Effect of Shenling Baizhusan on Alcoholic Liver Injury in Rats by Regulating Ferroptosis Based on Nrf2 Signaling Pathway

ZHOU Xiangyu; ZHOU Sufang; LI Yuru; CAI Shiqin; LIAO Jiajia; YE Zuoyu

doi:10.13422/j.cnki.syfjx.20221907

您当前的位置：

首页 >

文章列表页 >

Effect of Shenling Baizhusan on Alcoholic Liver Injury in Rats by Regulating Ferroptosis Based on Nrf2 Signaling Pathway

更新时间：2023-02-07

- Effect of Shenling Baizhusan on Alcoholic Liver Injury in Rats by Regulating Ferroptosis Based on Nrf2 Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 5, Pages: 104-113(2023)
- 作者机构：
  
  1.贵州中医药大学第一临床医学院，贵阳 550002
  2.贵州中医药大学第一附属医院，贵阳 550001
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20221907
  CLC： R2-0;R22;R285.5;R289;R33
- Published：05 March 2023，
  
  Published Online：19 October 2022，
  
  Received：31 July 2022，
- 稿件说明：
扫描看全文
周相宇,周素芳,李玉茹等.基于Nrf2信号通路探究参苓白术散调节铁死亡对酒精性肝损伤大鼠的影响[J].中国实验方剂学杂志,2023,29(05):104-113.

ZHOU Xiangyu,ZHOU Sufang,LI Yuru,et al.Effect of Shenling Baizhusan on Alcoholic Liver Injury in Rats by Regulating Ferroptosis Based on Nrf2 Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(05):104-113.
周相宇,周素芳,李玉茹等.基于Nrf2信号通路探究参苓白术散调节铁死亡对酒精性肝损伤大鼠的影响[J].中国实验方剂学杂志,2023,29(05):104-113. DOI： 10.13422/j.cnki.syfjx.20221907.

ZHOU Xiangyu,ZHOU Sufang,LI Yuru,et al.Effect of Shenling Baizhusan on Alcoholic Liver Injury in Rats by Regulating Ferroptosis Based on Nrf2 Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(05):104-113. DOI： 10.13422/j.cnki.syfjx.20221907.

摘要

目的

观察参苓白术散通过调节核因子E

相关因子2（Nrf2）信号通路的干预调节铁死亡对酒精性肝损伤大鼠的影响。

方法

将40只SD大鼠随机分为模型组、多烯磷脂酰胆碱组、参苓白术散高、中、低剂量组，每组8只，另取8只SD大鼠作为正常组。模型组、多烯磷脂酰胆碱组、参苓白术散高、中、低剂量组予以10 mL·kg

-1

灌胃造模，正常组予以等体积蒸馏水灌胃；每日灌酒4 h后，予以多烯磷脂酰胆碱组143.64 mg·kg

-1

药物灌胃、参苓白术散高、中、低剂量组分别予以15、7.5、3.75 mg·kg

-1

的药物，正常组与模型组予以等体积蒸馏水灌胃，灌胃连续6周。全自动生化仪检测丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、谷氨酰转肽酶（GGT）、总胆固醇（TC）、甘油三酯（TG）水平，酶联免疫吸附测定法（ELISA）检测肿瘤坏死因子-

（TNF-

）、白细胞介素-1

（IL-1

）水平，生化检测法检测脂多糖（LPS）、丙二醇（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）、铁离子（Fe

）水平，苏木素-伊红（HE）和油红O染色观察肝脏病理变化，实时荧光定量聚合酶链式反应（Real-time PCR）检测Nrf2、血红素加氧酶-1（HO-1）、谷胱甘肽过氧化物酶4（GPX4）、铁蛋白重链1（FTH1）、核转录因子-

B（NF-

B） mRNA表达，蛋白免疫印迹法（Western blot）检测Nrf2、HO-1、GPX4、FTH1、p65、磷酸化（p）-p65蛋白表达。

结果

与正常组比较，模型组肝功能（ALT、AST、GGT）、血脂（TC、TG）水平明显升高（

0.05），肝脏出现明显脂肪变性，有大量脂肪沉积，氧化应激水平及炎症因子明显升高（

0.05），Fe

水平明显增高（

0.05），Nrf2、HO-1、GPX4、FTH1蛋白表达明显下调（

0.05），p65、p-p65蛋白表达明显上调（

0.05），Nrf2、HO-1、GPX4、FTH1 mRNA表达明显下调（

0.05），NF-

B mRNA表达明显上调（

0.05）；与模型组比较，参苓白术散高、中剂量组肝功能（ALT、AST、GGT）、血脂（TC、TG）水平明显下调（

0.05），肝脏脂肪变性明显改善，脂肪沉积明显减少，氧化应激水平以及炎症因子明显降低（

0.05），Fe

水平明显降低（

0.05），Nrf2、HO-1、GPX4、FTH1蛋白表达明显上调（

0.05），p65、p-p65蛋白表达明显下调（

0.05），Nrf2、HO-1、GPX4、FTH1 mRNA表达明显上调（

0.05）、NF-

B mRNA表达明显下调（

0.05）。

结论

参苓白术散能够有效减少ALD大鼠肝损伤，调节脂肪变性与脂肪沉积，并在肝脏内发挥抗氧化与抗炎作用；其作用机制可能与上调Nrf2信号通路改善氧化应激的途径抑制肝细胞内铁死亡发生有关。

Abstract

Objective

To observe the effect of Shenling Baizhusan on the intervention of the nuclear factor erythroid 2-related factor 2 （Nrf2） signaling pathway by regulating ferroptosis in rats with alcoholic liver injury.

Method

Forty SD rats were randomly divided into model group， polyene phosphatidylcholine group， and high， medium， and low-dose Shenling Baizhusan groups， with 8 rats in each group. Another 8 SD rats were taken as blank group. The model group， polyene phosphatidylcholine group， high， medium， and low-dose Shenling Baizhusan groups were given 10 mL·kg

-1

liquor by gavage for modeling， and the blank group was given equal volume of distilled water by gavage. After 4 h of daily alcoholic administration， 143.64 mg·kg

-1

of polyene phosphatidylcholine group was given to the polyene phosphatidylcholine group， 15， 7.5， 3.75 mg·kg

-1

of Shenling Baizhusan were given to Shenling Baizhusan high， medium， and low-dose groups， respectively， and the blank group and the model group were given equal volume of distilled water. The gavage lasted for 6 weeks. The levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， glutamyl transpeptidase （GGT）， total cholesterol （TC）， and triglyceride （TG） were detected by automatic biochemical analyzer. The levels of tumor necrosis factor-

（TNF-

） and interleukin-

（IL-

） were detected by the enzyme-linked immunosorbent assay （ELISA）. The levels of lipopolysaccharide （LPS）， malondialdehyde （MDA）， superoxide dismutase （SOD）， glutathione （GSH）， and Fe

were detected by biochemical assay. The pathological changes in the liver were observed by hematoxylin-eosin （HE） staining and oil red O staining. The mRNA expression levels of Nrf2， heme oxygenase-1 （HO-1）， glutathione peroxidase 4 （GPX4）， ferritin heavy polypeptide 1 （FTH1）， and nuclear factor-

B （NF-

B） were detected by Real-time polymerase chain reaction （Real-time PCR）. The protein expression levels of Nrf2， HO-1， GPX4， FTH1， p65， and phosphorylation （p）-p65 were detected by Western blot.

Result

As compared with the blank group， the levels of liver function （ALT， AST， and GGT） and blood lipids （TC and TG） in the model group were significantly increased （

0.05）. The liver showed obvious steatosis， with a large number of fat deposition， the oxidative stress and inflammatory factors were significantly increased （

0.05）， and the level of Fe

was significantly increased in model group （

0.05）. The protein expression levels of Nrf2， HO-1， GPX4， and FTH1 was significantly down-regulated （

0.05）， and those of p65 and p-p65 was significantly up-regulated in the model group （

0.05）. The mRNA expression levels of Nrf2， HO-1， GPX4， and FTH1 were significantly down-regulated （

0.05）， and the mRNA expression level of NF-

B was significantly up-regulated （

0.05）. As compared with the model group， the levels of liver function （ALT， AST， and GGT） and blood lipids （TC and TG） in the high-dose and medium-dose Shenling Baizhusan groups were significantly decreased （

0.05）， liver steatosis was significantly improved， fat deposition was significantly reduced， oxidative stress and inflammatory factors were significantly decreased （

0.05 ）， and Fe

level was significantly decreased （

0.05）. In the high-dose and medium-dose Shenling Baizhusan， the protein expression levels of Nrf2， HO-1， GPX4， and FTH1 were significantly up-regulated （

0.05）， and those of p65， p-p65 were significantly down-regulated （

0.05）. The mRNA expression levels of Nrf2， HO-1， GPX4， and FTH1 were significantly up-regulated （

0.05）， and the mRNA expression level of NF-

B was significantly down-regulated （

0.05）.

Conclusion

Shenling Baizhusan can effectively reduce liver injury in rats with ALD， regulate steatosis and fat deposition， and play an antioxidant and anti-inflammatory role in the liver. Its mechanism may be related to the inhibition of ferroptosis in hepatocytes by up-regulating the Nrf2 signaling pathway to improve oxidative stress

关键词

参苓白术散酒精性肝损伤核因子E2相关因子2（Nrf2）氧化应激铁死亡

Keywords

Shenling Baizhusanalcoholic liver injurynuclear factor erythroid 2-related factor 2 （Nrf2）oxidative stressferroptosis

references

中华医学会肝病学分会脂肪肝和酒精性肝病学组，中国医师协会脂肪性肝病专家委员会.酒精性肝病防治指南（2018更新版）［J］.中华肝脏病杂志，2018，26（3）：188-194.

BYASS P.The global burden of liver disease： A challenge for methods and for public health［J］.BMC Med，2014，12：159.

XIAO J，WANG F，WONG N K，et al.Global liver disease burdens and research trends： Analysis from a Chinese perspective［J］.J Hepatol，2019，71（1）：212-221.

MUELLER S，PECCERELLA T，QIN H，et al.Carcinogenic etheno DNA adducts in alcoholic liver disease： Correlation with cytochrome P-4502E1 and fibrosis［J］.Alcohol Clin Exp Res，2018，42（2）：252-259.

LV X，CHEN Z，LI J，et al.Caffeine protects against alcoholic liver injury by attenuating inflammatory response and oxidative stress［J］.Inflamm Res，2010，59（8）：635-645.

PETRASEK J，BALA S，CSAK T，et al.IL-1 receptor antagonist ameliorates inflammasome-dependent alcoholic steatohepatitis in mice［J］.J Clin Invest，2012，122（10）：3476-3489.

DIXON S J，LEMBERG K M，LAMPRECHT M R，et al.Ferroptosis： An iron-dependent form of nonapoptotic cell death［J］.Cell，2012，149（5）：1060-1072.

JIANG X，STOCKWELL B R，CONRAD M.Ferroptosis： Mechanisms， biology and role in disease［J］.Nat Rev Mol Cell Biol，2021，22（4）：266-282.

HARRISON-FINDIK D D，SCHAFER D，KLEIN E，et al.Alcohol metabolism-mediated oxidative stress down-regulates hepcidin transcription and leads to increased duodenal iron transporter expression［J］.J Biol Chem，2006，281（32）：22974-22982.

卢殿强，邵铭.邵铭教授从脾论治酒精性肝病［J］.世界中西医结合杂志，2013，8（11）：1160-1161.

周素芳，刘闯，李艺锋，等.参苓白术颗粒对酒精性肝病大鼠肝肠损伤的改善作用［J］.中华中医药学刊，2021，39（7）：4-7，后插1和2.

王峰.加味参苓白术散对非酒精性脂肪肝疗效分析［J］.中国中医基础医学杂志，2013，19（12）：1489，1504.

吕锦珍，徐拥建，胡世平，等.参苓白术散对NAFLD大鼠肝细胞mTORC1/STAT3信号通路的影响［J］.中国实验方剂学杂志，2020，26（2）：6-12.

卢广英，邢训颜，王嘉昀，等.经典名方参苓白术散的研究进展及质量标志物的预测分析［J］.中国中药杂志，2022，47（19）：5171-5181.

赖靖江，禚孝丽，蒋凤仙，等.参苓白术散在多种疾病模型中的潜在作用机制［J］.中国实验方剂学杂志，2022，28（20）：267-273.

中华中医药学会中药实验药理专业委员会.酒精性肝损伤动物模型制备规范（草案）［J］.中华中医药杂志，2018，33（3）：1000-1003.

张茹，曲中原，杜娟.基于Nrf2-HO-1/CYP2E1通路探讨五指毛桃醇提取物对酒精性肝损伤小鼠的抗氧化保护机制［J］.中药新药与临床药理，2021，32（12）：1769-1775.

彭景华，方志红，崔剑巍，等.健脾活血方对酒精复合内毒素脂多糖诱导的肝损伤大鼠库普弗细胞活化信号通路的干预［J］.中西医结合学报，2007，5（3）：302-306.

方俐晖，郭志玲，张轶斐，等.田德禄教授三期三脏辨治酒精性肝病经验［J］.现代中医临床，2021，28（5）：38-42.

刘江凯.赵文霞分期辨治酒精性肝病经验介绍［J］.新中医，2021，53（18）：201-204.

朱平生，焦炎杰，付双楠，等.酒精致大鼠肝损伤早期血清生物标志物水平的变化规律［J］.中国实验方剂学杂志，2019，25（2）：129-133.

PARKER R，KIM S J，GAO B.Alcohol， adipose tissue and liver disease： Mechanistic links and clinical considerations［J］.Nat Rev Gastroenterol Hepatol，2018，15（1）：50-59.

ZHONG W，ZHAO Y，TANG Y，et al.Chronic alcohol exposure stimulates adipose tissue lipolysis in mice： Role of reverse triglyceride transport in the pathogenesis of alcoholic steatosis［J］.Am J Pathol，2012，180（3）：998-1007.

DREGER H，WESTPHAL K，WILCK N，et al.Protection of vascular cells from oxidative stress by proteasome inhibition depends on Nrf2［J］.Cardiovasc Res，2009，85：395-403.

HU X，ZHOU R，LI H，et al.Alterations of gut microbiome and serum metabolome in coronary artery disease patients complicated with non-alcoholic fatty liver disease are associated with adverse cardiovascular outcomes［J］.Front Cardiovasc Med，2021，8：805812.

DINARELLO C A.Immunological and inflammatory functions of the interleukin-1 family［J］.Annu Rev Immunol，2009，27：519-550.

PETRASEK J，BALA S，CSAK T，et al.IL-1 receptor antagonist ameliorates inflammasome-dependent alcoholic steatohepatitis in mice［J］.J Clin Invest，2012，122（10）：3476-3489.

KANY S，JANICOVA A，RELJA B.Innate immunity and alcohol［J］.J Clin Med，2019，8（11）：234.

KIM A，BELLAR A，MCMULLEN M R，et al.Functionally diverse inflammatory responses in peripheral and liver monocytes in alcohol-associated hepatitis［J］.Hepatol Commun，2020，4（10）：1459-1476.

XU M J，ZHOU Z，PARKER R，et al.Targeting inflammation for the treatment of alcoholic liver disease［J］.Pharmacol Ther，2017，180：77-89.

LI X L，JIANG W，FAN W M，et al.Role of gut microbiota in the treatment of nonalcoholic fatty liver disease with traditional Chinese medicine［J］.Acta Pharm Sin（药学学报），2020，55：15-24.

MOHS A，OTTO T，SCHNEIDER K M，et al.Hepatocyte-specific Nrf2 activation controls fibrogenesis and carcinogenesis in steatohepatitis［J］.J Hepatol，2021，74（3）：638-648.

LI L，FU J，SUN J，et al.Is Nrf2-ARE a potential target in NAFLD mitigation？［J］.Curr Opin Toxicol，2019，13：35-44.

SCHÖDEL J，RATCLIFFE P J.Mechanisms of hypoxia signalling： New implications for nephrology［J］.Nat Rev Nephrol，2019，15：641-659.

SILVA-GOMES S，SANTOS A G，CALDAS C，et al.Transcription factor Nrf2 protects mice against dietary iron-induced liver injury by preventing hepatocytic cell death［J］.J Hepatol，2014，60（2）：354-361.

RISHI G，SUBRAMANIAM V N.The liver in regulation of iron homeostasis［J］.Am J Physiol Gastrointest Liver Physiol，2017，313：G157-G165.

HARRIS I S，DENICOLA G M.The complex interplay between antioxidants and ROS in cancer［J］.Trends Cell Biol，2020，30，440-451.

RAN Q，MOZOLEWSKA P.Gpx4 and Ferroptosis.In ferroptosis in health and disease［M］.Cham：Springer， 2019：99-109.

HOU W，XIE Y，SONG X，et al.Autophagy promotes ferroptosis by degradation of ferritin［J］.Autophagy，2016，12（8）：1425-1428.

LIU C Y，WANG M，YU H M，et al.Ferroptosis is involved in alcohol-induced cell death in vivo and in vitro［J］.Biosci Biotechnol Biochem，2020，84（8）：1621-1628.

LIU J，HE H，WANG J，et al.Oxidative stress-dependent frataxin inhibition mediated alcoholic hepatocytotoxicity through ferroptosis［J］.Toxicology，2020，445：152584.

岳亚光，檀薇薇，王文川，等.二甲双胍经Nrf2-Gpx4通路抑制铁死亡并减轻非酒精性脂肪性肝病大鼠的肝损伤［J］.中国病理生理杂志，2021，37（10）：1848-1857.

张新，陈文娜，宋囡，等.丹蒌片通过铁死亡途径减轻非酒精性脂肪性肝病模型小鼠肝脏氧化损伤［J］.中国病理生理杂志，2021，37（12）：2180-2188.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Experimental Study on Regulation of Nrf2/HO-1 by Linalool to Inhibit Hepatic Injury Induced by Aflatoxin B₁

Exploring Detoxication Mechanism of Dioscoreae Bulbiferae Rhizoma Processed with Phaseoli Radiati Semen Juice Based on Target Organ Ferroptosis

Earthworm Protein Protects Vascular Endothelial Function in Spontaneously Hypertensive Rats via PI3K/Akt/Nrf2 Signaling Pathway

Effect of Buyang Huanwutang on Ferroptosis in Diabetic Kidney Disease Mice

Effect of Glycyrrhizae Radix et Rhizoma-containing Serum on LPS-induced Inflammation in Caco2 Cells Based on Inhibition of Ferroptosis by Nrf2/HO-1 Pathway

Related Author

WANG Meng

XUE Chunmiao

HUANG Xin

LIU Wenhui

GAO Ruoyu

BAI Xuehui

HUA Guodong

ZHU Baochen

Related Institution

Dongzhimen Hospital，Beijing University of Chinese Medicine

School of Chinese Materia Medica， Beijing University of Chinese Medicine

Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Province and Ministry of Education，Collaborative Innovation Center of Research and Development on the Whole Industry Chain of Yu-Yao，Henan Province， Henan University of Chinese Medicine

The First Affiliated Hospital of Henan University of Chinese Medicine

Gansu University of Chinese Medicine

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