地黄饮子对脑缺血再灌注损伤大鼠保护作用及其机制

王俊杰; 楼琦; 汤娟娟; 白亚平; 李倩; 张培璐; 杜月光

doi:10.13422/j.cnki.syfjx.20190406

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地黄饮子对脑缺血再灌注损伤大鼠保护作用及其机制

经典名方 | 更新时间：2021-02-09

- 地黄饮子对脑缺血再灌注损伤大鼠保护作用及其机制
- Protective Effect of Dihuang Yinzi on Cerebral Ischemia-reperfusion Injury in Rats and Its Mechanism
- 中国实验方剂学杂志 2019年25卷第4期页码：42-48
- 作者机构：
  
  1.浙江中医药大学，杭州　 310053；
  2.浙江省医学科学院，杭州　 310013
- 作者简介：
  
  王俊杰，硕士，教授，从事中医药防治缺血性脑血管病及其机制研究，Tel：0571-86633390，E-mail：wjjie2000@163.com
  *杜月光，博士，教授，从事中医药防治缺血性脑血管病及其机制研究，Tel：0571-86633140，E-mail：1323944178@ qq.com
- 基金信息：
  
  浙江省自然科学基金项目(LY14H270005)
- DOI：10.13422/j.cnki.syfjx.20190406
  中图分类号： R2-0;R22;R285.5
- 纸质出版日期：2019-02-20，
  
  网络出版日期：2018-11-06，
  
  收稿日期：2018-08-13，
- 稿件说明：
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王俊杰, 楼琦, 汤娟娟, 等. 地黄饮子对脑缺血再灌注损伤大鼠保护作用及其机制[J]. 中国实验方剂学杂志, 2019,25(4):42-48.

Jun-jie WANG, Qi LOU, Juan-juan TANG, et al. Protective Effect of Dihuang Yinzi on Cerebral Ischemia-reperfusion Injury in Rats and Its Mechanism[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2019,25(4):42-48.
王俊杰, 楼琦, 汤娟娟, 等. 地黄饮子对脑缺血再灌注损伤大鼠保护作用及其机制[J]. 中国实验方剂学杂志, 2019,25(4):42-48. DOI： 10.13422/j.cnki.syfjx.20190406.

Jun-jie WANG, Qi LOU, Juan-juan TANG, et al. Protective Effect of Dihuang Yinzi on Cerebral Ischemia-reperfusion Injury in Rats and Its Mechanism[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2019,25(4):42-48. DOI： 10.13422/j.cnki.syfjx.20190406.

摘要

目的：

探讨地黄饮子对脑缺血再灌注损伤大鼠的保护作用及其初步机制。

方法：

建立大脑中动脉栓塞(MCAO)大鼠模型，随机分为6组，分别为假手术组，模型组，尼莫地平组(0.01 g·kg

－1

)，地黄饮子高、中、低剂量组(38.80，19.40，9.70 g·kg

－1

)，每组20只，连续给药28 d；术后第7，14，28天通过改良神经功能缺损评分(mNSS)，第28天行2，3，5-氯化三苯基四氮唑(TTC)染色，苏木素-伊红(HE)染色观察地黄饮子的保护作用，应用免疫荧光技术观察侧脑室室管膜下区(SVZ)的5-溴脱氧尿嘧啶核苷(BrdU)阳性细胞数(第28天)以分析神经干细胞(NSCs)增殖情况，采用实时荧光定量聚合酶链式反应(Real-time PCR)观察Notch1，Jagged1，Hes1 mRNA的表达情况。

结果：

术后第7，14，28天地黄饮子高、中剂量组，尼莫地平组的mNSS明显低于同时间段的模型组(

0.05，

0.01)；术后第28天地黄饮子高、中剂量组和尼莫地平组的脑梗死体积占脑组织体积的百分比均低于模型组(

0.01)；HE染色显示地黄饮子高、中剂量组，尼莫地平组大鼠脑组织坏死和软化灶不明显，并可见神经元细胞和神经胶质细胞增生；术后第28天，上述3组SVZ的BrdU阳性细胞数显著高于模型组(

0.05，

0.01)，地黄饮子高剂量组BrdU阳性细胞数高于尼莫地平组(

0.05)；术后28 d地黄饮子高、中剂量组，尼莫地平组SVZ的Notch1，Jagged1，Hes1 mRNA表达水平明显高于模型组(

0.05，

0.01)；尼莫地平组Hes1 mRNA水平高于地黄饮子高、中、低剂量组(

0.05，

0.01)。

结论：

地黄饮子能改善MCAO大鼠模型的神经功能缺损，减小脑梗死体积，减轻脑组织病理改变，从而对脑缺血再灌注损伤大鼠起到保护作用，可能与激活Notch信号通路，上调Notch1，Jagged1，Hes1 mRNA的表达，从而促进NSCs增殖有关。

Abstract

Objective:

To explore the protective effect and the preliminary mechanism of Dihuang Yinzi on cerebral ischemia-reperfusion injury in rats.

Method:

The middle cerebral artery occlusion (MCAO) model was established. Totally 90 SD rats were randomly divided into 6 groups: sham operation group

model group

nimodipine group (0.01 g·kg

-1

) and high

medium

low-dose Dihuang Yinzi groups (38.80

19.40

9.70 g·kg

-1

)

with 20 rats in each group.The modified neurological severity score (mNSS) was assayed at the 7

days after operation

and the volume of cerebral infarction

pathological changes of brain tissue

the BrdU positive cells and mRNA levels of Notch1

Jagged1 and Hes1 in subventricular zone(SVZ)were observed respectively by triphenyl tetrazolium chloride(TTC) stain

htorylin eastin(HE) stain

immunofluorescence technique and reverse transcriphase polymerase chain reaction(Real-time PCR) methods at the 28

day after the operation.

Result:

The mNSS on the 7

days of high

medium-dose Dihuang Yinzi groups and nimodipine group were significantly lower than that of model group(

0.05

0.01). On the 28

day

the percentage of cerebral infarction volume in brain tissue volume of high

medium-dose Dihuang Yinzi groups and nimodipine group were smaller than that of model group(

0.01). HE staining showed that the necrosis and softening lesions of brain tissue were not obvious in rats of high

medium-dose Dihuang Yinzi groups and nimodipine group

with neuronal cell and neuroglial cell proliferation. On the 28

day

the BrdU positive cells in SVZ of the above 3 groups were significantly higher than model group(

0.05

0.01)

and the high-dose Dihuang Yinzi group was significantly higher than nimodipine group(

0.05). On the 28

day

the mRNA levels of Notch1

Jagged1 and Hes1 of high

medium-dose Dihuang Yinzi groups and nimodipine group were significantly higher than those of model group(

0.05

0.01)

the mRNA level of Hes1 of nimodipine group was higher than that of high

medium

low-dose Dihuang Yinzi groups(

0.05

0.01).

Conclusion:

Dihuang Yinzi can improve the nerve function defect of MCAO rat model

and reduce the volume of cerebral infarction and the pathological changes of brain tissue

thus playing a protective role in cerebral ischemia-reperfusion injury rats. Its mechanism may be related to the activation of the Notch signaling pathway

and the up-regulation of expressions of Notch1

Jagged1 and Hes1 mRNA

thus promoting the proliferation of NSCs.

关键词

地黄饮子脑缺血再灌注损伤实验研究神经干细胞

Keywords

Dihuang Yinzicerebral ischemia-reperfusion injuryexperimental study;neural stem cells

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