小柴胡颗粒激活Nrf2通路抗TAA致大鼠急性肝损伤的机制探讨

贾为壹; 刘佳佳; 胡睿; 胡安玲; 徐尚福; 王欢; 罗果; 李晋

doi:10.13422/j.cnki.syfjx.20190840

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小柴胡颗粒激活Nrf2通路抗TAA致大鼠急性肝损伤的机制探讨

药理 | 更新时间：2021-02-09

- 小柴胡颗粒激活Nrf2通路抗TAA致大鼠急性肝损伤的机制探讨
- Therapeutic Mechanism of Xiaochaihu Granule on Acute Liver Injury Induced by Thioacetamide in Rats Through Nrf2 Pathway
- 中国实验方剂学杂志 2019年25卷第8期页码：54-59
- 作者机构：
  
  遵义医科大学　医学与生物学研究中心，基础药理教育部重点实验室暨特色民族药教育部国际合作联合实验室　贵州省普通高等学校脑科学特色重点实验室，贵州　遵义　 563003
- 作者简介：
  
  贾为壹，在读硕士，从事中西医结合治疗肝脏相关疾病研究，E-mail: vickyj1@163.com
  李晋，副教授，硕士生导师，从事中西医结合治疗消化系统疾病研究，E-mail: lijin010502@163.com
- 基金信息：
  
  国家自然科学基金项目(81360661)
- DOI：10.13422/j.cnki.syfjx.20190840
  中图分类号： R289;R575.1;944.2+7
- 收稿日期：2018-10-17，
  
  网络出版日期：2019-01-04，
  
  纸质出版日期：2019-04-20
- 稿件说明：
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贾为壹, 刘佳佳, 胡睿, 等. 小柴胡颗粒激活Nrf2通路抗TAA致大鼠急性肝损伤的机制探讨[J]. 中国实验方剂学杂志, 2019,25(8):54-59.

Wei-yi JIA, Jia-jia LIU, Rui HU, et al. Therapeutic Mechanism of Xiaochaihu Granule on Acute Liver Injury Induced by Thioacetamide in Rats Through Nrf2 Pathway[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(8): 54-59.
贾为壹, 刘佳佳, 胡睿, 等. 小柴胡颗粒激活Nrf2通路抗TAA致大鼠急性肝损伤的机制探讨[J]. 中国实验方剂学杂志, 2019,25(8):54-59. DOI： 10.13422/j.cnki.syfjx.20190840.

Wei-yi JIA, Jia-jia LIU, Rui HU, et al. Therapeutic Mechanism of Xiaochaihu Granule on Acute Liver Injury Induced by Thioacetamide in Rats Through Nrf2 Pathway[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(8): 54-59. DOI： 10.13422/j.cnki.syfjx.20190840.

摘要

目的：

观察小柴胡颗粒对硫代乙酰胺(TAA)致大鼠急性肝损伤(ALI)的治疗作用，探讨核转录因子(NF)-E2相关因子2(Nrf2)抗氧化损伤通路在其中的作用。

方法：

SD大鼠随机分为空白组，模型组，小柴胡颗粒低、中、高剂量组(1

4 g·kg

-1

)，大鼠给予250 mg·kg

-1

TAA腹腔注射2 d制备肝损伤模型，从第3天各组分别给予等量双蒸水和不同剂量小柴胡颗粒，灌胃2周。末次给药后24 h取材，肝组织行苏木素-伊红(HE)染色；比色法测定血清中丙氨酸氨基转移酶(ALT)，天门冬氨酸氨基转移酶(AST)活性及组织中总超氧化物歧化酶(T-SOD)，丙二醛(MDA)水平；实时荧光定量PCR(Real-time PCR)检测Nrf2通路相关mRNA表达；蛋白免疫印迹法(Western blot)检测Nrf2通路相关蛋白的表达。

结果：

与空白组比较，模型组ALT

AST

MDA水平显著增高

T-SOD活性明显降低(

0.05，

0.01)，病理学呈现大片炎性浸润表现，肝脏组织Nrf2，Kelch样环氧氯丙烷相关蛋白1(Keap1)，醌氧化还原酶1(NQO1)，血红素加氧酶-1(HO-1)，谷氨酸半胱氨酸合成酶催化亚基(GCLC)，谷氨酸半胱氨酸合成酶调节亚基(GCLM)mRNA及蛋白表达水平均明显下降(

0.05，

0.01)；与模型组比较，小柴胡颗粒各剂量组ALT

AST

MDA水平均明显降低

T-SOD活性明显升高(

0.05，

0.01)，病理结果示大鼠肝脏病变范围与严重程度均有不同程度改善，肝脏组织Nrf2，Keap1

NQO1

HO-1

GCLC

GCLM的mRNA及蛋白表达水平均明显升高(

0.05，

0.01)。

结论：

小柴胡颗粒可能通过上调Nrf2信号通路上下游分子的表达，对TAA致急性肝损伤大鼠起到治疗作用。

Abstract

Objective:

To observe the therapeutic effect of Xiaochaihu granule on acute liver injury (ALI) induced by thioacetamide (TAA) in rats

and to explore the role of transcription factor (NF)-E2 related factor 2 (Nrf2) in the pathway of oxidative damage.

Method:

SD rats were randomly divided into control group

model group

Xiaochaihu granule low

middle and high dose groups (1

4 g·kg

-1

). 250 mg·kg

-1

TAA was given to the rats by

. administration for 2 days to prepare the liver injury model

and from the 3

day

same amount of double distilled water or different doses of Xiaochaihu granule was given to corresponding groups by

. administration for 2 weeks. 24 hours after the last administration

liver tissues were taken and stained with hematoxylin-eosin(HE) staining. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were measured by colorimetry. The activities of total superoxide dismutase (T-SOD) and malondialdehyde (MDA) in liver tissues were measured by colorimetry. Real-time PCR was used to detect Nrf2 pathway related mRNA expression.

Result:

As compared with control group

ALT

AST and MDA in model group were significantly increased

while T-SOD was significantly decreased (

0.05

0.01)

and pathological features showed large inflammatory infiltration

both mRNA and protein expression levels of Nrf2

Kelch-like ECH-associated protein 1(Keap1)

quinone oxidoreductase(NQO1)

heme oxygenase-1(HO-1)

glutamate-cysteine ligase catalytic subunit(GCLC) and glutamate-cysteine ligase modifier subunit(GCLM) were decreased to different degrees (

0.01). As compared with model group

ALT

AST

MDA levels in Xiaochaihu granule treatment groups were decreased

while T-SOD was increased significantly (

0.05

0.01). Pathological results showed that the range and severity of liver lesions in rats were improved to varying degrees after treatment

both mRNA and protein expression levels of Nrf2

Keap1

NQO1

HO-1

GCLC and GCLM were increased to varying degrees (

0.05

0.01).

Conclusion:

Xiaochaihu granule may play a therapeutic role in TAA induced ALI in rats by up-regulating the expression of downstream molecules in Nrf2 signaling pathway.

关键词

Keywords

references

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