Effect of Modified Chaihu Shugantang Regulat miRNA-204 on Hippocampus Autophagy in Epileptic Mice

Shun-gui WANG; Qian YU; Hua-xia LI; Huan LI; Ling LU; Xian-qiu LIAO; Qiong WU; Hua-qiong LI; Ai-ling CHEN; Li-mei DIAO; Qian-chao HE; Lun CAI; Shuang LIANG

doi:10.13422/j.cnki.syfjx.20192137

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Effect of Modified Chaihu Shugantang Regulat miRNA-204 on Hippocampus Autophagy in Epileptic Mice

Pharmacology | 更新时间：2021-02-09

- Effect of Modified Chaihu Shugantang Regulat miRNA-204 on Hippocampus Autophagy in Epileptic Mice
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 22, Pages: 1-7(2019)
- 作者机构：
  
  1.广西中医药大学　研究生学院，南宁　 530023；
  2.广西中医药大学　第一附属医院，南宁　 530001
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20192137
  CLC： R2-0; R285; R289; R742.1
- Published：20 November 2019，
  
  Published Online：18 July 2019，
  
  Received：02 April 2019，
- 稿件说明：
扫描看全文
Shun-gui WANG, Qian YU, Hua-xia LI, et al. Effect of Modified Chaihu Shugantang Regulat miRNA-204 on Hippocampus Autophagy in Epileptic Mice. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(22):1-7(2019)
DOI：

Shun-gui WANG, Qian YU, Hua-xia LI, et al. Effect of Modified Chaihu Shugantang Regulat miRNA-204 on Hippocampus Autophagy in Epileptic Mice. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(22):1-7(2019) DOI： 10.13422/j.cnki.syfjx.20192137.

摘要

目的：

观察加味柴胡疏肝汤对癫痫小鼠海马微小非编码RNA 204(miRNA-204)的表达影响，探讨其发挥神经保护作用的机制。

方法：

将60只小鼠随机分为6组：正常组，模型组(盐酸匹罗卡品180 mg·kg

－1

)，加味柴胡疏肝汤组(7 g·kg

－1

·d

－1

)，加味柴胡疏肝汤＋miRNA-204模拟物组(7 g·kg

－1

·d

－1

＋2 μL)，加味柴胡疏肝汤＋miRNA-204抑制物组(7 g·kg

－1

·d

－1

＋2 μL)，卡马西平组(30 mg·kg

－1

·d

－1

)，各灌胃2周；采用匹罗卡品使小鼠致痫，6组分别灌胃给药、抑制及过表达miRNA-204后，处死小鼠，取其海马组织，观察每组的指标情况，采用实时荧光定量聚合酶链式反应(Real-time PCR)检测小鼠海马miRNA-204的表达；蛋白免疫印迹法(Western blot)检测自噬相关蛋白微管相关蛋白轻链3(LC3)，自噬相关标志性蛋白7(ATG7)的表达情况，苏木素-伊红(HE)染色观察每组小鼠海马病理学变化；透射电镜观察各组小鼠海马组织自噬情况。

结果：

与正常组比较，模型组小鼠miRNA-204表达显著降低(

0.01)，海马C1区病理改变最明显，ATG7，LC3Ⅱ/LC3Ⅰ表达显著升高(

0.01)，自噬小体数量最多；与模型组比较，加味柴胡疏肝组小鼠海马组织miRNA-204表达明显升高(

0.05)，海马C1区病理改变减轻，ATG7，LC3Ⅱ/LC3Ⅰ表达明显降低(

0.05)，自噬小体数量减少；加味柴胡疏肝汤＋miRNA-204模拟物组小鼠海马组织miRNA-204表达显著升高(

0.01)，海马C1区病理改变最轻，ATG7，LC3Ⅱ/LC3Ⅰ表达显著降低(

0.01)，自噬小体数量最少；与加味柴胡疏肝汤比较，加味柴胡疏肝汤＋miRNA-204抑制物组上述指标变化趋势相同，变化幅度明显减少(

0.05)。

结论：

加味柴胡疏肝汤可以改善癫痫小鼠海马组织病理改变，其作用机制可能是通过升高癫痫小鼠海马miRNA-204的表达，抑制神经元过度自噬，减少细胞凋亡。

Abstract

Objective:

To observe the effect of modified Chaihu Shugantang on the expression of miRNA-204 in hippocampus of epileptic mice

and to explore its mechanism of neuroprotection.

Method:

The sixty mice were randomly divided into 6 groups: normal group

model group (pilocarpine 180 mg·kg

-1

)

and modified Chaihu Shugantang group (7 g·kg

-1

·d

-1

)

modified Chaihu Shugantang+ miRNA-204 mimic group (7 g·kg

-1

·d

-1

+ 2 μL)

modified Chaihu Shugantang+ miRNA-204 inhibitor group (7 g·kg

-1

·d

-1

+ 2 μL)

carbamazepine group (30 mg·kg

-1

·d

-1

)

each was given intragastric administration for 2 weeks

using pilocarpine to cause epilepsy in mice

respectively

add flavor to Bupleurum after intragastric administration

inhibition and overexpression of miRNA-204

the mice were sacrificed and their hippocampus tissues were harvested.The indicators of each group were observed

Real-time quantitative PCR detecting system (Real-time PCR) was used to detect mouse hippocampal miRNA-204 expression

Western blot analysis of autophagy-related protein microtubule-associated protein light chain 3 (LC3)

autophagy-associated marker protein 7 (ATG7) expression

hematoxylin pathological condition of hippocampus in each group was observed by hematoxylin-eosin(HE)staining.The autophagy of hippocampus in each group was observed by transmission electron microscopy.

Result:

Compared with normal group

the expression of miRNA-204 was significantly decreased in model group (

0.01)

the pathological changes in the hippocampal C1 area were the most obvious

the expression of ATG7

LC3Ⅱ/LC3Ⅰ was increased (

0.01)

and the autophagy was small. Compared with model group

the expression of miRNA-204 in the hippocampus of the modified Chaihu Shugantang group was increased (

0.05)

the pathological changes in the hippocampal C1 area were alleviated

the expression of ATG7

LC3Ⅱ/LC3Ⅰ was decreased (

0.05)

and the autophagy was small. The number of body decreased

the expression of miRNA-204 in hippocampus of modified Chaihu Shugantang+ miRNA-204 mimic group was significantly increased (

0.01)

the pathological changes in hippocampal C1 area were the lightest

and the expression of ATG7

LC3II/LC3I was decreased (

0.01)

the number of autophagosomes was the least.Compared with modified Chaihu Shugantang group

the above-mentioned indicators of modified Chaihu Shugantang+ miRNA-204 inhibitor group had the same change trend and the change range decreased (

0.05).

Conclusion:

Modified Chaihu Shugantang can improve the pathological changes of hippocampus in mice with epilepsy and play a neuroprotective role. The mechanism may be to increase the expression of miRNA-204 in hippocampus of mice with epilepsy

inhibit excessive autophagy of neurons and reduce apoptosis.

关键词

柴胡疏肝汤癫痫miRNA-204自噬

Keywords

Chaihu ShugantangepilepsymiRNA-204autophagy

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