天王补心丹加减改善PCPA失眠大鼠Trx系统氧化损伤的机制探讨

谢光璟; 黄攀攀; 王平

doi:10.13422/j.cnki.syfjx.20190637

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天王补心丹加减改善PCPA失眠大鼠Trx系统氧化损伤的机制探讨

经典名方 | 更新时间：2021-02-09

- 天王补心丹加减改善PCPA失眠大鼠Trx系统氧化损伤的机制探讨
- Mechanism of Modified Tianwang Buxindan on Trx System Oxidative Damage in PCPA Insomnia Model Rats
- 中国实验方剂学杂志 2019年25卷第6期页码：32-38
- 作者机构：
  
  湖北中医药大学　基础医学院，武汉　 430065
- 作者简介：
  
  谢光璟，在读博士，从事中医药防治老年病研究，E-mail：397525306@qq.com
  王平，教授，主任医师，博士生导师，从事中医衰老理论及老年病证治疗规律研究，E-mail：pwang54@163.com
- 基金信息：
  
  国家自然科学基金面上项目(81573865;81874414);湖北省首届医学领军人才培养工程项目(5406-0063)
- DOI：10.13422/j.cnki.syfjx.20190637
  中图分类号： R289;R289.5;R256.23
- 收稿日期：2018-10-16，
  
  网络出版日期：2018-06-12，
  
  纸质出版日期：2019-03-20
- 稿件说明：
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谢光璟, 黄攀攀, 王平. 天王补心丹加减改善PCPA失眠大鼠Trx系统氧化损伤的机制探讨[J]. 中国实验方剂学杂志, 2019,25(6):32-38.

Guang-jing XIE, Pan-pan HUANG, Ping WANG. Mechanism of Modified Tianwang Buxindan on Trx System Oxidative Damage in PCPA Insomnia Model Rats[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(6): 32-38.
谢光璟, 黄攀攀, 王平. 天王补心丹加减改善PCPA失眠大鼠Trx系统氧化损伤的机制探讨[J]. 中国实验方剂学杂志, 2019,25(6):32-38. DOI： 10.13422/j.cnki.syfjx.20190637.

Guang-jing XIE, Pan-pan HUANG, Ping WANG. Mechanism of Modified Tianwang Buxindan on Trx System Oxidative Damage in PCPA Insomnia Model Rats[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(6): 32-38. DOI： 10.13422/j.cnki.syfjx.20190637.

摘要

目的：

探讨天王补心丹加减对对氯苯丙氨酸(PCPA)所致失眠大鼠硫氧还蛋白(Trx)系统抗氧化损伤的作用机制。

方法：

60只雄性SD大鼠随机分为空白组、造模组。造模组以腹腔注射PCPA造模，注射剂量为150 mg·kg

－1

，间断注射7 d。造模成功后，依次分为模型组（等容生理盐水），天王补心丹加减低、中、高剂量组（8.8，17.6，35.2 g·kg

－1

·d

－1

）和艾司唑仑组（0.1 mg·kg

－1

·d

－1

），每组10只。自发活动视频分析系统监测大鼠昼夜活动节律，透射电镜观察大鼠下丘脑视交叉上核（SCN）形态及细胞器完整性，酶联免疫吸附测定法（ELISA）检测大鼠血清超氧化物歧化酶（SOD），丙二醛（MDA）和谷胱甘肽过氧化物酶（GSH-Px）的水平，免疫荧光（IF）和蛋白免疫印迹法（Western blot）分别检测SCN细胞硫氧环蛋白2（Trx2），硫氧环蛋白还原酶2（TrxR2）的表达。

结果：

与空白组比较，模型组大鼠活动时间显著增加(

0.01)，昼夜节律紊乱，线粒体嵴断裂，数量减少，排列紊乱，伴空泡化和髓鞘样变，SOD，GSH-Px活性下降且MDA含量显著升高(

0.01)，Trx2的蛋白表达量显著降低(

0.01)，TrxR2蛋白表达量显著增加(

0.01)；与模型组比较，天王补心丹中、高剂量组能够明显改善大鼠昼夜节律紊乱，减少活动时间(

0.01)，同时线粒体水肿程度减轻，部分嵴尚完整，SOD，GSH-Px活性升高，MDA水平明显降低(

0.05，

0.01)，Trx2蛋白表达明显升高，TrxR2蛋白表达明显下降(

0.05)。

结论：

天王补心丹加减对失眠模型大鼠的改善作用与调控Trx抗氧化系统中Trx2与TrxR2的蛋白表达有关。

Abstract

Objective:

To explore the mechanism of modified Tianwang Buxindan on oxidative damage of Trx system in parachlorophenylalanine (PCPA) insomnia model rats.

Method:

Sixty male SD rats were randomly divided into blank group and model group. Insomnia model was prepared through intraperitoneal injection with PCPA (150 mg·kg

-1

). The discontinuous injection lasted for 7 d. After successful modeling

the rats were divided into model group (the same volume of normal saline)

low

medium

high-dose Tianwang Buxindan groups (8.8

17.6

35.2 g·kg

-1

·d

-1

) and estazolam group (0.1mg·kg

-1

·d

-1

)

with 10 in each group. Autonomous activity video was used to detect circadian activity rhythm. Transmission electron microscopy (TEM) was used to observe supra chiasmatic nucleus (SCN) morphology and Organelle integrity. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expressions of superoxide dismutase (SOD)

malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in serum. Expressions of Trx2

TrxR2 in SCN cells were detected by immunofluorescence (IF) and Western blot.

Result:

Compared with blank group

the activity rhythm of model group was irregular

the activity time increased (

0.01)

the mitochondrial cristae were broken

the number was reduced

the arrangement was disorder

SOD and GSH-PX decreased

MDA increased (

0.01)

the expression of Trx2 decreased

while the expression of TrxR2 increased (

0.01). Compared with model group

medium and high-dose Tianwang Buxindan groups could alleviate the circadian rhythm disorder and reduce the activity time (

0.01)

the mitochondrial edema was relieved

part of the cristae were intact

the activities of SOD and GSH-PX were increased

the level of MDA was reduced (

0.05

0.01)

expression of Trx2 was significantly increased

and expression of TrxR2 was significantly reduced (

0.05

0.01).

Conclusion:

The effect of modified Tianwang Buxindan on insomnia model rats is related to the regulation of Trx2 and TrxR2 protein expressions in Trx system.

关键词

Keywords

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