Skip to main content
SpringerLink
Log in

Caffeine alleviates acute liver injury by inducing the expression of NEDD4L and deceasing GRP78 level via ubiquitination

  • Original Research Article
  • Published:
Inflammation Research Aims and scope Submit manuscript

Abstract

Background

Acute liver injury is liver cell injury that occurs rapidly in a short period of time. Caffeine has been shown to maintain hepatoprotective effect with an unclear mechanism. Endoplasmic reticulum stress (ERS) has significant effects in acute liver injury. Induction of GRP78 is a hallmark of ERS. Whether or not caffeine’s function is related to GRP78 remains to be explored.

Methods

Acute liver injury model was established by LPS-treated L02 cells and in vivo administration of LPS/D-Gal in mice. Caffeine was pre-treated in L02 cells or mice. Gene levels was determined by real-time PCR and western blot. Cell viability was tested by CCK-8 assay and cell apoptosis was tested by flow cytometry. The interaction of GRP78 and NEDD4L was determined by Pull-down and co-immunoprecipitation (Co-IP) assay. The ubiquitination by NEDD4L on GRP78 was validated by in vitro ubiquitination assay.

Results

Caffeine protected liver cells against acute injury induced cell apoptosis and ERS both in vitro and in vivo. Suppression of GRP78 could block the LPS-induced cell apoptosis and ERS. NEDD4L was found to interact with GRP78 and ubiquitinate its lysine of 324 site directly. Caffeine treatment induced the expression of NEDD4L, resulting in the ubiquitination and inhibition of GRP78.

Conclusion

Caffeine mitigated the acute liver injury by stimulating NEDD4L expression, which inhibited GRP78 expression via ubiquitination at its K324 site. Low dose of caffeine could be a promising therapeutic treatment for acute liver injury.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data availability

All data generated or analyzed during this study are included in this article. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

ER:

Endoplasmic reticulum

ERS:

Endoplasmic reticulum stress

GRP78:

Glucose-regulated protein 78

PERK:

Protein kinase R-like ER kinase

IRE-1α:

Inositol-requiring enzyme 1 alpha

ATF6:

Activating transcription factor 6

ATF4:

Activating transcription factor 6

CHOP:

C/EBP homologous protein

CAFF:

Caffeine

H and E:

Hematoxylin–eosin

FBS:

Fetal bovine serum

qRT-PCR:

Quantitative real-time PCR

CCK8:

Cell counting kit-8

LPS:

Lipopolysaccharide

D-gal:

D-galactosamine

Co-IP:

Co-immunoprecipitation

References

  1. Ding W, Fan YY, Zhang C, Fu L, Chen X, Xu DX. Obeticholic acid differentially regulates hepatic injury and inflammation at different stages of D-galactosamine/lipopolysaccharide-evoked acute liver failure. Eur J Pharm. 2019;850:150–7.

    Article  CAS  Google Scholar 

  2. Huang ZB, Zheng YX, Li N, Cai SL, Huang Y, Wang J, et al. Protective effects of specific cannabinoid receptor 2 agonist GW405833 on concanavalin A-induced acute liver injury in mice. Acta Pharmacol Sin. 2019;40:1404–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Alpert L, Hart J. The pathology of alcoholic liver disease. Clin Liver Dis. 2016;20:473–89.

    Article  PubMed  Google Scholar 

  4. Clark VC, Marek G, Liu C, Collinsworth A, Shuster J, Kurtz T, et al. Clinical and histologic features of adults with alpha-1 antitrypsin deficiency in a non-cirrhotic cohort. J Hepatol. 2018;69:1357–64.

    Article  CAS  PubMed  Google Scholar 

  5. Kroemer G, Galluzzi L, Vandenabeele P, Abrams J, Alnemri ES, Baehrecke EH, et al. Classification of cell death: recommendations of the nomenclature committee on cell death 2009. Cell Death Differ. 2009;16:3–11.

    Article  CAS  PubMed  Google Scholar 

  6. Mahdi AA, Rizvi SH, Parveen A. Role of endoplasmic reticulum stress and unfolded protein responses in health and diseases. Indian J Clin Biochem. 2016;31:127–37.

    Article  CAS  PubMed  Google Scholar 

  7. Iurlaro R, Munoz-Pinedo C. Cell death induced by endoplasmic reticulum stress. FEBS J. 2016;283:2640–52.

    Article  CAS  PubMed  Google Scholar 

  8. Liu G, Sun Y, Li Z, Song T, Wang H, Zhang Y, et al. Apoptosis induced by endoplasmic reticulum stress involved in diabetic kidney disease. Biochem Biophys Res Commun. 2008;370:651–6.

    Article  CAS  PubMed  Google Scholar 

  9. Bernales S, Papa FR, Walter P. Intracellular signaling by the unfolded protein response. Annu Rev Cell Dev Biol. 2006;22:487–508.

    Article  CAS  PubMed  Google Scholar 

  10. Lien YC, Kung HN, Lu KS, Jeng CJ, Chau YP. Involvement of endoplasmic reticulum stress and activation of MAP kinases in beta-lapachone-induced human prostate cancer cell apoptosis. Histol Histopathol. 2008;23:1299–308.

    CAS  PubMed  Google Scholar 

  11. Harada N, Okuyama M, Yoshikatsu A, Yamamoto H, Ishiwata S, Hamada C, et al. Endoplasmic reticulum stress in mice increases hepatic expression of genes carrying a premature termination codon via a nutritional status-independent GRP78-dependent mechanism. J Cell Biochem. 2017;118:3810–24.

    Article  CAS  PubMed  Google Scholar 

  12. Wang X, Wu X, Wang Q, Zhang Y, Wang C, Chen J. NLRP6 suppresses gastric cancer growth via GRP78 ubiquitination. Exp Cell Res. 2020;395: 112177.

    Article  CAS  PubMed  Google Scholar 

  13. Hughes A, Oxford AE, Tawara K, Jorcyk CL, Oxford JT. Endoplasmic reticulum stress and unfolded protein response in cartilage pathophysiology; contributing factors to apoptosis and osteoarthritis. Int J Mol Sci. 2017;18:665.

    Article  PubMed Central  Google Scholar 

  14. Meng X, Zhu Y, Tao L, Zhao S, Qiu S. Periostin has a protective role in melatonin-induced cell apoptosis by inhibiting the eIF2alphaATF4 pathway in human osteoblasts. Int J Mol Med. 2018;41:1003–12.

    CAS  PubMed  Google Scholar 

  15. Tian RD, Chen YQ, He YH, Tang YJ, Chen GM, Yang FW, et al. Phosphorylation of eIF2alpha mitigates endoplasmic reticulum stress and hepatocyte necroptosis in acute liver injury. Ann Hepatol. 2020;19:79–87.

    Article  CAS  PubMed  Google Scholar 

  16. Ruhl CE, Everhart JE. Coffee and tea consumption are associated with a lower incidence of chronic liver disease in the United States. Gastroenterology. 2005;129:1928–36.

    Article  PubMed  Google Scholar 

  17. Azam S, Hadi N, Khan NU, Hadi SM. Antioxidant and prooxidant properties of caffeine, theobromine and xanthine. Med Sci Monit. 2003;9:BR325–30.

    CAS  PubMed  Google Scholar 

  18. Cachon AU, Quintal-Novelo C, Medina-Escobedo G, Castro-Aguilar G, Moo-Puc RE. Hepatoprotective effect of low doses of caffeine on CCl4-induced liver damage in rats. J Diet Suppl. 2017;14:158–72.

    Article  CAS  PubMed  Google Scholar 

  19. Yang B, Kumar S. Nedd4 and Nedd4-2: closely related ubiquitin-protein ligases with distinct physiological functions. Cell Death Differ. 2010;17:68–77.

    Article  CAS  PubMed  Google Scholar 

  20. Kwak YD, Wang B, Li JJ, Wang R, Deng Q, Diao S, et al. Upregulation of the E3 ligase NEDD4-1 by oxidative stress degrades IGF-1 receptor protein in neurodegeneration. J Neurosci. 2012;32:10971–81.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Wang H, Sun RQ, Camera D, Zeng XY, Jo E, Chan SM, et al. Endoplasmic reticulum stress up-regulates Nedd4-2 to induce autophagy. FASEB J. 2016;30:2549–56.

    Article  PubMed  Google Scholar 

  22. Zhou YP, Xia Q. Inhibition of miR-103a-3p suppresses lipopolysaccharide-induced sepsis and liver injury by regulating FBXW7 expression. Cell Biol Int. 2020;44:1798–810.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Li Y, Chen Y, Huang H, Shi M, Yang W, Kuang J, et al. Autophagy mediated by endoplasmic reticulum stress enhances the caffeine-induced apoptosis of hepatic stellate cells. Int J Mol Med. 2017;40:1405–14.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Hu J, Zhu Z, Ying H, Yao J, Ma H, Li L, et al. Oleoylethanolamide protects against acute liver injury by regulating Nrf-2/HO-1 and NLRP3 pathways in mice. Front Pharm. 2020;11: 605065.

    Article  CAS  Google Scholar 

  25. Li S, Jin S, Chen W, Yu J, Fang P, Zhou G, et al. Mangiferin alleviates endoplasmic reticulum stress in acute liver injury by regulating the miR-20a/miR-101a-Nrf2 axis. J Biochem. 2020;168:365–74.

    Article  CAS  PubMed  Google Scholar 

  26. Pan Y, Long X, Yi R, Zhao X. Polyphenols in Liu Bao tea can prevent CCl(4)-induced hepatic damage in mice through its antioxidant capacities. Nutrients. 2018;10:1280.

    Article  PubMed Central  Google Scholar 

  27. Shi T, Song W, Xu R. Autophagy and ER stress in LPS/GalN induced acute liver injury. Mol Med Rep. 2017;16:7001–5.

    Article  CAS  PubMed  Google Scholar 

  28. Tai M, Zhang J, Song S, Miao R, Liu S, Pang Q, et al. Protective effects of luteolin against acetaminophen-induced acute liver failure in mouse. Int Immunopharmacol. 2015;27:164–70.

    Article  CAS  PubMed  Google Scholar 

  29. Clarke R, Cook KL, Hu R, Facey CO, Tavassoly I, Schwartz JL, et al. Endoplasmic reticulum stress, the unfolded protein response, autophagy, and the integrated regulation of breast cancer cell fate. Cancer Res. 2012;72:1321–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Clarke R, Shajahan AN, Wang Y, Tyson JJ, Riggins RB, Weiner LM, et al. Endoplasmic reticulum stress, the unfolded protein response, and gene network modeling in antiestrogen resistant breast cancer. Horm Mol Biol Clin Investig. 2011;5:35–44.

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Hu R, Warri A, Jin L, Zwart A, Riggins RB, Fang HB, et al. NF-kappaB signaling is required for XBP1 (unspliced and spliced)-mediated effects on antiestrogen responsiveness and cell fate decisions in breast cancer. Mol Cell Biol. 2015;35:379–90.

    Article  PubMed  Google Scholar 

  32. Cook KL, Soto-Pantoja DR, Clarke PA, Cruz MI, Zwart A, Warri A, et al. Endoplasmic reticulum stress protein GRP78 modulates lipid metabolism to control drug sensitivity and antitumor immunity in breast cancer. Cancer Res. 2016;76:5657–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Chang YJ, Huang YP, Li ZL, Chen CH. GRP78 knockdown enhances apoptosis via the down-regulation of oxidative stress and Akt pathway after epirubicin treatment in colon cancer DLD-1 cells. PLoS ONE. 2012;7: e35123.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Samanta S, Yang S, Debnath B, Xue D, Kuang Y, Ramkumar K, et al. The Hydroxyquinoline analogue YUM70 inhibits GRP78 to induce ER stress-mediated apoptosis in pancreatic cancer. Cancer Res. 2021;81:1883–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Wu J, Wu Y, Lian X. Targeted inhibition of GRP78 by HA15 promotes apoptosis of lung cancer cells accompanied by ER stress and autophagy. Biol Open. 2020;9:bio053298.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Mukwevho E, Kohn TA, Lang D, Nyatia E, Smith J, Ojuka EO. Caffeine induces hyperacetylation of histones at the MEF2 site on the Glut4 promoter and increases MEF2A binding to the site via a CaMK-dependent mechanism. Am J Physiol Endocrinol Metab. 2008;294:E582–8.

    Article  CAS  PubMed  Google Scholar 

  37. Ping J, Wang JF, Liu L, Yan YE, Liu F, Lei YY, et al. Prenatal caffeine ingestion induces aberrant DNA methylation and histone acetylation of steroidogenic factor 1 and inhibits fetal adrenal steroidogenesis. Toxicology. 2014;321:53–61.

    Article  CAS  PubMed  Google Scholar 

  38. Yu H, Yang T, Gao P, Wei X, Zhang H, Xiong S, et al. Caffeine intake antagonizes salt sensitive hypertension through improvement of renal sodium handling. Sci Rep. 2016;6:25746.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Kawaratani H, Tsujimoto T, Douhara A, Takaya H, Moriya K, Namisaki T, et al. The effect of inflammatory cytokines in alcoholic liver disease. Mediators Inflamm. 2013;2013: 495156.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Su GL. Lipopolysaccharides in liver injury: molecular mechanisms of kupffer cell activation. Am J Physiol Gastrointest Liver Physiol. 2002;283:G256–65.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments.

Funding

This work was supported by grant from National Natural Sciences Foundation of China (No.81970523), and Natural Sciences Foundation of Hunan province (2020JJ4877).

Author information

Authors and Affiliations

  1. Department of Emergency, Xiangya Hospital Central South University, Changsha, 410008, Hunan Province, People’s Republic of China

    Xing-Wang Hu & Ying-Hui Xiong

  2. Department of Infectious Diseases/Hunan Provincial Key Laboratory of Viral Hepatitis, Xiangya Hospital Central South University, Changsha, 410008, Hunan Province, People’s Republic of China

    Xiang-Min Li, Ai-Min Wang, Fang-Jie Zhang, Feng Zeng, Li-Ping Cao, Hui Long, Ji Xu & Jia Li

  3. Scientific Research Center, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou, 510080, Guangdong Province, People’s Republic of China

    Yong-Ming Fu

Authors
  1. Xing-Wang Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiang-Min Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ai-Min Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yong-Ming Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fang-Jie Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Feng Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Li-Ping Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hui Long
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ying-Hui Xiong
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Ji Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Jia Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

XWH: conceptualization, writing–original draft, and methodology; XML: formal analysis; AMW: supervision; YMF: validation; FJZ: data curation; FZ: resources; LPC: investigation; HL: software; YHX: visualization; JX: project administration; JL: funding acquisition, writing–review and editing. All authors have read and approved the final version of this manuscript to be published.

Corresponding author

Correspondence to Jia Li.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethical approval

Not applicable. This article does not contain any studies with human participants or animals performed by any of the authors.

Consent for publication

Not applicable. This article does not contain any studies with human participants performed by any of the authors.

Additional information

Responsible Editor: John Di Battista.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

11_2022_1603_MOESM1_ESM.tif

Supplementary file1 NEDD4L inhibited liver cell apoptosis via regulating GRP78. A mRNA and B protein levels of NEDD4L and GRP78 in L02 cells. C Flow cytometry and statistic result for apoptosis detection of L02 cells. *P < 0.05, **P < 0.01, ***P < 0.001, one-way ANOVA followed by Tukey’s multiple comparison test, N = 3. (TIF 1122 KB)

11_2022_1603_MOESM2_ESM.tif

Supplementary file2 The effect of caffeine treatment on LPS level. Relative level of LPS in the culture medium of L02 cells after treated with or without caffeine (TIF 100 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hu, XW., Li, XM., Wang, AM. et al. Caffeine alleviates acute liver injury by inducing the expression of NEDD4L and deceasing GRP78 level via ubiquitination. Inflamm. Res. 71, 1213–1227 (2022). https://doi.org/10.1007/s00011-022-01603-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00011-022-01603-0

Keywords

Search

Navigation