Artesunate improves DDC-induced cholestatic liver fibrosis through PI3K/Akt/mTOR signaling pathway in mice

Wan TANG; Nan ZHAO; Xiaoxun ZHANG; Jin CHAI

doi:10.16016/j.2097-0927.202303033

您当前的位置：

首页 >

文章列表页 >

Artesunate improves DDC-induced cholestatic liver fibrosis through PI3K/Akt/mTOR signaling pathway in mice

Basic Medicine | 更新时间：2026-01-09

- Artesunate improves DDC-induced cholestatic liver fibrosis through PI3K/Akt/mTOR signaling pathway in mice
- Journal of Army Medical University Vol. 45, Issue 10, Pages: 1001-1009(2023)
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)第一附属医院消化内科，胆汁淤积肝病中心和代谢性肝病中心
- 作者简介：
- 基金信息：
  
  the National Natural Science Foundation for Distinguished Young Scholars of China(81922012);the Outstanding Youth Foundation of Chongqing(cstc2021jcyj-jqX0005)
- DOI：10.16016/j.2097-0927.202303033
  CLC： R282.71;R285.5;R575.22
- Received：07 March 2023，
  
  Revised：2023-03-19，
  
  Published：30 May 2023
- 稿件说明：
移动端阅览
Wan TANG, Nan ZHAO, Xiaoxun ZHANG, et al. Artesunate improves DDC-induced cholestatic liver fibrosis through PI3K/Akt/mTOR signaling pathway in mice[J]. Journal of Army Medical University, 2023, 45(10): 1001-1009.
DOI：

Wan TANG, Nan ZHAO, Xiaoxun ZHANG, et al. Artesunate improves DDC-induced cholestatic liver fibrosis through PI3K/Akt/mTOR signaling pathway in mice[J]. Journal of Army Medical University, 2023, 45(10): 1001-1009. DOI： 10.16016/j.2097-0927.202303033.

摘要

目的

探究青蒿琥酯(artesunate

ART)对3

5-二乙氧羰基-1

4-二氢-2

6-三甲吡啶(3

5-diethoxycarbonyl-1

4-dihydro-2

6-collidine

DDC)诱导的胆汁淤积性肝纤维化的作用及分子机制。

方法

将24只8周龄雄性C57BL/6J小鼠按随机数字表法分为3组：对照组(

=6)、0.1% DDC组(

=9)和0.1% DDC+ART组(

=9)。喂养含0.1% DDC的饲料2周，构建胆汁淤积小鼠模型，同时给予0.1% DDC+ART组小鼠青蒿琥酯(30.0 mg/kg，1次/d)肌肉注射。检测小鼠血清中肝损伤相关指标的水平；进行HE染色和天狼星红染色，根据Scheuer系统进行肝组织病理学评分。采用实时荧光定量PCR检测小鼠肝脏内α-SMA、Col1a1、Col1a2和Tgfβ1等表达水平，同时检测小鼠肝组织内羟脯氨酸(hydroxyproline

HYP)含量。在细胞实验中进一步探索青蒿琥酯减轻肝纤维化的机制，使用TGF-β联合ART处理LX2细胞，采用Western blot检测PI3K/Akt/mTOR信号通路相关蛋白的表达。使用ART和通路抑制剂共同处理LX2细胞后，检测肝星状细胞(hepatic stellate cell

HSC)活化标志物α-SMA的表达。

结果

青蒿琥酯给药明显减轻DDC诱导的胆汁淤积性肝损伤和肝纤维化(

0.05)，显著降低了胆汁淤积小鼠血清中ALT、AST、ALP和TBA的表达水平(

0.05)。青蒿琥酯抑制小鼠肝脏内纤维化相关指标α-SMA、Col1a1、Col1a2和Tgfβ1 mRNA的表达，并减少肝组织内HYP含量(

0.05)。青蒿琥酯可以抑制TGF-β诱导的LX2细胞内PI3K/Akt/mTOR信号通路的激活(

0.05)。PI3K/Akt/mTOR信号通路抑制剂增强青蒿琥酯对TGF-β诱导的HSC活化的抑制作用(

0.05)。

结论

青蒿琥酯能够明显改善DDC诱导的胆汁淤积性肝纤维化，它可通过调控PI3K/Akt/mTOR信号通路抑制HSC的活化，从而发挥其抗纤维化的作用。

Abstract

Objective

To explore the effect and molecular mechanism of artesunate (ART) on cholestatic liver fibrosis induced by 3

5-diethoxycarbonyl-1

4-dihydro-2

6-collidine (DDC).

Methods

8-week-old male C57BL/6J mice(

=24) were randomly divided into 3 groups: the control group(

=6)

the 0.1% DDC group(

=9) and the 0.1% DDC+ART group(

=9). Mice have been fed with diet containing 0.1% DDC for 2 weeks to establish cholestatic mouse model

and ART was given intramuscular injection (30.0 mg/kg) once a day in the 0.1% DDC+ART group. The indicators of liver injury in serum of mice were detected. HE staining and Sirius red staining were performed

and histopathological scores were obtained according to Scheuer scoring system. RT-qPCR was used to detect the levels of α-SMA

Col1a1

Col1a2 and Tgfβ1 in mouse liver. Hydroxyproline (HYP) content was detected with HYP detection kit. LX2 cells were treated with TGF-β combined with ART

and the proteins related to PI3K/Akt/mTOR pathway were detected by Western blot assay. The expression of α-SMA

hepatic stellate cell (HSC) activation marker

was detected after ART and pathway inhibitors were used to treated with LX2 cells.

Results

ART significantly ameliorated DDC-induced cholestatic liver injury and liver fibrosis (

0.05). ART markedly decreased the levels of ALT

AST

ALP and TBA in serum of mice (

0.05). ART inhibited the mRNA expression of α-SMA

Col1a1

Col1a2 and Tgfβ1

and reduced the content of HYP in liver tissues of mice with cholestasis(

0.05). In addition

ART obviously inhibited TGF-β-induced activation of PI3K/Akt/mTOR pathway in LX2 cells (

0.05). PI3K/Akt/mTOR signaling pathway inhibitors enhanced the inhibitory effect of HSC activation induced by TGF-β(

0.05).

Conclusion

ART observably improves DDC-induced cholestatic liver fibrosis

and it can inhibit HSC activation by regulating PI3K/Akt/mTOR signaling pathway

thus exerting anti-fibrosis effect.

关键词

Keywords

references

FICKERT P, POLLHEIMER M J, BEUERS U, et al . Characterization of animal models for primary sclerosing cholangitis (PSC)[J ] . J Hepatol, 2014, 60(6): 1290-1303. DOI: 10.1016/j.jhep.2014.02.006.

WANG C, MA C, FU K, et al . Phillygenin attenuates carbon tetrachloride-induced liver fibrosis via modulating inflammation and gut microbiota[J ] . Front Pharmacol, 2021, 12: 756924. DOI: 10.3389/fphar.2021.756924..

秦雨萌, 陈胜华. 肝纤维化进程中细胞外基质对肝星状细胞的作用及机制研究进展[J]. 实用肝脏病杂志, 2017, 20(3): 381-384. DOI: 10.3969/j.issn.1672-5069.2017.03.037.

QIN Y M, CHEN S H. Influence of extracellular matrix on hepatic stellate cells during liver fibrosis process[J]. J Pract Hepatol, 2017, 20(3): 381-384. DOI: 10.3969/j.issn.1672-5069.2017.03.037.

GHONEM N S, ASSIS D N, BOYER J L. Fibrates and cholestasis[J]. Hepatology, 2015, 62(2): 635-643. DOI: 10.1002/hep.27744.

FRANGOGIANNIS N. Transforming growth factor-β in tissue fibrosis[J]. J Exp Med, 2020, 217(3): e20190103. DOI: 10.1084/jem.20190103.

HAMIDI A, SONG J, THAKUR N, et al . TGF-β promotes PI3K-AKT signaling and prostate cancer cell migration through the TRAF6-mediated ubiquitylation of p85α[J ] . Sci Signal, 2017, 10(486): eaal4186[pii ] . DOI: 10.1126/scisignal.aal4186.

JAFARI M, GHADAMI E, DADKHAH T, et al . PI3k/AKT signaling pathway: erythropoiesis and beyond[J ] . J Cell Physiol, 2019, 234(3): 2373-2385. DOI: 1 0.1002/jcp.27262.

SHIGEMATSU T, TAJIMA S, FU R, et al . The mTOR inhibitor everolimus attenuates tacrolimus-induced renal interstitial fibrosis in rats[J ] . Life Sci, 2022, 288: 120150. DOI: 10.1016/j.lfs.2021.120150.

陈海亮, 刘毅, 邹中元, 等. 青蒿琥酯与奎宁治疗疟疾的疗效比较[J]. 江西医药, 2014, 49(1): 57-59. DOI: 10.3969/j.issn.1006-2238.2014.01.026.

CHEN H L, LIU Y, ZOU Z Y, et al . Comparison between artesunate and quinine in the treatment of malaria[J ] . Jiangxi Med J, 2014, 49(1): 57-59. DOI: 10.3969/j.issn.1006-2238.2014.01.026.

蒋沅岐, 董玉洁, 周福军, 等. 青蒿素及其衍生物的研究进展[J]. 中草药, 2022, 53(2): 599-608. DOI: 10.7501/j.issn.0253-2670.2022.02.030.

JIANG Y Q, DONG Y J, ZHOU F J, et al . Research progress on artemisinin and its derivatives[J ] . Chin Tradit Herb Drugs, 2022, 53(2): 599-608. DOI: 10.7501/j.issn.0253-2670.2022.02.030.

KONG Z Y, LIU R, CHENG Y. Artesunate alleviates liver fibrosis by regulating ferroptosis signaling pathway[J]. Biomed Pharmacother, 2019, 109: 2043-2053. DOI: 10.1016/j.biopha.2018.11.030.

郭辉, 肖刚, 孟昕珂, 等. 基于TGF-β1/Smads信号通路研究胆通颗粒对胆汁淤积性肝纤维化的影响[J]. 现代中西医结合杂志, 2020, 29(18): 1944-1947, 2024. DOI: 10.3969/j.issn.1008-8849.2020.18.002.

GUO H, XIAO G, MENG X K, et al . Study on the effect of Dantong Granule on cholestatic liver fibrosis based on TGF-β1/Smads signal pathway[J ] . Mod J Integr Tradit Chin West Med, 2020, 29(18): 1944-1947, 2024. DOI: 10.3969/j.issn.1008-8849.2020.18.002.

HIGASHI T, FRIEDMAN S L, HOSHIDA Y. Hepatic stellate cells as key target in liver fibrosis[J]. Adv Drug Deliv Rev, 2017, 121: S0169-S409X(17)30063-7[pii]. DOI: 10.1016/j.addr.2017.05.007.

YANG X L, ZHENG Y D, LIU L, et al . Progress on the study of the anticancer effects of artesunate[J ] . Oncol Lett, 2021, 22(5): 750. DOI: 10.3892/ol.2021.13011.

SINCLAIR D, DONEGAN S, ISBA R, et al . Artesunate versus quinine for treating severe malaria[J ] . Cochrane Database Syst Rev, 2012, 2012(6): CD005967. DOI: 10.1002/14651858.CD005967.pub4.

LIU Y J, HUANG G J, MO B W, et al . Artesunate ameliorates lung fibrosis via inhibiting the Notch signaling pathway[J ] . Exp Ther Med, 2017, 14(1): 561-566. DOI: 10.3892/etm.2017.4573..

SHUANG, SHEN. Artesunate alleviates schistosomiasis-induced liver fibrosis by downregulation of mitochondrial complex Ⅰ subunit NDUFB8 and complex Ⅲ subunit UQCRC2 in hepatic stellate cells[J]. Acta Trop, 2021, 214: 105781. DOI: 10.1016/j.actatropica.2020.105781.

CAO J, WANG W, LI Y, et al . Artesunate attenuates unilateral ureteral obstruction-induced renal fibrosis by regulating theexpressions of bone morphogenetic protein-7 and uterine sensitization-associated gene-1 in rats[J ] . Int Urol Nephrol, 2016, 48(4): 619-629. DOI: 10.1007/s11255-016-1232-0.

张波, 谈仁秀, 黄芝月, 等. MMI-0100对DDC诱导的胆汁淤积性肝损伤的治疗作用研究[J]. 中国现代应用药学, 2022, 39(13): 1692-1697. DOI: 10.13748/j.cnki.issn1007-7693.2022.13.005.

ZHANG B, TAN R X, HUANG Z Y, et al . Study on therapeutic effect of MMI-0100 against DDC-induced cholestatic liver injury[J ] . Chin J Mod Appl Pharm, 2022, 39(13): 1692-1697. DOI: 10.13748/j.cnki.issn1007-7693.2022.13.005.

李欣泽, 王彦君, 李杨, 等. 转化生长因子-β信号通路在疾病发生发展中作用的研究状况[J]. 中国临床药理学杂志, 2022, 38(16): 1958-1962. DOI: 10.13699/j.cnki.1001-6821.2022.16.026.

LI X Z, WANG Y J, LI Y, et al . Study on the role of transforming growth factor-β/Smads signaling pathway in the occur rence and development of diseases[J ] . Chin J Clin Pharmacol, 2022, 38(16): 1958-1962. DOI: 10.13699/j.cnki.1001-6821.2022.16.026.

LEI Y, WANG Q L, SHEN L, et al . MicroRNA-101 suppresses liver fibrosis by down-regulating PI3K/Akt/mTOR signaling pathway[J ] . Clin Res Hepatol Gastroenterol, 2019, 43(5): 575-584. DOI: 10.1016/j.clinre.2019.02.003.

JI D, ZHAO Q, QIN Y W, et al . Germacrone improves liver fibrosis by regulating the PI3K/AKT/mTOR signalling pathway[J ] . Cell Biol Int, 2021, 45(9): 1866-1875. DOI: 10.1002/cbin.11607.

XU L, HUI A Y, ALBANIS E, et al . Human hepatic stellate cell lines, LX-1 and LX-2: new tools for analysis of hepatic fibrosis[J ] . Gut, 2005, 54(1): 142-151. DOI: 10.1136/gut.2004.042127.

MEJIAS M, GALLEGO J, NARANJO-SUAREZ S, et al . CPEB4 increases expression of PFKFB3 to induce glycolysis and activate mouse and human hepatic stellate cells, promoting liver fibrosis[J ] . Gastroenterology, 2020, 159(1): 273-288. DOI: 10.1053/j.gastro.2020.03.008.

LUO X J, LI H G, MA L Q, et al . Expression of STING is increased in li ver tissues from patients with NAFLD and promotes macrophage-mediated hepatic inflammation and fibrosis in mice[J ] . Gastroenterology, 2018, 155(6): 1971-1984. e4. DOI: 10.1053/j.gastro.2018.09.010.

GAUL S, LESZCZYNSKA A, ALEGRE F, et al . Hepatocyte pyroptosis and release of inflammasome particles induce stellate cell activation and liver fibrosis[J ] . J Hepatol, 2021, 74(1): 156-167. DOI: 10.1016/j.jhep.2020.07.041.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

High-salt diet promotes the progression of fibrosis in metabolic dysfunction-associated steatohepatitis via activating the PI3K/Akt/mTOR signaling pathway

Artesunate reduces IL-6 production in primary mouse hepatocytes induced by conjugated bile acids through inhibiting p-p65 signaling pathway

CX3CR1 silencing-mediated macrophage polarization attenuates CCl₄ -induced liver fibrosis in mice

Melatonin inhibits bile acids-initiated Cxcl2 expression through JNK/c-Jun signalling pathway

Effect of S100A6 on activation of hepatic stellate cells in mice

Related Author

TANG Yin

ZHANG Jie

MAO Cui

XIONG Tao

XIE Chuanming

ZHANG Leida

Min LIAO

Xuan LI

Related Institution

Department of Hepatobiliary Surgery， First Affiliated Hospital（Southwest Hospital）， Army Medical University （Third Military Medical University）

Center for Cholestatic Liver Diseases, Department of Gastroenterology, First Affiliated Hospital, Army Medical University (Third Military Medical University)

Department of Gastroenterology, Tongren Hospital Affiliated to Wuhan University (Wuhan Third Hospital)

Center for Metabolic Liver Diseases and Center for Cholestatic Liver Diseases, Department of Gastroenterology, First Affiliated Hospital, Army Medical University (Third Military Medical University)

Department of Laboratory Medicine, the First Affiliated Hospital of Chongqing Medical University

⁰