Iron enhances hepatic fibrogenesis and activates TGF-β signaling in murine hepatic stellate cells

Mehta, K.J., Coombes, J.D., Briones-Orta, M., Manka, P.P., Williams, R., Patel, V.B. and Sin, W.-K. 2018. Iron enhances hepatic fibrogenesis and activates TGF-β signaling in murine hepatic stellate cells. American Journal of the Medical Sciences. 355 (2), pp. 183-190.

TitleIron enhances hepatic fibrogenesis and activates TGF-β signaling in murine hepatic stellate cells
AuthorsMehta, K.J., Coombes, J.D., Briones-Orta, M., Manka, P.P., Williams, R., Patel, V.B. and Sin, W.-K.
Abstract

Introduction

Although excess iron induces oxidative stress in the liver, it is unclear whether it directly activates the hepatic stellate cells (HSC).

Methods

We evaluated effects of excess iron on fibrogenesis and TGF-β signaling in murine HSC. Cells were treated with holotransferrin (0.005–5 g/L) for 24 h, with or without the iron chelator deferoxamine (10 µM). Gene expressions (α-SMA, Col1-α1, Serpine-1, TGF-β, Hif1-α, Tfrc and Slc40a1) were analyzed by qRT-PCR, while TfR1, ferroportin, ferritin, vimentin, collagen, TGF-β RII and phospho-Smad2 proteins were evaluated by immunofluorescence, western blot and ELISA.

Results

HSC express the iron-uptake protein TfR1, and the iron-export protein ferroportin. Holotransferrin up-regulated TfR1 expression by 1.8-fold (p<0.03) and ferritin accumulation (iron storage) by 2-fold (p<0.01), and activated HSC with 2-fold elevations (p<0.03) in α-SMA mRNA and collagen secretion, and a 1.6-fold increase (p<0.01) in vimentin protein. Moreover, holotransferrin activated the TGF-β pathway with TGF-β mRNA elevated 1.6-fold (p=0.05), and protein levels of TGF-β RII and phospho-Smad2 increased by 1.8-fold (p<0.01) and 1.6-fold (p<0.01), respectively. By contrast, iron chelation decreased ferritin levels by 30% (p<0.03), inhibited collagen secretion by 60% (p<0.01), repressed fibrogenic genes α-SMA (0.2-fold; p<0.05) and TGF-β (0.4-fold; p<0.01), and reduced levels of TGF-β RII and phospho-Smad2 proteins.

Conclusion

HSC express iron transport proteins. Holotransferrin (iron) activates HSC fibrogenesis and the TGF-β pathway, while iron depletion by chelation reverses this, suggesting that this could be a useful adjunct therapy for patients with fibrosis. Further studies in primary human HSC and animal models are necessary to confirm this.

KeywordsFibrosis, Fibroblasts, Holotransferrin, Liver
JournalAmerican Journal of the Medical Sciences
Journal citation355 (2), pp. 183-190
ISSN0002-9629
Year2018
PublisherElsevier
Accepted author manuscriptMehta et al Revised Figures.pdf
Supplementary data or filesRevised Supplementary data 25.1.2017.pdf
Mehta et al Revised Figures.pdf
Digital Object Identifier (DOI)doi:10.1016/j.amjms.2017.08.012
Publication dates
Published in printFeb 2018
Published online23 Aug 2017
Published23 Aug 2017
LicenseCC BY-NC-ND 4.0

Related outputs

Excessive iron accumulation induces beta cell cellular oxidative stress and insulin secretory dysfunction in MIN6 cells
Blesia, V., Patel, V., Al‐Obaidi, H. and Zariwala, M. 2019. Excessive iron accumulation induces beta cell cellular oxidative stress and insulin secretory dysfunction in MIN6 cells. Diabetes UK Professional Conference 2019. Liverpool 06 - 08 Mar 2019 Wiley.

Mucosa-associated invariant T cells link intestinal immunity with antibacterial immune defects in alcoholic liver disease
Riva, A., Patel, V., Kurioka, A., Jeffery, H.C., Wright, G., Tarff, S., Shawcross, D., Ryan, J.M., Evans, A., Azarian, S., Bajaj, J.S., Fagan, A., Patel, V., Mehta, K., Lopez, C., Simonova, M., Katzarov, K., Hadzhiolova, T., Pavlova, S., Wendon, J.A., Oo, Y.T., Klenerman, P., Williams, R. and Chokshi, S. 2018. Mucosa-associated invariant T cells link intestinal immunity with antibacterial immune defects in alcoholic liver disease. Gut. 67 (5), pp. 918-930.

Faecal Calprotectin and a Twenty-Four-Parameter Questionnaire in Autistic Children with Gastrointestinal Symptoms
Bramati-castellarin, I., Patel, V.B. and Drysdale, I.P. 2017. Faecal Calprotectin and a Twenty-Four-Parameter Questionnaire in Autistic Children with Gastrointestinal Symptoms. SM Journal of Psychiatry & Mental Health. 2 (1), p. 1009.

HFE mRNA expression is responsive to intracellular and extracellular iron loading
Mehta, K., Farnaud, S. and Patel, V.B. 2017. HFE mRNA expression is responsive to intracellular and extracellular iron loading. Molecular Biology Reports. 44 (5), pp. 399-403.

Alcohol metabolism: implications for nutrution and health
Patel, V.B. and Preedy, V.R. 2017. Alcohol metabolism: implications for nutrution and health. in: Geissler, C. and Powers, H. (ed.) Human Nutrition, 13th Edition Oxford Oxford University Press. pp. 216-235

Iron enhances hepatic fibrogenesis through TGF-β activation
Mehta, K., Briones-Orta, M., Manka P., Coombes, J.D., Williams, R., Patel, V.B. and Syn, W-K. 2016. Iron enhances hepatic fibrogenesis through TGF-β activation. Hepatology. 64 (1 suppl), p. 360A.

Characterization of hepcidin response to holotransferrin in novel recombinant TfR1 HepG2 cells
Mehta, K., Busbridge, Mark, Renshaw, D., Evans, R.W., Farnaud, S. and Patel, V. 2016. Characterization of hepcidin response to holotransferrin in novel recombinant TfR1 HepG2 cells. Blood Cells, Molecules, and Diseases. 61, pp. 37-45.

Repeat-measures longitudinal study evaluating behavioural and gastrointestinal symptoms in children with autism before, during and after visceral osteopathic technique (VOT)
Bramati-Castellarin, I., Patel, V. and Drysdale, I.P. 2016. Repeat-measures longitudinal study evaluating behavioural and gastrointestinal symptoms in children with autism before, during and after visceral osteopathic technique (VOT). Journal of Bodywork and Movement Therapies. 20 (3), pp. 461-470.

Biomarkers in Health and Disease: Further Knowledge
Rajendram, R., Rajendram, R., Patel, V.B. and Preedy, V.R. 2015. Biomarkers in Health and Disease: Further Knowledge. in: Preedy, V.R. and Patel, V.B. (ed.) General Methods in Biomarker Research and their Applications Netherlands Springer. pp. 1311-1316

Regional Increase in the Expression of the BCAT Proteins in Alzheimer's Disease Brain: Implications in Glutamate Toxicity
Hull, J., Patel, V.B., El Hindy, M., Hezwani, M., Love, S., Kehoe, P.G., Chalmers, C. and Conway, M.E. 2015. Regional Increase in the Expression of the BCAT Proteins in Alzheimer's Disease Brain: Implications in Glutamate Toxicity. Journal of Alzheimer's Disease. 45 (3), pp. 891-905.

New insights into the role of the branched-chain aminotransferase proteins in the human brain
Hull, J., Patel, V.B., Hutson, S.M. and Conway, M.E. 2015. New insights into the role of the branched-chain aminotransferase proteins in the human brain. Journal of Neuroscience Research. 93, pp. 987-998.

Characterisation of hepcidin response to holotransferrin treatment in CHO TRVb-1 cells
Mehta, K., Greenwell, P., Renshaw, D., Busbridge, Mark, Garcia, M, Farnaud, S. and Patel, V. B. 2015. Characterisation of hepcidin response to holotransferrin treatment in CHO TRVb-1 cells. Blood Cells, Molecules, and Diseases. 55 (2), pp. 110-118.

Betaine, in context
Mehta, K. and Patel, V. 2015. Betaine, in context. in: Preedy, V.R. (ed.) Betaine: chemistry, analysis, function and effects Royal Society of Chemistry.

Cardiovascular disease in aging and the role of oxidative stress
Alzaid, F., Patel, V. and Preedy, V.R. 2014. Cardiovascular disease in aging and the role of oxidative stress. in: Preedy, V.R. (ed.) Aging: oxidative stress and dietary antioxidants Academic Press. pp. 23-36

The branched-chain aminotransferase proteins: novel redox chaperones for protein disulfide isomerase: implications in Alzheimer’s disease
Conway, M.E., El Hindy, M., Hezwani, M., Hull, J., Corry, D., Mansbridge, A., Hassler, M., Patel, V., Kehoe, P.G., Love, S., Harris, M., Forshaw, T., Lee, C.S., El Amraoui, F. and Wilson, A. 2014. The branched-chain aminotransferase proteins: novel redox chaperones for protein disulfide isomerase: implications in Alzheimer’s disease. Antioxidants and Redox Signaling. 20 (16), pp. 2497-2513.

Hepcidin transcription and peptide secretion are governed by different regulatory mechanisms
Mehta, K., Patel, V., Busbridge, M., Renshaw, D., Zariwala, M., Evans, R. and Farnaud, S. 2013. Hepcidin transcription and peptide secretion are governed by different regulatory mechanisms. Fifth Congress of the International BioIron Society (IBIS), Biennial World Meeting. University College London 14 - 18 Apr 2013 Wiley.

Interlinking diet, nutrition, menopause and recommended resources
Rajendram, R., Rajendram, R., Patel, V. and Preedy, V.R. 2013. Interlinking diet, nutrition, menopause and recommended resources. in: Hollins Martin, C.J., Watson, R.R. and Preedy, V.R. (ed.) Nutrition and diet in menopause Springer. pp. 451-456

Effect of Salvia miltiorrhiza on antioxidant enzymes in diabetic patients
Quan, Q., Quan, S. and Patel, V. 2013. Effect of Salvia miltiorrhiza on antioxidant enzymes in diabetic patients. in: Preedy, V.R. (ed.) Diabetes: oxidative stress and dietary antioxidants Academic Press. pp. 79-87

Hepcidin transcription and peptide secretion are governed by different regulatory mechanisms
Mehta, K., Patel, V., Busbridge Mark, Renshaw, D., Zariwala, M.G., Evans, R. and Farnaud, S. 2013. Hepcidin transcription and peptide secretion are governed by different regulatory mechanisms. American Journal of Hematology. 88, p. E55.

The role of cancer stem cells in osteosarcoma
Jenks, A., Smith, C.L., Patel, V. and Clements, M.O. 2013. The role of cancer stem cells in osteosarcoma. International society for stem cell research conference. Boston (Massachusetts, USA) June 12 – 15 2013

Damaging effect of free fatty acids and alcohol on hepatic mitochondrial function
Gyamfi, D., Clemens, D.L. and Patel, V. 2013. Damaging effect of free fatty acids and alcohol on hepatic mitochondrial function. Alcohol & Alcoholism. 48, pp. i48-i50.

Evaluation of behavioural and gastrointestinal symptoms in autistic children after visceral osteopathic treatment
Castellarin, I.B., Drysdale, I. and Patel, V. 2013. Evaluation of behavioural and gastrointestinal symptoms in autistic children after visceral osteopathic treatment. International Journal of Osteopathy. 16, pp. e13-e14.

Hepatic mitochondrial dysfunction induced by fatty acids and ethanol
Gyamfi, D., Everitt, H.E., Tewfik, I., Clemens, D.L. and Patel, V. 2012. Hepatic mitochondrial dysfunction induced by fatty acids and ethanol. Free Radical Biology & Medicine. 53 (11), pp. 2131-2145.

Increased transferrin-bound iron uptake in HepG2 cells is followed by an increase in expression of the iron hormone hepcidin pre-and post-translationally
Mehta, K., Patel, V., Busbridge Mark, Renshaw, D., Zariwala, M., Evans, R. and Farnaud, S. 2012. Increased transferrin-bound iron uptake in HepG2 cells is followed by an increase in expression of the iron hormone hepcidin pre-and post-translationally. European Iron Club Annual Meeting 2012. Rennes, France 29 - 31 Aug 2012

Chinese herbal products in the prevention and treatment of liver disease
Gyamfi, D., Everitt, H.E. and Patel, V. 2012. Chinese herbal products in the prevention and treatment of liver disease. in: Watson, R.R. and Preedy, V.R. (ed.) Bioactive food as dietary interventions for liver and gastrointestinal disease Academic Press. pp. 537-550

Dietary sugars: TLC screening of sugars in urine and blood samples
Alonso-Fernandez, J.R. and Patel, V. 2012. Dietary sugars: TLC screening of sugars in urine and blood samples. in: Preedy, V.R. (ed.) Dietary sugars: chemistry, analysis, function and effects Royal Society of Chemistry. pp. 186-205

Alterations to liver energy metabolism and cell death following alcohol exposure
Gyamfi, D., Clemens, D.L. and Patel, V. 2011. Alterations to liver energy metabolism and cell death following alcohol exposure. Alcohol & Alcoholism. 46, pp. p47-48.

Apoptosis in alcoholic liver disease
Gyamfi, D., Everitt, H.E. and Patel, V. 2010. Apoptosis in alcoholic liver disease. in: Preedy, V.R. (ed.) Apoptosis: modern insights into disease from molecules to man Science Publishers. pp. 211-233

The effect of alcohol exposure on liver energy metabolism and cell death
Gyamfi, D., Clemens, D.L. and Patel, V. 2010. The effect of alcohol exposure on liver energy metabolism and cell death. Hepatology. 52, p. 455A.

S-Nitrosoglutathione inactivation of the mitochondrial and cytosolic BCAT proteins: S-Nitrosation and S-Thiolation
Coles, S.J., Easton, P., Sharrod, H., Hutson, S.M., Hancock, J., Patel, V. and Conway, M.E. 2009. S-Nitrosoglutathione inactivation of the mitochondrial and cytosolic BCAT proteins: S-Nitrosation and S-Thiolation. Biochemistry. 48 (3), pp. 645-656.

Liver metabolism: biochemical and molecular regulations
Gyamfi, D. and Patel, V. 2009. Liver metabolism: biochemical and molecular regulations. in: Preedy, V.R. (ed.) Nutrition, diet therapy, and the liver London CRC Press. pp. 3-15

Hepatic oxidative stress and apoptosis following acute alcohol
Everitt, H.E., Tewfik, I., Preedy, V.R. and Patel, V. 2009. Hepatic oxidative stress and apoptosis following acute alcohol. Alcohol & Alcoholism. 44, p. p85.

Tomatoes: origin, cultivation techniques, and germplasm resources
da Silva, D.J.H., Abreu, F.B., Caliman, F.R.B., Antonio, A.C. and Patel, V. 2008. Tomatoes: origin, cultivation techniques, and germplasm resources. in: Preedy, V.R. and Watson, R.R. (ed.) Tomatoes and tomato products: nutritional, medicinal and therapeutic properties CRC Press. pp. 3-26

Effects of 4-hydroxynonenal on mitochondrial 3-hydroxy-3methylglutaryl (HMG-CoA) synthase
Patel, V., Spencer, C.H., Young, T.A., Lively, M.O. and Cunningham, C.C. 2007. Effects of 4-hydroxynonenal on mitochondrial 3-hydroxy-3methylglutaryl (HMG-CoA) synthase. Free Radical Biology and Medicine. 43 (11), pp. 1499-1507.

Nutrition and alcoholic liver disease
Everitt, H.E., Patel, V. and Tewfik, I. 2007. Nutrition and alcoholic liver disease. Nutrition Bulletin. 32 (2), pp. 138-144.

Protein adduct species in muscle and liver of rats following acute ethanol administration
Patel, V., Worrall, S., Emery, P.W. and Preedy, V.R. 2005. Protein adduct species in muscle and liver of rats following acute ethanol administration. Alcohol & Alcoholism. 40 (6), pp. 485-493.

Liver dysfunction induced by bile duct ligation and galactosamine injection alters cardiac protein synthesis
Hunter, R.V., Patel, V., Baker, A.J. and Preedy, V.R. 2004. Liver dysfunction induced by bile duct ligation and galactosamine injection alters cardiac protein synthesis. Metabolism: Clinical and Experimental. 53 (8), pp. 964-968.

Protein metabolism in alcohol misuse and toxicity
Preedy, V.R., Koll, M., Adachi, J., Mantle, D., Patel, V. and Peters, T. 2003. Protein metabolism in alcohol misuse and toxicity. in: Watson, R.R. and Preedy, V.R. (ed.) Nutrition and alcohol: linking nutrient interactions and dietary intake London CRC Press. pp. 261-300

Chronic ethanol consumption increases the concentration of a 4-hydroxynonenal adduct with the mitochondrial hydroxy methyl glutaryl CoA synthase in liver
Patel, V., Spencer, C. and Cunningham, C.C. 2003. Chronic ethanol consumption increases the concentration of a 4-hydroxynonenal adduct with the mitochondrial hydroxy methyl glutaryl CoA synthase in liver. Hepatology. 38, p. p393.

Emerging techniques in biomedical research and their application to alcohol toxicity
Patel, V., Chaurand, P., Caprioli, R., Austen, B., Frears, E., Manca, F., Davies, H., Vrana, K., Wheeler, M. and Preedy, V.R. 2003. Emerging techniques in biomedical research and their application to alcohol toxicity. Alcoholism: Clinical and Experimental Research. 27 (2), pp. 348-353.

Altered hepatic mitochondrial ribosome structure following chronic ethanol consumption
Patel, V. and Cunningham, C.C. 2002. Altered hepatic mitochondrial ribosome structure following chronic ethanol consumption. Archives of Biochemistry and Biophysics. 398 (1), pp. 41-50.

Chronic ethanol consumption increases the formation of an aldehyde-protein adduct in hepatic mitochondria
Patel, V., Young, T.A. and Cunningham, C.C. 2002. Chronic ethanol consumption increases the formation of an aldehyde-protein adduct in hepatic mitochondria. Alcoholism: Clinical and Experimental Research.

Cardioprotective effect of propranolol from alcohol-induced heart muscle damage as assessed by plasma cardiac troponin-T.
Patel, V., Ajmal, R., Sherwood, R., Sullivan, A., Richardson, P. and Preedy, V.R. 2001. Cardioprotective effect of propranolol from alcohol-induced heart muscle damage as assessed by plasma cardiac troponin-T. Alcoholism: Clinical and Experimental Research. 25 (6), pp. 882-889.

Diarrhea reduces the rates of cardiac protein synthesis in myofibrillar protein fractions in rats in vivo
Hunter, R.V., Patel, V., Miell, J., Wong, H.J., Marway, J., Richardson, P. and Preedy, V.R. 2001. Diarrhea reduces the rates of cardiac protein synthesis in myofibrillar protein fractions in rats in vivo. Journal of Nutrition. 131 (5), pp. 1513-1519.

Acute doxorubicin (adriamycin) dosage does not reduce cardiac protein synthesis in vivo, but decreases diaminopeptidase I and proline endopeptidase activities
Zima, T., Tesar, V., Mantle, D., Koll, M., Patel, V., Richardson, P. and Preedy, V.R. 2001. Acute doxorubicin (adriamycin) dosage does not reduce cardiac protein synthesis in vivo, but decreases diaminopeptidase I and proline endopeptidase activities. Experimental and Molecular Pathology. 70 (2), pp. 154-161.

Physiochemical properties of rat liver mitochondrial ribosomes
Patel, V., Cunningham, C.C. and Hantgan, R. 2001. Physiochemical properties of rat liver mitochondrial ribosomes. Journal of Biological Chemistry. 276 (9), pp. 6739-6746.

In vivo protein synthetic rates of atrial, ventricular, and pulmonary tissue proteins in aortic constriction, goldblatt, and bromoethylamine models of hypertension
Siddiq, T., Patel, V., Sherwood, R., Richardson, P. and Preedy, V.R. 2001. In vivo protein synthetic rates of atrial, ventricular, and pulmonary tissue proteins in aortic constriction, goldblatt, and bromoethylamine models of hypertension. Experimental and Molecular Pathology. 70 (1), pp. 19-30.

Ethanol and oxidative stress
Sun, A., Ingelman-Sundberg, M., Neve, E., Matsumoto, H., Nishitani, Y., Minowa, Y., Fukui, Y., Bailey, S., Patel, V., Cunningham, C.C., Zima, T., Fialova, L., Mikulikova, L., Popov, P., Malbohan, I., Janebova, M., Nespor, K. and Sun, G. 2001. Ethanol and oxidative stress. Alcoholism: Clinical and Experimental Research. 25 (5 Suppl.), pp. 237S-243S.

Alpha-Tocopherol supplementation does not prevent acute alcohol-induced heart muscle damage
Koll, M., Patel, V., Sherwood, R.A., Seitz, H.K., Simanowski, U.A., Richardson, P.J., Peters, T.J. and Preedy, V.R. 2001. Alpha-Tocopherol supplementation does not prevent acute alcohol-induced heart muscle damage. Proceedings of the Nutrition Society. 60, p. P125A.

Ethanol and protein metabolism
Cunningham, C.C., Preedy, V.R., Paice, A., Hesketh, J., Peters, T., Patel, V., Volpi, E., Mawatari, K., Masaki, H., Mori, H. and Torii, K. 2001. Ethanol and protein metabolism. Alcoholism: Clinical and Experimental Research. 25 (5 Suppl.), pp. 262S-268S.

Chronic ethanol consumption alters the glutathione/glutathione peroxidase-1 system and protein oxidation status in rat liver
Bailey, S., Patel, V., Young, T.A., Asayama, K. and Cunningham, C.C. 2001. Chronic ethanol consumption alters the glutathione/glutathione peroxidase-1 system and protein oxidation status in rat liver. Alcoholism: Clinical and Experimental Research. 25 (5), pp. 726-733.

Permalink - https://westminsterresearch.westminster.ac.uk/item/q20wv/iron-enhances-hepatic-fibrogenesis-and-activates-tgf-signaling-in-murine-hepatic-stellate-cells


Share this
Tweet
Email