基于Keap1/Nrf2通路探讨补肾健脾开心方对衰老大鼠学习认知能力及脑皮质氧化应激的影响

李莹; 王莹; 刘琳; 王晓瑞; 孔明望

doi:10.3969/j.issn.1006-2157.2023.09.010

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基于Keap1/Nrf2通路探讨补肾健脾开心方对衰老大鼠学习认知能力及脑皮质氧化应激的影响

实验研究 | 更新时间：2023-10-09

- 基于Keap1/Nrf2通路探讨补肾健脾开心方对衰老大鼠学习认知能力及脑皮质氧化应激的影响
- Effect of Bushen Jianpi Kaixin Formula on learning and cognitive ability and oxidative stress in the cerebral cortex of aging rats through the Keap1/Nrf2 pathway
- 北京中医药大学学报 2023年46卷第9期页码：1250-1257
- 作者机构：
  
  湖北中医药大学基础医学院　武汉　430065
- 作者简介：
  
  李莹，女，在读博士生
  # 孔明望，男，博士，教授，博士生导师，主要研究方向：中医衰老理论及老年病防治，E-mail：13349959661@163.com
- 基金信息：
  
  国家重点研发计划“中医药现代化研究”重点专项(2019YFC1708502)
- DOI：10.3969/j.issn.1006-2157.2023.09.010
  中图分类号： R285.5
- 收稿日期：2023-02-04，
  
  网络出版日期：2023-07-11，
  
  纸质出版日期：2023-09-30
- 稿件说明：
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李莹, 王莹, 刘琳, 等. 基于Keap1/Nrf2通路探讨补肾健脾开心方对衰老大鼠学习认知能力及脑皮质氧化应激的影响[J]. 北京中医药大学学报, 2023,46(9):1250-1257.

LI Ying, WANG Ying, LIU Lin, et al. Effect of Bushen Jianpi Kaixin Formula on learning and cognitive ability and oxidative stress in the cerebral cortex of aging rats through the Keap1/Nrf2 pathway[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(9): 1250-1257.
李莹, 王莹, 刘琳, 等. 基于Keap1/Nrf2通路探讨补肾健脾开心方对衰老大鼠学习认知能力及脑皮质氧化应激的影响[J]. 北京中医药大学学报, 2023,46(9):1250-1257. DOI： 10.3969/j.issn.1006-2157.2023.09.010.

LI Ying, WANG Ying, LIU Lin, et al. Effect of Bushen Jianpi Kaixin Formula on learning and cognitive ability and oxidative stress in the cerebral cortex of aging rats through the Keap1/Nrf2 pathway[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(9): 1250-1257. DOI： 10.3969/j.issn.1006-2157.2023.09.010.

摘要

目的

观察补肾健脾开心方对衰老大鼠学习记忆能力、认知功能及脑皮质氧化应激的影响，并探讨其作用机制。

方法

47只SD大鼠根据随机数字表法分为空白组(

＝10)和造模组(

＝37)。造模组大鼠腹腔注射D－半乳糖生理盐水溶液(300 mg/kg)，连续6周，建立衰老大鼠模型。造模成功后，任意取30只大鼠，随机数字表法分为模型组、补肾健脾开心方组和维生素E组，每组10只。补肾健脾开心方组大鼠给予补肾健脾开心方(9.99 g/kg)灌胃，维生素E组大鼠给予维生素E(13.5 mg/kg)灌胃，空白组、模型组大鼠给予等体积生理盐水灌胃，每日1次，连续4周。给药结束后，Morris水迷宫实验检测大鼠学习记忆能力，旷场实验检测大鼠认知功能，WST－1法检测超氧化物歧化酶(SOD)活力、TBA法检测丙二醛(MDA)含量、比色法检测抑制羟自由基能力，免疫组织化学法检测脑皮质半胱天冬酶－1(Caspase－1)阳性表达，实时荧光PCR法检测脑皮质Kelch样环氧氯丙烷相关蛋白1(Keap1)、核红细胞2相关因子2(Nrf2)、醌氧化还原酶1(NQO1)、血红素加氧酶－1(HO－1)mRNA表达。

结果

与空白组比较，模型组大鼠逃避潜伏期从第3天开始延长(

0.05，

0.01)、跨越平台次数减少(

0.01)，运动距离和中心区域运动时间比例减少(

0.01)，SOD活力和抑制羟自由基能力减弱(

0.01)，MDA表达增多(

0.01)，Caspase－1阳性表达增多(

0.01)，Keap1 mRNA表达增多(

0.01)，Nrf2、NQO1、HO－1 mRNA表达减少(

0.05，

0.01)。与模型组比较，补肾健脾开心方组大鼠第4、5天逃避潜伏期缩短(

0.05，

0.01)、跨越平台次数增多(

0.01)，运动距离和中心区域运动时间比例增多(

0.01)，SOD活力和抑制羟自由基能力增强(

0.01)，MDA表达减少(

0.01)，Caspase－1阳性表达减少(

0.01)，Keap1 mRNA表达减少(

0.01)，Nrf2、HO－1 mRNA表达增多(

0.05，

0.01)。

结论

补肾健脾开心方可以改善衰老大鼠认知障碍、延缓衰老，其作用机制可能与调控氧化应激有关。

Abstract

Objective

To observe the effects of the

Bushen Jianpi Kaixin

Formula(BJKF) on the learning and memory ability

cognitive function

and oxidative stress in the cerebral cortex of aging rats

and to explore its mechanism.

Methods

According to the random number table method

47 Sprague-Dawley rats were randomly divided into the blank group(

=10) and the modeling group(

=37). The rats in the modeling group were injected with D-galactose normal saline solution(300 mg/kg) for 6 weeks to establish the aging rat model. After successful modeling

according to the random number table method

30 aging rats were randomly divided into the model group

BJKF group and vitamin E group

with 10 rats each group. The rats in the BJKF group were treated with BJKF(9.99 g/kg) by gavage

the rats in the vitamin E group were treated with vitamin E(13.5 mg/kg) by gavage

and the rats in the blank group and the model group were treated with the same volume of normal saline by gavage. All the interventions were administered once daily for 4 weeks. After the intervention

the learning and memory abilities of rats were detected by the Morris water maze test. The cognitive function of rats was detected by the open-field test. The activity of superoxide dismutase(SOD) was detected by the WST-1 method. The content of malondialdehyde(MDA) was determined by TBA method. The hydroxyl radical inhibition ability was detected by colorimetry. The positive expression of Caspase-1 in the cerebral cortex was detected by immunohistochemistry. And the mRNA expressions of Keap1

Nrf2

NQO1 and HO-1 were detected by real-time PCR.

Results

Compared with the blank group

the evasion latency was prolonged from the third day(

0.05

0.01)

the number of plateau crossing was decreased(

0.01)

the movement distance and the time ratio of central area movement were decreased (

0.01)

the SOD activity and the hydroxyl radical inhibition ability were decreased(

0.01)

the expression of MDA was increased (

0.01)

the positive expression of Caspase-1 was increased(

0.01)

the mRNA expression of Keap1 was increased(

0.01)

and the mRNA expressions of Nrf2

NQO1

and HO-1 were decreased(

0.05

0.01) in the model group. Compared with the model group

the evasion latency was shortened on the fourth and fifth day(

0.05

0.01)

the number of plateau crossing was increased(

0.01)

the movement distance and the time ratio of central area movement were increased (

0.01)

the SOD activity and the hydroxyl radical inhibition ability were increased(

0.01)

the expression of MDA was decreased(

0.01)

the positive expression of Caspase-1 was decreased(

0.01)

the mRNA expression of Keap1 was decreased(

0.01)

while the mRNA expressions of Nrf2 and HO-1 were increased(

0.05

0.01) in the BJKF group.

Conclusion

BJKF can improve cognitive impairment and delay aging in aging rats whose mechanism is thought to be related to regulating oxidative stress.

关键词

Keywords

references

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