补肾填精法对肾虚阿尔茨海默小鼠海马自噬的干预作用

朱仲康; 张林; 柳春; 王旭; 刘慧慧; 赵丹玉

doi:10.13422/j.cnki.syfjx.20192038

您当前的位置：

首页 >

文章列表页 >

补肾填精法对肾虚阿尔茨海默小鼠海马自噬的干预作用

药理 | 更新时间：2021-02-09

- 补肾填精法对肾虚阿尔茨海默小鼠海马自噬的干预作用
- Intervention Effect of Kidney-tonifying and Essence-filling Therapy on Hippocampal Autophagy in Kidney-deficiency Alzheimer Mice
- 中国实验方剂学杂志 2019年25卷第20期页码：43-48
- 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：
  
  朱仲康，在读硕士，从事中医药防治代谢性疾病分子机制的研究，E-mail：zzk1223@foxmail.com
  赵丹玉，博士，教授，从事中医药防治代谢性疾病分子机制的研究，E-mail：danyu1978@163.com
- 基金信息：
  
  国家自然科学基金项目(81803854);辽宁省自然科学基金项目(20121100)
- DOI：10.13422/j.cnki.syfjx.20192038
  中图分类号： R2-0; R285.5; R289
- 收稿日期：2019-04-23，
  
  网络出版日期：2019-07-05，
  
  纸质出版日期：2019-10-20
- 稿件说明：
移动端阅览
朱仲康, 张林, 柳春, 等. 补肾填精法对肾虚阿尔茨海默小鼠海马自噬的干预作用[J]. 中国实验方剂学杂志, 2019,25(20):43-48.

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.
朱仲康, 张林, 柳春, 等. 补肾填精法对肾虚阿尔茨海默小鼠海马自噬的干预作用[J]. 中国实验方剂学杂志, 2019,25(20):43-48. DOI： 10.13422/j.cnki.syfjx.20192038.

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

references

Thal D R , Fndrich M . Protein aggregation in Alzheimer' s disease: Aβ and τ and their potential roles in the pathogenesis of AD [J]. Acta Neuropathol , 2015 , 129 ( 2 ): 163 - 165 .

Jo C , Gundemir S , Pritchard S , et al . Nrf2 reduces levels of phosphorylated tau protein by inducing autophagy adaptor protein NDP52 [J]. Nat Commun , 2014 , 5 : 3496 .

Hensley K , Harris W M E . Redox regulation of autophagy in healthy brain and neurodegeneration [J]. Neurobiol Dis , 2015 , 84 : 50 - 59 .

LI L , ZHANG X , Le W . Autophagy dysfunction in Alzheimer’s disease [J]. Neurodegener Dis , 2010 , 7 ( 4 ): 265 - 271 .

Madeo F , Eisenberg T , Kroemer G . Autophagy for the avoidance of neurodegeneration [J]. Genes Dev , 200 , 23 ( 19 ): 2253 - 2259 .

王苏雷．冬凌草甲素改善阿尔茨海默病模型小鼠认知功能及机制研究 [D]．南京：南京中医药大学， 2016 ．

任晨斌，伍大华．国医大师刘祖贻用温肾活血法治疗阿尔茨海默病经验 [J]．中医药导报， 2016 ， 22 ( 16 )： 14-15，25 ．

韦云，刘剑刚，李浩，等．从中医阴阳理论探讨神经细胞自噬现象对阿尔茨海默病的影响 [J]．中医杂志， 2013 ， 54 ( 13 )： 1085 - 1087 ．

李欢，苗明三．自噬及中医药调控自噬在治疗和防治疾病中的研究进展 [J]．中国实验方剂学杂志， 2018 ， 24 ( 20 )： 200 - 207 ．

章梦琦，滕燕，王文，等．自噬在阿尔茨海默病中的作用及中药的干预作用 [J]．中国实验方剂学杂志， 2013 ， 19 ( 17 )： 355 - 359 ．

罗俊，张科楠，肖帅，等．镇心省睡益智方及其精油对AD模型小鼠学习记忆的影响 [J]．中国实验方剂学杂志， 2019 ， 25 ( 9 )： 74 - 81 ．

王璐，孙丽娥，康湘萍，等．补肾益气方药左归丸和益气聪明汤对老年大鼠学习记忆能力及相关基因表达的影响 [J]．中国实验方剂学杂志， 2019 ， 25 ( 6 )： 15 - 22 ．

崔勇，王艳杰，赵丹玉，等．六味地黄丸对APP/PS1小鼠学习记忆能力及胆碱能系统的影响 [J]．中国实验方剂学杂志， 2016 ， 22 ( 3 )： 148 - 151 ．

周岚．老年痴呆的中医辨证分型特点及其与APOE基因多态性相关性的初探 [D]．武汉：湖北中医学院， 2004 ．

陈奇．中药药理实验方法 [M]．北京：人民卫生出版社， 1994 ： 176 - 179 ．

黄贵华，纪云西，吴大力，等．细胞自噬与中医气虚痰瘀关系探讨 [J]．中医杂志， 2011 ， 52 ( 20 )： 1717 - 1719 ．

马妍，闫晨，张连城，等．老年性痴呆肾虚痰瘀发病机制中细胞自噬研究进展 [J]．中国老年学杂志， 2017 ， 37 ( 10 )： 2570 - 2572 ．

李长禄，王红梅，王立为．肾虚型老年痴呆大鼠模型的复制 [J]．安徽中医学院学报， 2013 ， 32 ( 3 )： 62 - 67 ．

王秀凤，李静，张磊，等．基于因子分析的肾阳虚证动物模型垂体―靶腺轴关键指标的评价 [J]．中国中西医结合杂志， 2013 ， 33 ( 6 )： 825 - 829 ．

宋彩梅，王红梅，刘新民，等．肾虚型老年痴呆动物模型的建立 [J]．现代中西医结合杂志， 2011 ， 20 ( 22 )： 2744 - 2748 ．

杜欣芸，李常青，陈滨，等．叶下珠复方Ⅱ号对肝癌Huh7细胞增殖、凋亡与自噬的影响 [J]．中国实验方剂学杂志， 2019 ， 25 ( 2 )： 48 - 54 ．

Kabeya Y , Mizushima N , Ueno T , et al . LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing [J]. EMBO J , doi: 10.1093/emboj/19.21.5720 http://doi.org/10.1093/emboj/19.21.5720 .

Park S , Choi S G , Yoo S M , et al . Choline dehydrogenase interacts with SQSTM1/p62 to recruit LC3 and stimulate mitophagy [J]. Autophagy , 2014 , 10 ( 11 ): 1906 - 1920 .

游宇，刘玉晖，廖旺娣．参苓白术散抗炎症性肠病作用与调节肠上皮细胞自噬的关系探讨 [J]．中国实验方剂学杂志， 2019 ， 25 ( 15 )： 51 - 56 ．

Komatsu M , Kurokawa H , Waguri S , et al . The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1 [J]. Nat Cell Biol , 2010 , 12 ( 3 ): 399 - 403 .

WANG X , LI L , NIN X , et al . mTOR enhances foam cell formation by suppressing the autophagy pathway [J]. DNA Cell Biol , 2014 , 33 ( 4 ): 198 - 204 .

Levine B , Kroemer G . Autophagy in the pathogenesis of disease [J]. Cell , 2008 , 132 ( 1 ): 27 - 42 .

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

六味地黄丸通过PI3K/Akt/FoxO3a通路调控自噬对SAMP8小鼠记忆功能的影响

黄精丸对学习记忆障碍小鼠海马神经元自噬的影响

六味地黄丸对秀丽隐杆线虫AD模型线粒体损伤的保护作用

地黄饮子改善AD小鼠脑星形胶质细胞能量代谢障碍及自噬损伤的作用机制