2017 Fiscal Year Research-status Report
Protective role of CYGB in prevention of liver fibrosis development in vitro and in vivo
| Project/Area Number |
16K19363
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| Research Institution | Osaka City University |
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Principal Investigator |
LE THUY 大阪市立大学, 大学院医学研究科, 特任助教 (10572175)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Keywords | cytoglobin / liver fibrosis / hepatic stellate cells |
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| Outline of Annual Research Achievements |
Cytoglobin (CYGB) was originally discovered from rat hepatic stellate cells (HSCs) in 2001 by our group, and is the fourth globin in mammals. Cygb deficient mice, Cygb overexpressing mice, and recombinant human CYGB protein were generated in our laboratory. The role of Cygb in anti-fibrosis and cancer were examined in different models of liver injuries including (1) chemical induced liver fibrosis using thioacetamide (TAA) treatment for 10 weeks; (2) carcinogen induced liver cancer using diethylnitrosamine (DEN) for 12 months; (3) diet induced non-alcoholic fatty liver diseases (NASH) using choline deficient amino acid define (CDAA) diet for 16 weeks. We found that in the absence of Cygb liver injuries were magnified in Cygb-KO mice. In contrast, Cygb overexpression protected mice from liver damage induced by different etiologies. Furthermore, we produced rhCYGB protein and found that rhCYGB possess antioxidant and peroxidase activities. Human hepatic stellate cells (HHSteC) under rhCYGB treatment showed suppression of fibrosis related genes such as Collagene 1A1, αSMA, TIMP-1, TGFβ1. In vivo model, rhCYGB administration in TAA-treated mice inhibited liver injuries, inflammation and fibrosis development. Thus, CYGB with ROS scavenger could play an important role for preventing from liver fibrosis
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
TAA model: Compared to WT mice, 10 week-TAA treated Cygb-TG showed markedly decreased inflammation and fibrosis development as indicated by (1) reduced accumulation of neutrophils in the liver (- 2 fold, p< 0.0001) together with specific down regulation of transcription level of Ccl-2 (- 2,4 folds, p < 0.001); (2) declined oxidative stress condition demonstrated by significant decreased expression of NOX-2 at mRNA level; (3) obviously inhibited fibrosis development as shown by decreased Sirius-red positive area, significantly decreased αSMA at protein and mRNA level, together with blocked mRNA level of Col1a1, TGFβ3. CDAA model: Choline deficient L-amino acid-defined diet (CDAA) and its control diet (choline-supplied L-amino acid-defined diet, CSAA) are administrated to WT, Cygb / , and Cygb-TG mice for 16 weeks. The results showed that after 16 weeks of CDAA treatment and or recovery, KO mice still exhibited most severe liver inflammation and fibrosis development and remaining after recovery. In contrast, TG mice showed less inflammation, fibrosis and well recover. rhCYGB production and application:We have produced human recombinant CYGB protein, and examined its potential anti-fibrotic therapy. Interestingly, we found that the rhCYGB binding to human HSCs cells, HHSteC, and supressed collagen production at protein and mRNA level with time and dose dependent.
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| Strategy for Future Research Activity |
1.In vitro study: using primary mouse hepatic stellate cells (HSC) and human HSC cell line to examine the mechanism action of CYGB in protect cells from oxidative stress and toxicity induced liver injuries.
2.Produce rhCYGB for application in vivo with several models including NASH model using CDAA diet, fibrosis model using TAA treatment, or cholestasis liver injury using 3,5-diethoxycar¬bonyl-1,4-dihydrocollidine (DDC) diet.
3.Molecular biology of liver fibrosis and pathological analysis
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| Causes of Carryover |
Micro array is necessary using large number of samples.
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Research Products
(10 results)
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[Journal Article] Polymorphisms in MICA, but not in DEPDC5, HCP5 or PNPLA3, are associated with chronic hepatitis C-related hepatocellular carcinoma.2017
Author(s)
1.Hai H, Tamori A, Thuy LTT, Yoshida K, Hagihara A, Kawamura E, Uchida-Kobayashi S, Morikawa H, Enomoto M, Murakami Y, Kawada N
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Journal Title
Scientific Reports
Volume: 7(1)
Pages: 11920
DOI
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Fibroblast growth factor 2 (FGF2) regulates cytoglobin expression and activation of human hepatic stellate cells via JNK signaling.2017
Author(s)
2.Sato-Matsubara M, Matsubara T, Daikoku A, Okina Y, Longato L, Rombouts K, Thuy LTT, Adachi J, Tomonaga T, Ikeda K, Yoshizato K, Pinzani M, Kawada N.
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Journal Title
J Biol Chem.
Volume: 292(46)
Pages: 18961-18972
DOI
Peer Reviewed / Int'l Joint Research
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[Journal Article] The Hemoglobin Homolog Cytoglobin in Smooth Muscle Inhibits Apoptosis and Regulates Vascular Remodeling2017
Author(s)
3.Jourd'heuil FL, Xu H, Reilly T, McKellar K, El Alaoui C, Steppich J, Liu YF, Zhao W, Ginnan R, Conti D, Lopez-Soler R, Asif A, Keller RK, Schwarz JJ, Thanh Thuy LT, Kawada N, Long X, Singer HA, Jourd'heuil D.
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Journal Title
Arterioscler Thromb Vasc Biol.
Volume: 37(10)
Pages: 1944-1955
DOI
Peer Reviewed / Int'l Joint Research
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[Journal Article] Cytoglobin Regulates Blood Pressure and Vascular Tone through Nitric Oxide Metabolism in the Vascular Wall.2017
Author(s)
4.Xiaoping Liu, Mohamed A. El-Mahdy, James Boslett, Saradhadevi Varadharaj, Craig, Hemann, Tamer M. Abdelghany, Raed S. Ismail, Sean C. Little, Danlei Zhou, Le Thi Thanh Thuy, Norifumi Kawada, and Jay L. Zweier.
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Journal Title
Nature Communications.
Volume: 8
Pages: 14807
DOI
Peer Reviewed / Int'l Joint Research
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