加味香砂六君子汤调控高脂血症小鼠apoA-Ⅰ改善内质网应激的作用机制研究

张琦; 隋国媛; 宋囡; 王杰; 刘玉; 蔡浩冉; 贾连群

doi:10.3969/j.issn.1006-2157.2024.09.008

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加味香砂六君子汤调控高脂血症小鼠apoA-Ⅰ改善内质网应激的作用机制研究

实验研究 | 更新时间：2024-10-15

- 加味香砂六君子汤调控高脂血症小鼠apoA-Ⅰ改善内质网应激的作用机制研究
- The mechanism of modified Xiangsha Liujunzi Decoction in regulating apoA-I and improving endoplasmic reticulum stress in hyperlipidemic mice
- 北京中医药大学学报 2024年47卷第9期页码：1236-1246
- 作者机构：
  
  1.辽宁中医药大学中医药创新工程技术中心脏象理论及应用教育部重点实验室　沈阳　110847
  2.辽宁中医药大学中西医结合学院
- 作者简介：
  
  张琦，女，在读博士生
  #贾连群，女，博士，教授，博士生导师，主要研究方向：中西医结合防治心血管疾病，E-mail：jlq－8@163.com
- 基金信息：
  
  国家自然科学基金项目(82074145;81974548;82374423)
- DOI：10.3969/j.issn.1006-2157.2024.09.008
  中图分类号： R285.5
- 收稿：2024-03-22，
  
  网络首发：2024-08-15，
  
  纸质出版：2024-09-30
- 稿件说明：
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张琦, 隋国媛, 宋囡, 等. 加味香砂六君子汤调控高脂血症小鼠apoA-Ⅰ改善内质网应激的作用机制研究[J]. 北京中医药大学学报, 2024,47(9):1236-1246.

ZHANG Qi, SUI Guoyuan, SONG Nan, et al. The mechanism of modified Xiangsha Liujunzi Decoction in regulating apoA-I and improving endoplasmic reticulum stress in hyperlipidemic mice[J]. Journal of Beijing University of Traditional Chinese Medicine, 2024, 47(9): 1236-1246. DOI: 10.3969/j.issn.1006-2157.2024.09.008.
张琦, 隋国媛, 宋囡, 等. 加味香砂六君子汤调控高脂血症小鼠apoA-Ⅰ改善内质网应激的作用机制研究[J]. 北京中医药大学学报, 2024,47(9):1236-1246. DOI： 10.3969/j.issn.1006-2157.2024.09.008.

ZHANG Qi, SUI Guoyuan, SONG Nan, et al. The mechanism of modified Xiangsha Liujunzi Decoction in regulating apoA-I and improving endoplasmic reticulum stress in hyperlipidemic mice[J]. Journal of Beijing University of Traditional Chinese Medicine, 2024, 47(9): 1236-1246. DOI: 10.3969/j.issn.1006-2157.2024.09.008. DOI：

摘要

目的

探讨加味香砂六君子汤调控小鼠载脂蛋白A－Ⅰ(apoA-Ⅰ)改善内质网应激调节糖脂代谢防治血脂异常的机制。

方法

将野生型C57BL/6J小鼠根据随机数字表法分为野生组、野生高脂组、野生中药干预组，将apoA-Ⅰ基因敲除小鼠根据随机数字表法分为敲除组、敲除高脂组、敲除中药干预组，每组10只。除野生组和敲除组正常饮食喂养外，各组小鼠采用D12492高脂饲料进行高脂饮食喂饲，建立高脂血症小鼠模型。于第9周开始野生中药干预组和敲除中药干预组小鼠每天给予加味香砂六君子汤(23.66 g/kg)灌胃给药，4周后，收集小鼠血清及肝脏组织用于后续检测。全自动生化分析仪检测血脂水平；ELISA法检测血清空腹血糖(FBG)及胰岛素(INS)水平，计算胰岛素抵抗(HOMA-IR)指数；HE染色观察肝脏病理变化；油红O染色观察肝脏脂质沉积情况；微板法检测肝脏组织甘油三酯(TG)水平；实时荧光PCR法检测肝脏组织apoA-Ⅰ、葡萄糖调节蛋白78(GRP78)、固醇调节元件结合蛋白－1c(SREBP-1c)、乙酰辅酶A羧化酶1(ACC1)、脂肪酸合酶(FASN)mRNA表达，WES全自动蛋白表达分析系统检测肝脏组织apoA-Ⅰ、GRP78、肌醇需求酶1(IRE1)、磷酸化IREI(p-IRE1)、c-Jun氨基末端激酶(JNK)、磷酸化JNK(p-JNK)、胰岛素受体底物1(IRS1)、磷酸化IRS1(p-IRS1)、蛋白激酶B(Akt)、p-Akt、SREBP-1c、ACC1、FASN蛋白表达。

结果

野生高脂组相较于野生组血清中血脂、FBG、INS水平及HOMA-IR指数升高(

0.05)；肝脏组织中橘红色脂滴增多，脂肪空泡明显，TG水平增加，apoA-Ⅰ mRNA及蛋白表达降低，GRP78、SREBP-1c、ACC1、FASN mRNA表达升高，GRP78、SREBP-1c、ACC1、FASN蛋白水平及IRE1、JNK、IRS1、Akt磷酸化程度升高(

0.05)。野生中药干预组与野生高脂组比较，血清中TC、TG、HDL-C、LDL-C、FBG、INS水平及HOMA-IR指数降低(

0.05)；肝脏组织中橘红色脂滴明显减少，脂肪空泡化减轻，TG水平降低，apoA-Ⅰ mRNA及蛋白表达升高，GRP78、SREBP-1c、ACC1、FASN mRNA表达水平降低，GRP78、SREBP-1c、ACC1、FASN蛋白水平及IRE1、JNK、IRS1、Akt磷酸化程度降低(

0.05)。敲除高脂组相较于野生高脂组，血清中TG、LDL-C、FBG水平及HOMA-IR指数升高，HDL-C水平降低(

0.05)；肝脏组织弥漫性橘红色脂滴，脂肪空泡明显增多，TG水平升高，GRP78、SREBP-1c、ACC1、FASN mRNA表达水平升高，SREBP-1c、ACC1蛋白水平及IRE1、JNK、IRS1、Akt磷酸化程度升高(

0.05)。敲除中药干预组相较于野生中药干预组，血清中TG、LDL-C、FBG、INS水平及HOMA-IR指数升高，HDL-C

水平降低(

0.05)；肝脏组织橘红色脂滴沉积情况改善，TG水平降低，GRP78、SREBP-1c、ACC1、FASN mRNA表达水平升高，GRP78、SREBP-1c、ACC1、FASN蛋白水平及IRE1、JNK、IRS1、Akt磷酸化程度升高(

0.05)。

结论

加味香砂六君子汤通过调控apoA-Ⅰ缓解内质网应激，进而改善肝脏糖脂代谢紊乱。

Abstract

Objective

To explore the mechanism of modified

Xiangsha Liujunzi

Decoction in regulating apolipoproteinA-I (apoA-I)

improving endoplasmic reticulum stress

regulating glucose and lipid metabolism

and preventing and treating dyslipidemia in mice.

Methods

Wild-type (WT) C57BL/6J mice were randomly divided into the WT

WT + high-fat diet(HFD)

and WT + HFD +

Xiangsha Liujunzi

Decoction(XSLJZ) groups according to the random number table method. ApoA-Ⅰ

-/-

mice were randomly divided into the apoA-Ⅰ

-/-

apoA-Ⅰ

-/-

+ HFD

and apoA-Ⅰ

-/-

+ HFD + XSLJZ groups (

=10) according to the random number table method. D12492 was used for HFD feeding to establish a hyperlipidemic mouse model. Modified XSLJZ (23.66 g/kg) was administered daily by gavage from the ninth week. Serum and liver tissue were collected for testing after 4 weeks. An automatic biochemical analyzer was used to detect blood lipid levels; an enzyme-linked immunosorbent assay was used to detect serum fasting blood glucose (FBG) and insulin (INS) levels

and the INS resistance index (HOMA-IR) was calculated. Hematoxylin and eosin staining was used to observe the pathological changes in the liver. Oil red O staining was used to observe the lipid deposition in the liver. TG levels in liver tissue were detected using the microplate method. Real-time PCR was used to detect apoA-Ⅰ

glucose-regulated proteins (GRP78)

sterol regulatory element binding protein-1c (SREBP-1c)

acetyl CoA carboxylase 1 (ACC1)

and fatty acid synthase (FASN) mRNA expression levels in liver tissue. The WES fully automated prote

in expression analysis system was used to detect apoA-Ⅰ

GRP78

inositol-requiring enzyme 1 (IRE1)

p-IRE1

c-Jun N-terminal kinase (JNK)

p-JNK

insulin receptor substrate (IRS1)

p-IRS1

protein kinase B (Akt)

p-Akt

SREBP-1c

ACC1

and FASN protein expression levels in liver tissue.

Results

Compared to the WT group

the WT + HFD group showed a significant increase in serum lipids

FBG

INS levels

and the HOMA-IR index (

0.05). The orange-red lipid droplets in liver tissue increased

fat vacuoles were apparent

and TG levels were significantly increased. ApoA-Ⅰ mRNA and protein expression levels were significantly reduced

whereas GRP78

SREBP-1c

ACC1

and FASN mRNA expression levels were increased

GRP78

SREBP-1c

ACC1

and FASN protein levels and the IRE1

JNK

IRS1

and Akt phosphorylation degree were increased (

0.05). The serum TG

HDL-C

LDL-C

FBG

and INS levels and the HOMA-IR index in the WT + HFD group were significantly reduced after administering modified XSLJZ (

0.05). The orange-red lipid droplets in liver tissue were significantly reduced

fat vacuolization was alleviated

and TG levels were significantly reduced

ApoA-Ⅰ mRNA and protein expression levels were significantly increased

whereas GRP78

SREBP-1c

ACC1

and FASN mRNA expression levels were reduced

GRP78

SREBP-1c

ACC1

and FASN protein expression levels and the IRE1

JNK

IRS1

and Akt phosphorylation degree were reduced (

0.05). Compared to the WT + HFD group

the TG

LDL-C

and FBG levels and HOMA-IR index in the serum of the apoA-Ⅰ

-/-

+ HFD group were significantly increased

whereas the HDL-C levels were significantly decreased (

0.05). Diffuse orange-red lipid droplets in liver tissue and a significant increase in fat vacuoles were observed. Furthermore

TG levels were significantly increased

SREBP-1c

ACC1

FASN mRNA

SREBP-1c

and ACC1 protein expression levels and

IRE1

JNK

IRS1

and Akt phosphorylation levels were significantly increased (

0.05). Compared to the WT + HFD + XSLJZ group

the apoA-Ⅰ

-/-

+ HFD + XSLJZ group showed a significant increase in serum TG

LDL-C

FBG

and INS levels

and the HOMA-IR index

whereas HDL-C levels decreased significantly (

0.05). The deposition of orange-red lipid droplets in liver tissue improved

and TG levels significantly decreased

GRP78

SREBP-1c

ACC1

and FASN mRNA expression levels

GRP78

SREBP-1c

and ACC1 protein levels

and IRE1

JNK

IRS1

and Akt phosphorylation levels increased (

0.05).

Conclusion

Modified XSLJZ improves liver glucose and lipid metabolism disorder by regulating apoA-Ⅰ to alleviate endoplasmic reticulum stress.

关键词

Keywords

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