肝豆汤对Wilson病模型TX小鼠肝细胞内铜代谢通路的分子调控机制

董健健; 韩咏竹; 程楠

doi:10.13422/j.cnki.syfjx.2016240128

您当前的位置：

首页 >

文章列表页 >

肝豆汤对Wilson病模型TX小鼠肝细胞内铜代谢通路的分子调控机制

药理 | 更新时间：2024-01-04

- 肝豆汤对Wilson病模型TX小鼠肝细胞内铜代谢通路的分子调控机制
- Molecular Regulatory Mechanism of Gandou Decoction on Copper Metabolism Pathway in Hepatocytes of Wilson's Disease Model-TX Mice
- 中国实验方剂学杂志 2016年22卷第24期页码：128-133
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金项目（81473535，81173212）
- DOI：10.13422/j.cnki.syfjx.2016240128
  中图分类号： R-332;R277.7
- 纸质出版日期：2016
- 稿件说明：
移动端阅览
董健健, 韩咏竹, 程楠. 肝豆汤对Wilson病模型TX小鼠肝细胞内铜代谢通路的分子调控机制[J]. 中国实验方剂学杂志, 2016,22(24):128-133.

DONG Jian-jian, HAN Yong-zhu, CHENG Nan. Molecular Regulatory Mechanism of Gandou Decoction on Copper Metabolism Pathway in Hepatocytes of Wilson's Disease Model-TX Mice[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(24): 128-133.
董健健, 韩咏竹, 程楠. 肝豆汤对Wilson病模型TX小鼠肝细胞内铜代谢通路的分子调控机制[J]. 中国实验方剂学杂志, 2016,22(24):128-133. DOI： 10.13422/j.cnki.syfjx.2016240128.

DONG Jian-jian, HAN Yong-zhu, CHENG Nan. Molecular Regulatory Mechanism of Gandou Decoction on Copper Metabolism Pathway in Hepatocytes of Wilson's Disease Model-TX Mice[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(24): 128-133. DOI： 10.13422/j.cnki.syfjx.2016240128.

摘要

目的：探讨肝豆汤调控毒牛奶（TX）小鼠肝细胞内铜转运通路的分子靶点并观察其相应的调控机制。方法： 20只家兔适应性喂养3 d后，随机分为空白组（生理盐水）和肝豆汤组（4.5 g·kg-1·d-1），每日2次ig，连续10 d，制得含药血清；采用2步灌注法分离提取DL，TX小鼠肝细胞，采用原子吸收法检测加含不同百分含量（5%，10%，15%，20%）含肝豆汤兔血清作用24 h后，对TX小鼠肝细胞内微量元素的影响，另设空白组；蛋白质免疫印迹（Western blot）法检测经含肝豆汤兔血清作用后腺嘌呤核苷三磷酸7b（adenosine triphosphate7b，ATP7b），抗氧化蛋白1（anti-oxidant1，ATOX1），铜转运体1（copper transporter1，CTR1），金属巯蛋白（metallothionein，MT），超氧化剂物歧化酶铜伴侣蛋白（copper chaperone of superoxide dismutase，CCS），细胞色素氧化酶17（cytochrome-coxidase17，COX17）蛋白的表达。结果：与空白组比较，模型组TX小鼠肝细胞内铜含量升高，锌含量降低，铁含量升高，ATP7b，ATOX1，CCS，COX17蛋白表达明显降低，CTR1，MT蛋白表达显著升高（P<0.01）；与模型组比较，含肝豆汤兔血清可显著降低TX小鼠肝细胞内铜含量，增加锌含量，降低铁含量（P<0.05，P<0.01）；Western blot检测结果显示，含肝豆汤兔血清可明显增加ATP7b，ATOX1，CCS，COX17蛋白表达（P<0.05，P<0.01），降低CTR1，MT蛋白表达（P<0.01）。结论：肝豆汤可能是通过调控细胞内铜代谢相关蛋白，上调ATP7b，ATOX1，CCS，COX17蛋白表达，抑制CTR1，MT表达，从而发挥细胞内排铜作用。肝豆汤可通过多靶点、多途径调控铜代谢通路达到降低肝细胞内铜含量的治疗效果。

Abstract

Objective: To detect the molecular target of copper transport pathway in hepatocytes of TX mice with Gandou decoction (GDD) treatment

and explore its corresponding regulatory metabolism. Method: Totally 20 rabbits were randomly divided into normal group (normal saline) and GDD group (4.5 g·kg-1·d-1) after adaptive feeding for 3 days. Treatment was given by ig

bid

for 10 days to obtain drug serum. Hepatocytes of DL and TX mice were isolated and extracted by 2 step perfusion method; atomic absorption method was used to detect the concentration of microelement in the hepatocytes of TX mice after treatment with rabbit serum containing different concentrations of GDD; and another blank group was set up. Western blot was used to detect the expression levels of adenosine triphosphate7b (ATP7b)

anti-oxidant1 (ATOX1)

copper transporter1 (CTR1)

metallothionein (MT)

copper chaperone of superoxide dismutase (CCS)

and cytochrome-coxidase17 (COX17) in the hepatocytes of TX mice after being treated with rabbit serum containing GDD. Result: As compared with blank group

the contents of copper and iron were increased; the content of zinc was reduced; the protein expression levels of ATP7b

ATOX1 CCS

COX17 were significantly reduced

and the protein expression levels of MT

CTR1 were significantly increased in hepatocytes of model group TX mice (P<0.01). As compared with the model group

GDD could reduce the contents of copper and iron in the hepatocytes of TX mice

and increase the content of zinc in a dose-dependent manner(P<0.05

P<0.01). Western blot results indicated that GDD could significantly increases the protein expression levels of ATP7b

ATOX1 CCS

COX17 (P<0.05

P<0.01)and decrease the protein expression levels of MT

CTR1 in the hepatocytes of TX mice (P<0.01). Conclusion: GDD may increases the copper excretion in hepatocytes of TX mice by up-regulating the protein expression levels of ATP7b

ATOX1

CCS

COX17 and down-regulating the expression levels of CTR1 and MT. GDD could reduce copper contents in hepatocytes through a multi-target and multi-channel regulation of copper metabolism pathway.

关键词

Keywords

references

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

肝豆汤对Wilson病模型TX小鼠脾脏免疫功能调节的作用

肝豆汤联合言语训练治疗湿热内蕴型Wilson病构音障碍的临床疗效

肝豆汤治疗湿热内蕴型Wilson病伴快动眼期睡眠行为障碍的临床疗效

肝豆汤对Wilson病模型高铜诱导的SH-SY5Y细胞自噬效应的影响及其作用机制

基于Cer通路探讨肝豆汤对Wilson病模型高铜诱导海马神经元细胞损伤的保护作用