胞免疫荧光法检测神经元初、次级树突棘内外的突触后密度蛋白95(PSD95)的表达量。

结果：

与正常组比较，模型组BDNF，Akt和CREB阳性细胞数显著下降(

0.01)，Glu-GABA能神经元失衡(

0.01)；与模型组比较，乌头汤组BDNF，Akt和CREB阳性细胞数显著上升(

0.01)，Glu-GABA能神经元恢复平衡(

0.01)；与乌头汤组比较，ANA-12＋乌头汤组BDNF和CREB阳性细胞数又显著下降(

0.05)而Akt组并无显著变化，Glu-GABA能神经元失衡(

0.01)。与正常组比较，甘氨酸处理组神经元初、次级树突棘数量显著上升(

0.01)，且突触后密度蛋白95(PSD95)表达量显著上升(

0.01)，ANA-12组，ANA-12＋甘氨酸组，ANA-12＋乌头汤组，ANA-12＋乌头汤＋甘氨酸组神经元初、次级树突棘数量均无明显变化；与甘氨酸处理组比较，TNF-

＋甘氨酸组PSD95表达量显著下降(

0.01)；与TNF-

＋甘氨酸组比较，TNF-

＋乌头汤＋甘氨酸组PSD95表达量明显上升(

0.01)；与TNF-

＋乌头汤＋甘氨酸组比较，TNF-

＋乌头汤＋甘氨酸＋BDNF-siRNA组，TNF-

＋乌头汤＋甘氨酸＋Akt-siRNA组，TNF-

＋乌头汤＋甘氨酸＋CREB-siRNA组PSD95表达量均显著下降(

0.01)。

结论：

乌头汤对SNL小鼠海马谷氨酸能神经元兴奋性及可塑性损伤具有修复作用，这一作用与其对BDNF/TrkB通路的调控有关。

Abstract

Objective:

To investigate the neuroprotective effect and mechanism by Wutoutang (WTT) in the spinal nerve ligation (SNL) mice by neurotrophic factor (BDNF)/tyrosine kinase receptor B (TrkB) signaling BDNF/tyrosine kinase receptor B (TrkB).

Method:

The 40 mice were randomly divided into Sham group

SNL group

WTT group(126 mg·kg

-1

)

ANA-12+ WTT group.The L5 spinal nerve ligation model mice were established in mice

After that

WTT was administrated from the first day to the 10

day

then the consecutive 7-day hippocampal injection of ANA-12(0.05 nmol·L

-1

)were lasted for 7 days.The levels of brain-derived BDNF

cAMP-response element binding protein(CREB)

and protein kinase B(Akt)and the change of hippocampal glutamatergic and GABAergic neurons in mice were detected by tissue immunofluorescence.E14 pregnant ICR mice were sacrificed and the hippocampus were dissected which were divided into control group

glycine group

tumo necrosis factor(TNF)-

(5 mg·L

-1

)+ glycine group

TNF-

+ WTT(5 mg·L

-1

)+ glycine group

TNF-

+ WTT+ glycine+ BDNF-siRNA group

TNF-

+ WTT+ glycine+ Akt-siRNA group

TNF-

+ WTT+ glycine+ CREB-siRNA group

the primary and secondary dendrrictes

in which the arrowheads indicate the expression od postsynapti desity protein 95(PSD95) in the shafts and arrows were tested by cellular immunofluorescence.The neurons were divided into control group

glycine group

ANA-12 group(0.5 mmol·L

-1

)

ANA-12+ glycine group

ANA-12+ WTT group

ANA-12+ WTT+ glycine group

the morphology of hippocampal neurons were tested by cellular immunofluorescence.

Result:

Compared with Sham group

BDNF

Akt and CREB positive cell of SNL group decreased significantly(

0.01)

the hippocampal glutamatergic and GABAergic neurons out of balance(

0.01). Compared with SNL group

the BDNF

Akt and CREB positive cell of WTT group increased significantly(

0.01)

Glutamine-aminobutyric acid neurons regein balance(

0.01). Compared with WTT group

BDNF and CREB positive cell of ANA-12+ WTT group decreased significantly(

0.05)

Glutamine-aminobutyric acid neurons was disorderedd(

0.05). Comparaed with control group

the level of PSD95 of glycine group were increase significantly(

0.01). The number of dendritic spine density apically and basally of glycine group were increase significantly(

0.01)

but the primary and secondary dendrites of ANA-12 group

ANA-12+ glycine group

ANA-12+ WTT group

ANA-12+ WTT+ glycine group were not change.Comparaed with glycine group

the level of PSD95 of TNF-

+ glycine group were decreased significantly(

0.01). Comparaed with TNF-

+ glycine group

the level of PSD95 of TNF-

+ WTT+ glycine group were increase significantly(

0.01). Comparaed with TNF-

+ WTT+ glycine group

the level of PSD95 of TNF-

+ WTT+ glycine+ BDNF-siRNA group

TNF-

+ WTT+ glycine+ Akt-siRNA group

TNF-

+ WTT+ glycine+ CREB-siRNA group were decreased significantly(

0.01).

Conclusion:

In vivo

and

in vitro

studies have shown that the WTT mediated remission of the primary hippocampal glutamatergic neurons were dependent on the BDNF/TrkB pathway.

关键词

Keywords

references

T S JESEN , R BARON , M HAANPAA ， et al . A new definition of neuropathic pain [J]． Pain ， 2011 ， 152 ( 3 ): 2204 - 2205 .

F RADAT , A MARGOT-DUCLOT , N ATTAL ． Psychiatric co-morbidities in patients with chronic peripheral neuropathic pain：a multicenter cohort study [J]． Eur J Pain ， 2013 ， 17 ( 10 ): 1547 - 1557 .

K N ALSCHULER , D M EHDE , M P JENSEN . Co-occurring depression and pain in multiple sclerosis [J]. Phys Med Rehabil Clin N Am , 2013 ， 24 ( 4 ): 703 - 715 .

D BOUHASSIRA , M LANTERI-MINET , N ATTAL , et al . Prevalence of chronic pain with neuropathic characteristics in the general population [J]. Pain , 2008 ， 136 ( 3 ): 380 - 387 .

X Y LI , N WANG , Y J WANG ， et al . Long-term temporal imprecision of information coding in the anterior cingulate cortex of mice with peripheral inflammation or nerve injury [J]． J Neurosci ， 2014 ，( 32 ): 10675 - 10687 .

陈少珍．乌头汤治疗类风湿性关节炎的药效学及作用机理研究 [D]．广州：广州中医药大学， 2006 .

C Q LI , J M XU , D LIU ， et al . Brain derived neurotrophic factor(BDNF)contributes to the pain hypersensitivity following surgical incision in the rats [J]． Mol Pain ， 2008 ， 4 ( 27 ): 1 - 11 .

L T YI , J LI , B B LIU , et al . BDNF-ERK-CREB signalling mediates the role of miR-132 in the regulation of the effects of oleanolic acid in male mice [J]. J Psychiatry Neurosci , 2014 ， 39 ( 5 ): 348 - 359 .

吴红艳，师钰琪，朱春燕，等．乌头汤调控海马BDNF/TrkB通路以缓解神经病理性疼痛的痛共情绪症状 [J]．中国实验方剂学杂志， 2019 ， 26 ( 1 ): 24 - 30 .

C ZHU , Q XU , Z MAO , et al . The Chinese medicine Wu-Tou decoction relieves neuropathic pain by inhibiting hippocampal microglia activation [J]. Sci Rep , 2018 ， 8 ( 1 ): 12292 .

C ZHU , Q XU , C WANG , et al . Evidence that CA3 is underling the comorbidity between pain and depression and the co-curation by Wu-tou decoction in neuropathic pain [J]. Sci Rep , 2017 ， 7 ( 1 ): 11935 .

H X GE , S GUAN , Y L SHEN ， et al . Dihydromyricetin affects BDNF levels in the nervous system in rats with comorbid diabetic neuropathic pain and depression [J]． Sci Rep ， 2019 ， 9 ( 1 ): 14619 .

S KIM , J CHUNG ． An experimental model for peripheral neuropathy produced by segmental spinal nerve ligation in the rat [J]． Pain ， 1992 ， 50 ( 3 ): 355 - 363 .

S R CHAPLAN , F W BACH , J W POGREL ， et al . Quantitative assessment of tactile allodynia in the rat paw [J]． J Neurosci Methods ， 1994 ， 53 ( 1 ): 55 - 63 .

A R SANTOS , D COMPRIDO , C B DUARTE . Regulation of local translation at the synapse by BDNF [J]. Prog Neurobiol , 2010 , 92 ( 4 ): 505 - 516 .

H GE , S GUAN , Y SHEN , et al . Dihydromyricetin affects BDNF levels in the nervous system in rats with comorbid diabetic neuropathic pain and depression [J]. Sci Rep , 2019 ， 9 ( 1 ): 14619 .

F CONTI , R J WEINBERG . Shaping excitation at glutamatergic synapses [J]. Trends Neurosci , 1999 , 22 ( 10 ): 451 .

E CHERUBINI , F CONTI . Generating diversity at GABAergic synapses [J]. Trends Neurosci , 2001 , 24 ( 3 ): 155 .

D WANG , B LI , Y WU , et al . The effects of maternal atrazine exposure and swimming training on spatial learning memory and hippocampal morphology in offspring male rats via psd95/nr2b signaling pathway [J]. Cell Mol Neurobiol , 2019 ， 39 ( 7 ): 1003 - 1015 .

D WANG , B LI , Y WU , et al . The effects of maternal atrazine exposure and swimming training on spatial learning memory and hippocampal morphology in offspring male rats via PSD95/NR2B signaling pathway [J]. Cell Mol Neurobiol , 2019 ， 39 ( 7 ): 1003 - 1015 .

C L BARRETO , J R CARRILLO , G ROLDAN-ROLDAN , et al . Cyclic changes and actions of progesterone and allopregnanolone on cognition and hippocampal basal (stratum oriens) dendritic spines of female rats [J]. Behav Brain Res , 2019 : 112355 .

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