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

您当前的位置：

首页 >

文章列表页 >

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
- Received：05 November 2018，
  
  Published Online：19 March 2019，
  
  Published：20 July 2019
- 稿件说明：
移动端阅览
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, 2019, 25(14): 99-105.
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, 2019, 25(14): 99-105. 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

Keywords

references

Pettinelli P , Obregón A M , Videla L A . Molecular mechanisms of steatosis in nonalcoholic fatty liver disease [J]. Nutr Hosp , 2011 , 26 ( 3 ): 441 - 450 .

王健，姜曼，辛永宁，等．内质网应激在非酒精性脂肪性肝病发病机制中的作用 [J]．临床肝胆病杂志， 2014 ， 30 ( 11 )： 1229 - 1232 ．

刘佳，李传飞，宁波，等．内质网应激通过SCAP/SREBP-1c调控肝L02肝L02细胞脂质合成代谢 [J]．第三军医大学学报， 2015 ， 37 ( 5 )： 443 - 448

FANG D , WAN Y , SHEN W , et al . Endoplasmic reticulum stress leads to lipid accumulation through upregulation of SREBP-1c in normal hepatic and hepatoma cells [J]. Mol Cell Biochem , 2013 , 381 ( 1/2 ): 127 - 137 .

Ozcan U , CAO Q , Yilmaz E , et al . Endoplasmic reticulum stress links obesity, insulin action, and type2 diabetes [J]. Science , 2004 , 306 ( 5695 ): 457 - 461 .

ZHOU X , HAN D , YANG X , et al . Glucose regulated protein 78 is potentially an important player in the development of nonalcoholic steatohepatitis [J]. Gene , 2017 , 637 : 138 - 144 .

REN L P , YU X , SONG G Y , et al . Impact of activating transcription factor 4 signaling on lipogenesis in HepG2 cells [J]. Mol Med Rep , 2016 , 14 ( 2 ): 1649 - 1658 .

LI X , XU Z , WANG S , et al . Emodin ameliorates hepatic steatosis through endoplasmic reticulum stress-sterol regulatory element binding protein 1c pathway in liquid-fructose feeding rats [J]. Hepatol Res , 2016 , 46 ( 3 ): E105 - E117 .

徐佳，伍春莲，黄杰．雷公藤红素通过影响Akt信号通路和整合素表达抑制肺癌细胞的转移 [J]．中国中药杂志， 2015 ， 40 ( 6 )： 1129 - 1133 ．

王海波，倪腾洋，冯俊，等．南蛇藤提取物通过Cofilin1抑制胃癌BGC-823细胞转移的机制 [J]．中国实验方剂学杂志， 2018 ， 24 ( 19 )： 112 - 116 ．

钟点，陈渊，赵伟．雷公藤红素抗炎及免疫抑制的研究进展 [J]．药物生物技术， 2018 ， 25 ( 1 )： 64 - 69 ．

陶玲，管咏梅，陈丽华，等．雷公藤配伍减毒研究进展 [J]．中国实验方剂学杂志， 2018 ， 24 ( 4 )： 229 - 234 ．

Venkatesha S H , Moudgil K D . Celastrol and its role in controlling chronic diseases [J]. Adv Exp Med Biol , 2016 , 928 : 267 - 289 .

LIU J , Lee J , Salazarhernandez M A , et al . Treatment of obesity with celastrol [J]. Cell , 2015 , 161 ( 5 ): 999 - 1011 .

王万东，陈东风．非酒精性脂肪变性肝细胞模型中ATF4基因的表达及其意义 [J]．第三军医大学学报， 2010 ， 32 ( 14 )： 7 - 10 ．

YU X , TAO W , JIANG F , et al . Celastrol attenuates hypertension-induced inflammation and oxidative stress in vascular smooth muscle cells via induction of heme oxygenase-1 [J]. Am J Hypertens , 2010 , 23 ( 8 ): 895 - 903 .

Kim J E , MI H L , Nam D H , et al . Celastrol, an NF-κB inhibitor, improves insulin resistance and attenuates renal injury in db/db mice [J]. PLoS One , 2013 , 8 ( 4 ): e62068 .

Rutkowski D T , WU J , Back S H , et al . UPR pathways combine to prevent hepatic steatosis caused by ER stress-mediated suppression of transcriptional master regulators [J]. Dev Cell , 2008 , 15 ( 6 ): 829 - 840 .

Goldstein J L , DeBose-Boyd R A , Brown M S . Protein sensors for membrane sterols [J]. Cell , 2006 , 124 ( 1 ): 35 - 46 .

Zorov D B , Juhaszova M , Sollott S J . Mitochondrial ROS-induced ROS release: an update and review [J]. Biochim Biophys Acta , 2006 , 1757 ( 5 ): 509 - 517 .

皮振钧，李斌，刘俊科，等．内质网应激与肝细胞脂类代谢 [J]．现代生物医学进展， 2013 ， 13 ( 23 )： 4579 - 4582 ．

ZHANG C , CHEN X , ZHU R M , et al . Endoplasmic reticulum stress is involved in hepatic SREBP-1c activation and lipid accumulation in fructose-fed mice [J]. Toxicol Lett , 2012 , 212 ( 3 ): 229 - 240 .

万颖．内质网应激通过SREBP-1c-FAS途径影响肝L02细胞脂质沉积 [D]．重庆：重庆医科大学， 2013 ．

Colgan S M , TANG D , Werstuck G H , et al . Endoplasmic reticulum stress causes the activation of sterol regulatory element binding protein-2 [J]. Int J Biochem Cell Biol , 2007 , 39 ( 10 ): 1843 - 1851 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Mechanism of Ethoxysanguinarine in Regulating IRE1/RIDD Signaling Pathway to Inhibit Endoplasmic Reticulum Stress and Alleviate Cardiomyocyte Apoptosis

Effect of Shenkang Injection on Podocyte Apoptosis and GRP78/CHOP Signaling Pathway in db/db Mice with Diabetic Kidney Disease Based on Endoplasmic Reticulum Stress

Buzhong Yiqitang Regulates Endoplasmic Reticulum Stress to Attenuate Cisplatin Resistance in Non-small Cell Lung Cancer via Nrf2/ROS Pathway

Buzhong Yiqitang Regulates Endoplasmic Reticulum Stress via Nrf2/ROS/PERK/CHOP Signaling Pathway to Attenuate Cisplatin Resistance in NSCLC

Mechanisms of Spermatogenic Dysfunction and Sperm Quality Degradation Induced by Propylthiouracil via Endoplasmic Reticulum Stress in Rats and Interventional Effect of Shugan Bushen Yulin Decoction

Related Author

SHANG Zucheng

LI Hongzheng

LI Mengfan

SUN Wen

LIN Guosheng

SHEN Aling

CAI Yanmo

WANG Sitong

Related Institution

Fujian University of Traditional Chinese Medicine（TCM）

Guang'anmen Hospital， China Academy of Chinese Medical Sciences

Academy of Integrative Medicine，Fujian University of TCM

School of Traditional Chinese Medicine， Beijing University of Chinese Medicine

Institute of Chinese Materia Medica，China Academy of Chinese Medical Sciences

AI问答

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰