2016 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 | Hepatic stellate cells / fibrosis / cholestasis / cytoglobin |
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| Outline of Annual Research Achievements |
Bile duct ligation model (BDL): WT, Cygb / , and Cygb-TG mice at 9-10 weeks old are performed BDL or sham operation for acute phase (24, 48, and 72 hours), and chronic phase (1-3 weeks). BDL-TG mice showed the most protective liver injury compared to WT/KO. Toxic model: Thioacetamid（TAA）model in WT, Cygb / , and Cygb-TG mice are performed. Mice are injected intraperitoneally (i.p.) with escalation dose of TAA, twice a week, start with 50 μg/g body weight (BW) until 400 μg/g BW in 10 weeks. Control mice are injected with saline. TAA-TG liver showed decreased transcriptional TNFα, IL-6, chemokines including Ccl2. Generation of human recombinant CYGB (rhCYGB) protein is performed. Human Cygb cDNA is cloned into the pRSETA vector. CYGB protein is further purified and heme assay was confirmed. Human HSCs cells treated with rhCYGB reduced αSMA expression.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
We aim to (1) assess the effect of Cygb-overexpressing in HSCs on the development of liver fibrosis; (2) explore the mechanism action of Cygb including its impact on the oxidative stress and anti-oxidant system which in turn inhibits HSC activation and prevents fibrosis formation; (3) produce and examine the therapeutic approach of human recombinant CYGB in preventing liver fibrosis.
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| Strategy for Future Research Activity |
In vivo application of rhCYGB protein
CYGB protein is also assessed in vivo to find the effects of fibrosis regression using the above designed models of liver fibrosis including i) fibrotic model group (given TAA only); ii) TAA + low dose rhCYGB; iii) TAA + high dose rhCYGB. The dose of rhCYGB will be selected based on our pilot studies. In both in vitro and in vivo model, we evaluate the effect of CYGB on the growth of SCs exposed to various concentrations of CYGB protein. Cell viability, cell cycle arrest, HSC apoptosis, expression of profibrotic mediators and ECM degradation modulators in HSCs are examined. Especially, the up-stream and down-stream targets of CYGB are of our goal to establish the signalling pathway of CYGB in the regression of fibrosis.
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| Causes of Carryover |
I have attended two international meeting in 2016, in Hamburg and in Boston.
XIXth International conference on "Oxygen binding and sensing proteins", 2016, September 11-14, University of Hamburg, Hamburg, Germany. I have learnt new information of proteins in the globin family, in which Cytoglobin is a novel member.The American Association for the Study of Liver Diseases (AASLD) 2016, November 11-15, Boston, USA. This meeting is the biggest meeting in the field of hepatology in the world. Attending this meeting gave me a lots of information, technique, and hint for my research.
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| Expenditure Plan for Carryover Budget |
The fiscal year 2017, I will use the amount of about 900.000 yen, including the materials, and reagents for my experiments, and attend the American Association for the Study of Liver Diseases (AASLD) 2017 in Washington DC, USA.
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Research Products
(6 results)
