Effect and Mechanism of Celastrol on Endoplasmic Reticulum Stress in Hepatic L02 Cells of Nonalcoholic Fatty Liver Disease

Xia JIANG; Hui-ling JIANG; Fan YANG; Ye-wei XIAO; Zhi-qiang FENG; Qiang-wen PAN

doi:10.13422/j.cnki.syfjx.20191303

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Effect and Mechanism of Celastrol on Endoplasmic Reticulum Stress in Hepatic L02 Cells of Nonalcoholic Fatty Liver Disease

Pharmacology | 更新时间：2021-02-09

- Effect and Mechanism of Celastrol on Endoplasmic Reticulum Stress in Hepatic L02 Cells of Nonalcoholic Fatty Liver Disease
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 14, Pages: 99-105(2019)
- 作者机构：
  
  西南医科大学，四川　泸州　 646000
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191303
  CLC： R2-0; R22; R285.5
- Published：20 July 2019，
  
  Published Online：19 March 2019，
  
  Received：05 November 2018，
- 稿件说明：
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Xia JIANG, Hui-ling JIANG, Fan YANG, et al. Effect and Mechanism of Celastrol on Endoplasmic Reticulum Stress in Hepatic L02 Cells of Nonalcoholic Fatty Liver Disease. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(14):99-105(2019)
DOI：

Xia JIANG, Hui-ling JIANG, Fan YANG, et al. Effect and Mechanism of Celastrol on Endoplasmic Reticulum Stress in Hepatic L02 Cells of Nonalcoholic Fatty Liver Disease. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(14):99-105(2019) DOI： 10.13422/j.cnki.syfjx.20191303.

摘要

目的：

通过雷公藤红素干预非酒精性脂肪肝L02细胞模型来探讨雷公藤红素调节肝L02细胞脂质代谢紊乱及非酒精性脂肪肝L02细胞内质网应激的相关机制。

方法：

将肝L02细胞分为空白组，模型组，雷公藤红素低剂量组(0.5 mg·L

－1

)，雷公藤红素高剂量组(1 mg·L

－1

)和辛伐他汀组(SIM，6 mg·L

－1

)进行培养。检测肝L02细胞内胆固醇(TC)和甘油三酯(TG)含量变化；用油红O染色观察肝L02细胞脂质沉积情况；逆转录聚合酶链式反应(RT-PCR)和蛋白免疫印迹法(Western blot)分别检测各组肝L02细胞中内质网应激(ERS)相关信号分子活化转录因子6(ATF6)，葡萄糖调节蛋白78(GRP78)，肌醇需求酶1(IRE1)，固醇调节元件结合蛋白裂解激活蛋白(SCAP)，固醇调节元件结合蛋白1c(SREBP-1c)和固醇调节元件结合蛋白-2(SREBP-2)的 mRNA转录以及蛋白表达水平。

结果：

非酒精性脂肪肝病(NAFLD)组肝L02细胞中TC及TG含量均高于空白组(

0.05)，雷公藤红素低、高剂量组及SIM 6 mg·L

－1

组肝L02细胞中TC及TG含量较NAFLD组均有不同程度减少(

0.05)。油红O染色显示NAFLD组肝L02细胞内含有大量红染脂质颗粒沉积，而雷公藤红素低、高剂量组及SIM 6 mg·L

－1

组红染脂质颗粒较NAFLD组均有不同程度减少。RT-PCR和Western blot结果显示，NAFLD组肝L02细胞内ERS相关信号分子ATF6，GRP78，IRE1，SCAP，SREBP-1c和SREBP-2的mRNA转录和蛋白表达水平均高于空白组(

0.05)；雷公藤红素低、高剂量组和SIM 6 mg·L

－1

组肝L02细胞内ATF6，GRP78，IRE1，SCAP，SREBP-1c和SREBP-2的mRNA转录及蛋白表达水平均低于NAFLD组(

0.05)。而雷公藤红素高剂量组与SIM 6 mg·L

－1

组间肝L02细胞内ATF6，GRP78，IRE1，SCAP，SREBP-1c和SREBP-2 mRNA转录及蛋白表达水平差异不具有统计学意义。

结论：

雷公藤红素可通过下调肝L02细胞ERS时相关信号分子ATF6，GRP78，IRE1，SCAP，SREBP-1c和SREBP-2的表达来减轻肝L02细胞脂质代谢紊乱，从而改善NAFLD。

Abstract

Objective:

To explore the effect of celastrol in inhibiting the lipid metabolism disorder in hepatic L02 cells and its possible mechanism on endoplasmic reticulum stress (ERS) of non-alcoholic fatty liver cells by intervening non-alcoholic fatty liver disease(NAFLD) cell model with celastrol.

Method:

Hepatic L02 cells were divided into control group

model group

low-dose celastrol treatment group (Cel 0.5 mg·L

-1

)

high-dose celastrol treatment group (Cel 1 mg·L

-1

) and simvastatin group (SIM 6 mg·L

-1

) for cultivation. The contents of total cholesterol (TC) and total triglyceride (TG) in hepatic L02 cells were detected

and the oil red staining was used to detected the lipid accumulation in hepatic L02 cells. Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to detect the mRNA and protein expression levels of endoplasmic reticulum stress (ERS)-related signal molecules activating transcription factor 6 (ATF6)

glucose regulated protein 78 (GRP78)

inositol-requiring enzyme 1 (IRE1)

sterol regulatory element-binding protein cleavage-activating protein (SCAP)

sterol regulatory element-binding protein-1c (SREBP-1c) and sterol regulatory element-binding protein-2 (SREBP-2) in hepatic L02 cell model respectively.

Result:

The contents of TC and TG in hepatic L02 cells of NAFLD group were significantly higher than those in control group (

0.05). The TC and TG contents in hepatic L02 cells of Cel 0.5 mg·L

-1

group

Cel 1 mg·L

-1

group and SIM 6 mg·L

-1

group were significantly lower than those in NAFLD group (

0.05). Oil red O staining showed that a large amount of red-stained lipid particles were deposited in the hepatic L02 cells of the NAFLD group

while the red-stained lipid particles in the Cel 0.5 mg·L

-1

group

the Cel 1 mg·L

-1

group

and the SIM 6 mg·L

-1

group were lower than the NAFLD group to different degrees. According to the results of RT-PCR and Western blot

the mRNA transcription and protein expression levels of ERS-related signaling molecules ATF6

GRP78

IRE1

SCAP

SREBP-1c and SREBP-2 in hepatic L02 cells of NAFLD group were higher than those of control group (

0.05). The mRNA transcription and protein expression levels of ERS-related signaling molecules ATF6

GRP78

IRE1

SCAP

SREBP-1c and SREBP-2 in hepatic L02 cells of Cel 0.5 mg·L

-1

group

Cel 1 mg·L

-1

group and SIM 6 mg·L

-1

group were lower than those of NAFLD group (

0.05). There was no significant difference in the mRNA transcription and protein expression levels of ATF6

GRP78

IRE1

SCAP

SREBP-1c and SREBP-2 between the Cel 1 mg·L

-1

group and the SIM 6 mg·L

-1

group.

Conclusion:

Celastrol can reduce the lipid metabolism disorder in hepatic L02 cells by down-regulating the expressions of ERS-related signaling molecules ATF6

GRP78

IRE1

SCAP

SREBP-1c and SREBP-2 in hepatic L02 cells

so as to improve NAFLD.

关键词

雷公藤红素非酒精性脂肪肝内质网应激肝L02细胞

Keywords

celastrolnon-alcoholic fatty liver diseaseendoplasmic reticulum stresshepatic L02 cells

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