Ethyl Acetate Fraction of <italic style="font-style: italic">Dicliptera chinensis</italic> (L.) <italic style="font-style: italic">Juss</italic>.Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathways

Yuan LIU; Yan-meng BI; Ting PAN; Ting ZENG; Chan MO; Bing SUN; Lei GAO; Zhi-ping LYU

doi:10.1007/s11655-021-3298-5

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Ethyl Acetate Fraction of Dicliptera chinensis (L.) Juss.Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathways

Original Article | Updated：2022-03-18

- Ethyl Acetate Fraction of Dicliptera chinensis (L.) Juss.Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathways
- Chinese Journal of Integrative Medicine Vol. 28, Issue 1, Pages: 60-68(2022)
- Affiliations：
  
  1.College of Integrated Traditional Chinese and Western Medicine, Jining Medical University, Jining ,Shandong Province(272000), China
  2.School of Traditional Chinese Medicine,Southern Medical University, Guangzhou (510515), China
  3.Department of Traditional Chinese Medicine, Affiliated Hospital of Jining Medical University, Jining ,Shandong Province (272000),China
- Author bio：
  
  Prof. LYU Zhi-ping, E-mail: lzp48241@126.com
- Funds：
- DOI：10.1007/s11655-021-3298-5
  CLC：
- Published：01 January 2022，
  
  Published Online：08 June 2021，
  
  Accepted：17 November 2020
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Yuan LIU, Yan-meng BI, Ting PAN, et al. Ethyl Acetate Fraction of Dicliptera chinensis (L.) Juss.Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathways. [J]. Chinese Journal of Integrative Medicine 28(1):60-68(2022)
DOI：

Yuan LIU, Yan-meng BI, Ting PAN, et al. Ethyl Acetate Fraction of Dicliptera chinensis (L.) Juss.Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathways. [J]. Chinese Journal of Integrative Medicine 28(1):60-68(2022) DOI： 10.1007/s11655-021-3298-5.

摘要

目的:

基于肝星状细胞模型及肝纤维化小鼠模型探讨狗肝菜乙酸乙酯提取物抗肝纤维化的分子机制.

方法:

体外实验中

首先将肝星状细胞用PDGF预处理以确保激活

并用狗肝菜乙酸乙酯提取物分别处理24和48小时; 采用CCK8、EdU染色、电镜、免疫荧光染色和蛋白印迹等方法分别评估狗肝菜乙酸乙酯提取物对肝星状细胞的作用. 体内实验中

通过持续6周腹腔注射四氯化碳 (2μL/g

25%浓度

每周3次) 制备肝纤维化小鼠模型. 将40只8周雄性C57BL /6小鼠随机分为4组

包括对照组、模型组、阳性对照组 (秋水仙碱0.2 mg/kg) 和狗肝菜乙酸乙酯提取物 (0.5 g/kg) 组. 狗肝菜乙酸乙酯提取物和秋水仙碱组小鼠给予狗肝菜乙酸乙酯提取物 (0.5 g/kg) 或秋水仙碱 (0.2 mg/kg) 每日一次灌胃

持续6周; 对照组及模型组小鼠给予等量生理盐水灌胃. 通过生化检测和HE、Masson染色评估肝纤维化程度. 采用蛋白质印迹法测定α-平滑肌肌动蛋白 (α-SMA) 和微管相关蛋白轻链3B (LC3B) 的蛋白表达水平.

结果:

细胞模型和肝纤维化小鼠模型检测结果均显示

狗肝菜乙酸乙酯提取物可以抑制肝星状细胞活化

减轻α-SMA蛋白表达

促进 LC3-I到LC3Ⅱ的转换增加 (

0.05) . 病理组织学检查结果表明

模型组小鼠肝脏具有明显纤维化病理改变

狗肝菜乙酸乙酯提取物组可见肝脏胶原纤维的沉积减少. 巴弗洛霉素A1和雷帕霉素在肝星状细胞干预结果表明

抑制自噬会增强α-SMA蛋白表达水平 (

0.01) . 蛋白质印迹分析显示PI3K、p-PI3K、AKT、p-AKT、mTOR、p-mTOR及p-p70S6K水平均降低 (

0.05) .

结论:

狗肝菜乙酸乙酯提取物通过诱导自噬发挥抗肝纤维化活性

狗肝菜乙酸乙酯提取物可能是一种用于人类肝纤维化治疗的潜在药物.

Abstract

Objective:

To investigate the molecular mechanism underlying the anti-hepatic fibrosis activity of ethyl acetate fraction

Dicliptera chinensis

(L.) Juss. (EDC) in human hepatic stellate cells (HSCs)

in vitro

and in a carbon tetrachloride (CCl

)-induced hepatic fibrosis mouse model

in vivo

Methods:

For

in vitro

study

HSCs were pre-treated with platelet-derived growth factor (10 ng/mL) for 2 h to ensure activation and treated with EDC for 24 h and 48 h

respectively. The effect of EDC on HSCs was assessed using cell counting kit-8 assay

EdU staining

transmission electron microscopy

immunofluorescence staining

and Western blot

respectively. For

in vivo

experiments

mice were intraperitoneally injected with CCl

(2 μL/g

adjusted to a 25% concentration in olive oil)

3 times per week for 6 weeks

to develop a hepatic fibrosis model. Forty 8-week-old male C57BL/6 mice were divided into 4 groups using a random number table (

=10)

including control

model

positive control and EDC treatment groups. Mice in the EDC and colchicine groups were intragastrically administered EDC (0.5 g/kg) or colchicine (0.2 mg/kg) once per day for 6 weeks. Mice in the control and model groups received an equal volume of saline. Biochemical assays and histological examinations were used to assess liver damage. Protein expression levels of α-smooth muscle actin (α-SMA) and microtubule-associated protein light chain 3B (LC3B) were measured by Western blot.

Results:

EDC reduced pathological damage associated with liver fibrosis

downregulated the expression of α-SMA and upregulated the expression of LC3B (

0.05)

both in HSCs and the CCl

-induced liver fibrosis mouse model. The intervention of bafilomycin A1 and rapamycin in HSCs strongly supported the notion that inhibition of autophagy enhanced α-SMA protein expression levels (

0.01). The results also found that the levels of phosphoinositide (PI3K)

p-PI3K

AKT

p-AKT

mammalian target of rapamycin (mTOR)

p-mTOR

and p-p70S6K all decreased after EDC treatment (

0.05).

Conclusion:

EDC has anti-hepatic fibrosis activity by inducing autophagy and might be a potential drug to be further developed for human liver fibrosis therapy.

关键词

Keywords

Dicliptera chinensis (L.) Juss.autophagyliver fibrosishepatic stellate cellsChinese medicine

references

Sun J, Xie Q, Tan D, Ning Q, Niu J, Bai X, et al. The 104-week efficacy and safety of telbivudine-based optimization strategy in chronic hepatitis B patients: a randomized,controlled study. Hepatology 2014;59:1283-1292.

Friedman SL. Evolving challenges in hepatic fibrosis. Nat Rev Gastroenterol Hepatol 2010;7:425-436.

Rockey DC. Translating an understanding of the pathogenesis of hepatic fibrosis to novel therapies. Clin Gastroenterol Hepatol 2013;11:224-231.

Ye B, Zhang Y, He W. Advances in research on Dicliptera chinensis (L.) Juss. China Pract Med (Chin) 2011;6:235-237.

Xu D, He Y, Yang S, Li K, Yang X, Shen Z. Protective effect of Dicliptera chinensis (L.) Juss. Polysaccharide P2B on carbon tetrachloride-induced injury of liver cell line L-02.China Pharmacist (Chin) 2016;19:675-677.

Xu Y, Gao Y, Zhong M, Li J, Cao H, Huang S, et al.Isolation, characterization and bioactivities of the polysaccharides from Dicliptera chinensis (L.) Juss. Int J Biol Macromol 2017;101:603-611.

Xu Y, Zhang K, Zhong M, Gao Y, Duan X. Advances in research on chemical constituents and pharmacology of Dicliptera chinensis (L.) Juss. Chin Pharm (Chin)2015;34:4862-4864.

Zhang K, Zhu H, Gao Y. Research on active extracts of Dicliptera chinensis (L.) Juss. on liver protection. China J Chin Mater Med (Chin) 2010;35:497-498.

Zhang K, Gao Y, Zhong M, Xu Y, Li J, Chen Y, et al. Hepatoprotective effects of Dicliptera chinensis polysaccharides on dimethylnitrosamine-induced hepatic fibrosis rats and its underlying mechanism. J Ethnopharmacol 2016;179:38-44.

Zhang X, Zhang J, Jia L, Xiao S. Dicliptera Chinensis polysaccharides target TGF-beta/Smad pathway and inhibit stellate cells activation in rats with dimethylnitrosamine-induced hepatic fibrosis. Cell Mol Biol (Noisy-le-grand)2016;62:99-103.

Denton D, Nicolson S, Kumar S. Cell death by autophagy:facts and apparent artefacts. Cell Death Differ 2012;19:87-95.

Mizushima N, Levine B, Cuervo AM, Klionsky DJ.Autophagy fights disease through cellular self-digestion.Nature 2008;451:1069-1075.

Jiang P, Mizushima N. Autophagy and human diseases.Cell Res 2014;24:69-79.

San-Miguel B, Crespo I, Sánchez DI, González-Fernández B, Ortiz de Urbina JJ, Tuñón MJ, et al. Melatonin inhibits autophagy and endoplasmic reticulum stress in mice with carbon tetrachloride-induced fibrosis. J Pineal Res 2015;59:151-162.

Lee YA, Wallace MC, Friedman SL. Pathobiology of liver fibrosis: a translational success story. Gut 2015;64:830-841.

Liu Y, Bi Y, Mo C, Zeng T, Huang S, Gao L, et al. Betulinic acid attenuates liver fibrosis by inducing autophagy via the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway. J Nat Med 2019;73:179-189.

Zhong W, Gao L, Zhou Z, Lin H, Chen C, Huang P, et al.Indoleamine 2,3-dioxygenase 1 deficiency attenuates CCl4-induced fibrosis through Th17 cells down-regulation and tryptophan 2,3-dioxygenase compensation. Oncotarget 2017;8:40486-40500.

Mimche PN, Brady LM, Bray CF, Lee CM, Thapa M, King TP, et al. The receptor tyrosine kinase EphB2 promotes hepatic fibrosis in mice. Hepatology 2015;62:900-914.

Mo C, Xie S, Zhong W, Zeng T, Huang S, Lai Y, et al. Mutual antagonism between indoleamine 2,3-dioxygenase 1 and nuclear factor E2-related factor 2 regulates the maturation status of DCs in liver fibrosis. Free Radic Biol Med 2020;160:178-190.

Dong S, Chen Q, Song Y, Sun Y, Wei B, Li X, et al. Mechanisms of CCl4-induced liver fibrosis with combined transcriptomic and proteomic analysis. J Toxicol Sci 2016;41:561-572.

Török NJ. Recent advances in the pathogenesis and diagnosis of liver fibrosis. J Gastroenterol 2008;43:315-321.

Zhao C, She T, Wang L, Su Y, Qu L, Gao Y, et al.Daucosterol inhibits cancer cell proliferation by inducing autophagy through reactive oxygen species-dependent manner. Life Sci 2015;137:37-43.

Shafik AN, Khodeir MM, Gouda NA, Mahmoud ME.Improved antifibrotic effect of a combination of verapamil and silymarin in rat-induced liver fibrosis. Arab J Gastroenterol 2011;12:143-149.

Chen SR, Chen XP, Lu JJ, Wang Y, Wang YT. Potent natural products and herbal medicines for treating liver fibrosis. Chin Med 2015;10:7.

Cheng Y, Mai JY, Wang MF, Chen GF, Ping J.Antifibrotic effect of total flavonoids of Astmgali Radix on dimethylnitrosamine-induced liver cirrhosis in rats. Chin J Integr Med 2017;23:48-54.

Zhang K, Xu Q, Gao Y, Cao H, Lian Y, Li Z, et al.Polysaccharides from Dicliptera chinensis ameliorate liver disturbance by regulating TLR-4/NF-κB and AMPK/Nrf2 signalling pathways. J Cell Mol Med 2020;24:6397-6409.

Chen JW, Ni BB, Li B, Yang YH, Jiang SD, Jiang LS. The responses of autophagy and apoptosis to oxidative stress in nucleus pulposus cells: implications for disc degeneration.Cell Physiol Biochem 2014;34:1175-1189.

Schneider JL, Cuervo AM. Liver autophagy: much more than just taking out the trash. Nat Rev Gastroenterol Hepatol 2014;11:187-200.

Boyle KB, Randow F. The role of 'eat-me' signals and autophagy cargo receptors in innate immunity. Curr Opin Microbiol 2013;16:339-348.

Tanida I, Minematsu-Ikeguchi N, Ueno T, Kominami E.Lysosomal turnover, but not a cellular level, of endogenous LC3 is a marker for autophagy. Autophagy 2005;1:84-91.

El-Mihi KA, Kenawy HI, El-Karef A, Elsherbiny NM, Eissa LA. Naringin attenuates thioacetamide-induced liver fibrosis in rats through modulation of the PI3K/Akt pathway. Life Sci 2017;187:50-57.

Li J, Li X, Xu W, Wang S, Hu Z, Zhang Q, et al. Antifibrotic effects of luteolin on hepatic stellate cells and liver fibrosis by targeting AKT/mTOR/p70S6K and TGFbeta/Smad signalling pathways. Liver Int 2015;35:1222-1233.

Lin X, Li Y, Zhang X, Wei Y, Wen S, Lu Z, et al. Tormentic acid inhibits hepatic stellate cells activation via blocking PI3K/Akt/mTOR and NF-κB signalling pathways. Cell Biochem Funct 2020; doi: 10.1002/cbf.3564http://doi.org/10.1002/cbf.3564.

Fan S, Zhang B, Luan P, Gu B, Wan Q, Huang X, et al.PI3K/AKT/mTOR/p70S6K pathway is involved in Aβ25-35-induced autophagy. BioMed Res Int 2015;2015:161020.

Shinji S, Yukiko S, Yoko I, Sumihiro K, Takahiro F, Isei T, et al. Caffeine induces apoptosis by enhancement of autophagy via PI3K/Akt/mTOR/p70S6K inhibition.Autophagy 2011;7:176-187.

Hemmings BA, Restuccia DF. PI3K-PKB/Akt pathway. Cold Spring Harb Perspect Biol 2012;4:a11189.

Klionsky DJ, Meijer AJ, Codogno P. Autophagy and p70S6 kinase. Autophagy 2005;1:59-60.

Destefano MA, Jacinto E. Regulation of insulin receptor substrate-1 by mTORC2 (mammalian target of rapamycin complex 2). Biochem Soc Trans 2013;41:896-901.

Lin NY, Beyer C, Giessl A, Kireva T, Scholtysek C,Uderhardt S, et al. Autophagy regulates TNF alpha-mediated joint destruction in experimental arthritis. Ann Rheumat Dis 2013;72:761-768.

Hernández-Gea V, Hilscher M, Rozenfeld R, Lim MP,Nieto N, Werner S, et al. Endoplasmic reticulum stress induces fibrogenic activity in hepatic stellate cells through autophagy. J Hepatol 2013;59:98-104.

Lin CW, Zhang H, Li M, Xiong X, Chen X, Chen X, et al. Pharmacological promotion of autophagy alleviates steatosis and injury in alcoholic and non-alcoholic fatty liver conditions in mice. J Hepatol 2013;58:993-999.

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Institute of Liver Diseases, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine

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Ethyl Acetate Fraction of Dicliptera chinensis (L.) Juss.Ameliorates Liver Fibrosis by Inducing Autophagy via PI3K/AKT/mTOR/p70S6K Signaling Pathways

DOI：10.1007/s11655-021-3298-5

摘要

Abstract

关键词

Keywords

references