| Project/Area Number |
21K14812
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 38050:Food sciences-related
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| Research Institution | Hokkaido University |
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Principal Investigator |
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| Project Period (FY) |
2021-04-01 – 2025-03-31
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| Project Status |
Granted (Fiscal Year 2023)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2024: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2023: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2022: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
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| Keywords | Hijiki / obesity / Sphingolipid metabolism / lipids / LC/MS / Obesity |
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| Outline of Research at the Start |
(1) Isolate and characterize the chemical constituent of Hijiki responsible for SMS inhibition. (2) Determination of its IC50 and mode of action on the lipid droplet formation by cell-based assay (3) Revealing the in vitro effects by analysis of altered cellular lipid biomarkers by LC-MS (4) Elucidate the in vivo effects by oral administration of the active constituent of
Hijiki using High-fat diet mouse model.
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| Outline of Annual Research Achievements |
We achieved significant progress in our research in the last year. We successfully established the sphingomylein synthase (SMS) monitoring assay method using LC/MS and validated by in-vitro and in-silico studies using a known inhibitor for SMS. Also, the manuscript was prepared and submitted to the international peer-reviewed journal. We successfully identified the compound in Hijiki responsible for SMSs nhibition by mass spectrometry and NMR techniques. The identified compound was confirmed with authentic standard. Further, we investigated the inhibition affinity of active compounds with SMS enzymes by docking studies. The summary of results including isolation characterization and SMS assay results are under preparation for a possible manuscript while further studies are in progress.
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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
Due to the characterization of the active component in Hijiki, there is slight delay in the progress, however it is going rather smoothly now as we successfully characterized the active component of hijiki responsible for SMS inhibition.
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| Strategy for Future Research Activity |
We have now isolated the active component of Hijiki with complete characterization by spectroscopic and spectrometry techniques. Our future goal is to test this active compound in in-vitro and in-vivo system. We will first perform the in-vitro activity of the Hijiki derived SMS inhibitor using fatty liver disease cell model by LC/MS and fluorescence microscopy techniques. The inhibition at cellular levels will be confirmed by measuring sphingomylein lipid in the C3A cells. Based on the in-vitro results, the in-vivo studies will be conducted as planned in the proposal and an overall report will be prepared by the end of the fiscal year. Additionally, the manuscript will be prepared to submit to an international journal and also results will be presented at a domestic conference.
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