基于Nrf2/Keap1/p62信号通路探讨百合固金汤对LPS诱导的小鼠急性肺损伤的保护作用

张敏; 许成辰; 徐婷贞; 董春鸽

doi:10.13422/j.cnki.syfjx.20191540

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基于Nrf2/Keap1/p62信号通路探讨百合固金汤对LPS诱导的小鼠急性肺损伤的保护作用

经典名方 | 更新时间：2021-02-09

- 基于Nrf2/Keap1/p62信号通路探讨百合固金汤对LPS诱导的小鼠急性肺损伤的保护作用
- Effect of Baihe Gujin Tang on LPS-induced Acute Lung Injury in Mice Based on Nrf2/Keap1/p62 Signaling Pathway
- 中国实验方剂学杂志 2019年25卷第15期页码：77-82
- 作者机构：
  
  浙江中医药大学　附属第一医院，杭州　 310000
- 作者简介：
  
  张敏，硕士，住院医师，从事药理与毒理学研究，E-mail：zjzyyzm@163.com
  董春鸽，硕士，副主任医师，从事呼吸系统相关疾病研究，E-mail：634923029@qq.com
- 基金信息：
  
  浙江省中医药科技计划项目(2016ZA067)
- DOI：10.13422/j.cnki.syfjx.20191540
  中图分类号： R2-0; R285; R289; R318.13
- 收稿日期：2019-01-17，
  
  网络出版日期：2019-04-29，
  
  纸质出版日期：2019-08-05
- 稿件说明：
移动端阅览
张敏, 许成辰, 徐婷贞, 等. 基于Nrf2/Keap1/p62信号通路探讨百合固金汤对LPS诱导的小鼠急性肺损伤的保护作用[J]. 中国实验方剂学杂志, 2019,25(15):77-82.

Min ZHANG, Cheng-chen XU, Ting-zhen XU, et al. Effect of Baihe Gujin Tang on LPS-induced Acute Lung Injury in Mice Based on Nrf2/Keap1/p62 Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(15): 77-82.
张敏, 许成辰, 徐婷贞, 等. 基于Nrf2/Keap1/p62信号通路探讨百合固金汤对LPS诱导的小鼠急性肺损伤的保护作用[J]. 中国实验方剂学杂志, 2019,25(15):77-82. DOI： 10.13422/j.cnki.syfjx.20191540.

Min ZHANG, Cheng-chen XU, Ting-zhen XU, et al. Effect of Baihe Gujin Tang on LPS-induced Acute Lung Injury in Mice Based on Nrf2/Keap1/p62 Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(15): 77-82. DOI： 10.13422/j.cnki.syfjx.20191540.

摘要

目的：

探究百合固金汤对脂多糖诱导的急性肺损伤(LPS-ALI)小鼠的预防作用及其机制。

方法：

将KM小鼠随机分为5组：空白组，模型组，地塞米松组(0.002 g·kg

－1

)，百合固金汤组(0.417，1.25 g·kg

－1

)。除空白组外，其余各组采用LPS诱导小鼠ALI模型。给药组分别在造模前的第1～7天连续灌胃给药7 d。在造模后6 h取小鼠肺组织，测定左肺湿/干质量比(W/D)；收集小鼠血清检测超氧化物歧化酶(SOD)，丙二醛(MDA)，活性氧(ROS)，一氧化氮(NO)的水平；应用苏木素-伊红(HE)染色方法观察肺组织病理改变；蛋白免疫印迹法(Western blot)检测肺组织中核转录因子E2相关因子2(Nrf2)，Kelch样环氧氯丙胺相关蛋白1(Keap1)，p62以及自噬相关蛋白(LC3)Ⅱ蛋白的表达水平。

结果：

与空白组比较，模型组小鼠W/D显著升高(

0.01)，血清中MDA水平显著性升高(

0.01)，SOD显著降低(

0.01)，肺组织病变严重，并显著降低肺组织Nrf2，Keap1，升高p62，LC3Ⅱ的蛋白表达(

0.01)。与模型组比较，地塞米松组、百合固金汤1.25 g·kg

－1

组W/D 显著降低(

0.01)；百合固金汤1.25 g·kg

－1

组及地塞米松组，能够明显抑制血清中MDA的水平(

0.05，

0.01)，升高SOD(

0.05，

0.01)水平，地塞米松组及百合固金汤各剂量组对小鼠肺组织病变较模型组有不同程度的改善，且显著性升高肺组织Nrf2，Keap1，降低p62及LC3Ⅱ的蛋白表达(

0.05，

0.01)，但百合固金汤各剂量组对LC3Ⅱ无影响。

结论：

百合固金汤对LPS-ALI小鼠具有明显的预防作用，其机制可能与调控Nrf2/Keap1/自噬反馈回路有关。

Abstract

Objective:

To investigate the protective effect and mechanism of Baihe Gujin Tang on lipopolysaccharide induced acute lung injury (LPS-ALI).

Method:

KM mice were randomly divided into 5 groups: blank control group

model group

dexamethasone 0.002 g·kg

-1

group

Baihe Gujin Tang (0.417

1.25 g·kg

-1

) group. Except for the blank control group

the other groups were given LPS to induce the mouse ALI model. Except for the blank control group and the model group

the other groups were continuously given intragastric administration for 7 days on the 1

to 7

days before modeling. The lung tissue of the mice was taken 6 h after modeling

and the wet/dry mass ratio (W/D) of the left lung was measured. The serum levels of superoxide dismutase(SOD)

malondialdehyde (MDA)

reactive oxygen species (ROS)and nitric oxide (NO) were detected in the mice. Thepathological changes of the lung tissues were observed by hematoxylin-eosin(HE) staining. The expression levels of nuclear factor E2 related factor 2(Nrf2)

Kelch-likeECH-associated protein 1 (Keap1)

p62 and autophagy associated proteinsLC3Ⅱ proteins in the lung tissues were detected by Western blot.

Result:

Compared with the blank control group

the W/D of the model group was significantly increased (

0.01)

the MDA level in the serum was significantly increased (

0.01)

and the SOD was significantly decreased (

0.01)

lung tissue lesions were severe

and significantly reduced the expression of lung tissue Nrf2

Keap1

but increased p62

LC3Ⅱ protein expression (

0.01). Compared with model group

the W/D of dexamethasone group and Baihe Gujin Tang 1.25 g·kg

-1

group was significantly lower (

0.01). Baihe Gujin Tang 1.25 g·kg

-1

group and dexamethasone group were able to significantly inhibited MDA levels in serum (

0.05

0.01)

improved the expression of SOD (

0.05

0.01)

dexamethasone group and Baihe Gujin Tang group have different extents of improvement the pathological changes of the lung tissues the pathological changes of the lung tissues

and the lung tissue Nrf2 and Keap1 were significantly increased

and the protein expression of p62

LC3Ⅱwas decreased (

0.05

0.01)

each dose group of Baihe Gujin Tang had no effect on LC3B.

Conclusion:

Baihe Gujin Tang has obvious protective effect on LPS-ALI mice

and its mechanism may be related to the regulation of Nrf2/Keap1/autophagy feedback loop.

关键词

Keywords

references

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