Effective Protection on Acute Liver Injury by Halo Tag-Flanked Recombinant Fibroblast Growth Factor 7.

The drug development of FGF7 has been restricted by its toxicity to the host, low expression, poor stability, and easy degradation. Recent studies have shown that Halo-tag-flanked recombinant human FGF7 can solve the problem of toxicity; however, its biological activity is unknown. This study aimed to explore the activity of Halo-rhFGF7 and rhFGF7 on acute liver injury in vitro and in vivo. The rhFGF7 is expressed with a N-terminal Halo-tag, followed by a tobacco etch virus (TEV) protease cleavage site, in Escherichia coli BL21 (DE3) pLysS in this study. The products could stimulate the proliferation of carbon tetrachloride-damaged L-O2 cells (normal human liver cells); they also inhibited cell apoptosis. Due to the use of the Halo, the protein could be tracked using fluorescence localization. Recombinant protein exerted a protective effect on the acute liver injury model in vitro and in vivo. The MTT assay and Western blot analysis showed that this protective effect is realized through various paths, including promoting proliferation, inhibiting cell apoptosis and anti-inflammatory. In conclusion, Halo-rhFGF7 and rhFGF7 displayed an excellent protective effect on acute liver injury. The present study provided an experimental basis and data support for further research on rhFGF7.

[1]  I. Yoruk,et al.  An analysis on the roles of angiogenesis-related factors including serum vitamin D, soluble endoglin (sEng), soluble fms-like tyrosine kinase 1 (sFlt1), and vascular endothelial growth factor (VEGF) in the diagnosis and severity of late-onset preeclampsia , 2017, The journal of maternal-fetal & neonatal medicine : the official journal of the European Association of Perinatal Medicine, the Federation of Asia and Oceania Perinatal Societies, the International Society of Perinatal Obstetricians.

[2]  A. Lyubimov,et al.  Protected graft copolymer-formulated fibroblast growth factors mitigate the lethality of partial body irradiation injury , 2017, PloS one.

[3]  A. Tyagi,et al.  RETRACTED ARTICLE: Recombinant human keratinocyte growth factor attenuates apoptosis in elastase induced emphysematous mice lungs , 2017, Inhalation toxicology.

[4]  S. Kotnala,et al.  Recombinant Human Keratinocyte Growth Factor Induces Akt Mediated Cell Survival Progression in Emphysematous Mice. , 2015, Archivos de bronconeumologia.

[5]  X. Mao,et al.  HaloTag is an effective expression and solubilisation fusion partner for a range of fibroblast growth factors , 2015, PeerJ.

[6]  B. Liu,et al.  Recombinant Keratinocyte Growth Factor 1 in Tobacco Potentially Promotes Wound Healing in Diabetic Rats , 2014, BioMed research international.

[7]  T. L. Thuoc,et al.  An overview on keratinocyte growth factor: from the molecular properties to clinical applications. , 2014, Protein and peptide letters.

[8]  Xiaokun Li,et al.  Highly efficient expression of functional recombinant human keratinocyte growth factor 1 and its protective effects on hepatocytes , 2014, Applied Microbiology and Biotechnology.

[9]  K. Kwon,et al.  Send Orders of Reprints at Reprints@benthamscience.org the Halotag: Improving Soluble Expression and Applications in Protein Functional Analysis , 2022 .

[10]  Songwen Zhang,et al.  Fibroblast growth factor 7 inhibits cholesterol 7α-hydroxylase gene expression in hepatocytes. , 2012, Biochemical and biophysical research communications.

[11]  D. Van Tyne,et al.  Live Imaging of Mitosomes and Hydrogenosomes by HaloTag Technology , 2012, PloS one.

[12]  Su-Mei Tsai,et al.  Expression and Function of Fibroblast Growth Factor (FGF) 7 during Liver Regeneration , 2011, Cellular Physiology and Biochemistry.

[13]  S. Werner,et al.  FGF receptors 1 and 2 control chemically induced injury and compound detoxification in regenerating livers of mice. , 2010, Gastroenterology.

[14]  K. Wood,et al.  Methods for detection of protein-protein and protein-DNA interactions using HaloTag. , 2008, Methods in molecular biology.

[15]  C. Lang,et al.  HaloTag: a new versatile reporter gene system in plant cells. , 2006, Journal of experimental botany.

[16]  J. Rubin,et al.  Keratinocyte growth factor expression and activity in cancer: implications for use in patients with solid tumors. , 2006, Journal of the National Cancer Institute.

[17]  K. Patel.  Drug-related hepatotoxicity. , 2006, The New England journal of medicine.

[18]  M. Clark,et al.  Sudden Enlargement of the Neck , 2006 .

[19]  M. V. D. van den Brink,et al.  Regeneration Keratinocyte Growth Factor (KGF) Is Required For Post-Natal Thymic , 2011 .

[20]  Masashi Suzuki,et al.  Expression of fibroblast growth factors and their receptors during full-thickness skin wound healing in young and aged mice. , 2005, The Journal of endocrinology.

[21]  M. Goddard,et al.  Keratinocyte growth factor therapy in murine oleic acid-induced acute lung injury. , 2005, American journal of physiology. Lung cellular and molecular physiology.

[22]  P. Ray.  Protection of epithelial cells by keratinocyte growth factor signaling. , 2005, Proceedings of the American Thoracic Society.

[23]  D. Danilenko,et al.  Effect of Recombinant Human Keratinocyte Growth Factor (rHuKGF) on the Immunopathogenesis of Intestinal Graft-Vs.-Host Disease Induced Without a Preconditioning Regimen , 2004, Journal of Clinical Immunology.

[24]  J. Rubin,et al.  Keratinocyte growth factor/fibroblast growth factor 7, a homeostatic factor with therapeutic potential for epithelial protection and repair. , 2004, Advances in cancer research.

[25]  T. Arakawa,et al.  Stabilization of recombinant human keratinocyte growth factor by osmolytes and salts. , 1996, Journal of pharmaceutical sciences.

[26]  A. Diehl.  Keratinocyte growth factor induces proliferation of hepatocytes and epithelial cells throughout the rat gastrointestinal tract: Housley RM, Morris CF, Boyle W, Ring B, Biltz R, Tarpley JE, Aukerman SL, et al. J Clin Invest 1994; 94: 1764–1777 , 1995 .