Intervention Effect of Kidney-tonifying and Essence-filling Therapy on Hippocampal Autophagy in Kidney-deficiency Alzheimer Mice

Zhong-kang ZHU; Lin ZHANG; Chun LIU; Xu WANG; Hui-hui LIU; Dan-yu ZHAO

doi:10.13422/j.cnki.syfjx.20192038

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Intervention Effect of Kidney-tonifying and Essence-filling Therapy on Hippocampal Autophagy in Kidney-deficiency Alzheimer Mice

Pharmacology | 更新时间：2021-02-09

- Intervention Effect of Kidney-tonifying and Essence-filling Therapy on Hippocampal Autophagy in Kidney-deficiency Alzheimer Mice
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 20, Pages: 43-48(2019)
- 作者机构：
  
  辽宁中医药大学　中西医结合学院，沈阳　 110847
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20192038
  CLC： R2-0; R285.5; R289
- Received：23 April 2019，
  
  Published Online：05 July 2019，
  
  Published：20 October 2019
- 稿件说明：
移动端阅览
Zhong-kang ZHU, Lin ZHANG, Chun LIU, et al. Intervention Effect of Kidney-tonifying and Essence-filling Therapy on Hippocampal Autophagy in Kidney-deficiency Alzheimer Mice[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(20): 43-48.
DOI：

Zhong-kang ZHU, Lin ZHANG, Chun LIU, et al. Intervention Effect of Kidney-tonifying and Essence-filling Therapy on Hippocampal Autophagy in Kidney-deficiency Alzheimer Mice[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(20): 43-48. DOI： 10.13422/j.cnki.syfjx.20192038.

摘要

目的：

通过研究六味地黄丸对肾虚阿尔茨海默病(AD)小鼠海马神经元自噬水平的改善作用及部分机制，探讨补肾填精法对AD的部分治疗机制。

方法：

健康雄性C57-B6小鼠分为正常组，AD组，肾虚AD组，六味地黄丸组(1.08 g·kg

－1

)。正常组与AD组每日皮下注射生理盐水(15 mL·kg

－1

)，肾虚AD组和六味地黄丸组每日皮下注射氢化可的松注射液(15 mL·kg

－1

)，连续注射20 d。第21天除正常组侧脑室注射无菌生理盐水外，其余3组侧脑室注射

淀粉样蛋白25-35(A

25-35

)，6 μg/只。采用酶联免疫吸附测定(ELISA)检测各组小鼠血清皮质醇、睾酮水平；透射电镜观察海马神经元细胞形态变化；免疫荧光检测微管相关蛋白1轻链3(LC3)的表达；蛋白免疫印迹法(Western blot)检测选择性自噬接头蛋白(p62)的蛋白表达。

结果：

与正常组比较，AD组和肾虚AD组小鼠的血清皮质醇、睾酮水平显著降低(

0.01)，小鼠海马LC3表达显著减少(

0.01)，小鼠海马p62蛋白表达显著增加(

0.01)；与AD组比较，肾虚AD组小鼠的血清皮质醇、睾酮水平明显降低(

0.05，

0.01)，小鼠海马p62蛋白表达显著增加(

0.01)；与AD组和肾虚AD组比较，六味地黄丸组小鼠的血清皮质醇、睾酮水平显著升高(

0.01)，小鼠海马LC3表达显著增多(

0.01)，p62蛋白表达显著减少(

0.01)。正常组和六味地黄丸组小鼠神经元细胞内可见自噬小体，AD组和肾虚AD组小鼠神经元细胞内自噬小体鲜见。

结论：

补肾填精法能保护海马神经元细胞，增加海马神经元中LC3表达，降低p62表达水平，提高海马神经元自噬水平，对肾虚阿尔茨海默病具有一定的治疗作用。

Abstract

Objective:

To study the effect of Liuwei Dihuangwan on the improvement of autophagy level of hippocampal neurons in mice with kidney deficiency Alzheimer's disease (AD) and its partial mechanism

in order to explore part of therapeutic mechanisms of kidney-tonifying and essence-filling therapy for AD.

Method:

Healthy male C57-B6 mice were divided into control group

AD group

kidney deficiency AD group and Liuwei Dihuangwan group(1.08 g·kg

-1

). The control group and the AD group were subcutaneously injected with normal saline (15 mL·kg

-1

) daily

and the kidney deficiency AD group and the Liuwei Dihuangwan group were subcutaneously injected with hydrocortisone injection (15 mL·kg

-1

) daily for 20 consecutive days. On the 21

day

the other three groups were injected with 6 μg amyloid beta protein 25-35(A

25-35

) in the lateral ventricle

while the control group was injected with sterile saline into the lateral ventricle. The levels of serum cortisol and testosterone in each group were detected by enzyme-linked immunosorbent assay (ELISA)

the morphological changes in hippocampal neurons were observed by transmission electron microscopy

the expression of microtubule-associated protein 1 light chain 3 (LC3) was detected by immunofluorescence

and the expression of selective autophagic junction protein (p62) was detected by Western blot.

Result:

Compared with normal group

serum cortisol and testosterone levels in AD group and kidney deficiency AD group were significantly reduced (

0.01)

LC3 expression in hippocampus of mice was significantly reduced (

0.01)

and p62 protein expression in hippocampus of mice was significantly increased (

0.01). Compared with AD group

serum cortisol and testosterone levels in kidney deficiency AD group were significantly reduced (

0.05

0.01)

and p62 protein expression in hippocampus of mice was significantly increased (

0.01). Compared with AD group and kidney deficiency AD group

serum cortisol and testosterone levels of mice in Liuwei Dihuangwan group were significantly increased (

0.01)

LC3 expression in hippocampus was significantly increased (

0.01)

and p62 protein expression was significantly reduced (

0.01). Autophagosomes were observed in mouse neuron cells in normal group and Liuwei Dihuangwan group

and rarely in AD group and kidney deficiency AD group.

Conclusion:

Kidney-tonifying and essence-filling therapy can protect hippocampal neurons

increase LC3 expression in hippocampal neurons

decrease p62 expression level and increase autophagy level of hippocampal neurons. It has a certain therapeutic effect on kidney-deficiency Alzheimer's disease.

关键词

Keywords

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