| Project/Area Number |
17590071
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | KAGAWA NUTRITION UNIVERSITY |
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Principal Investigator |
HORIE SHUICHI KAGAWA NUTRITION UNIVERSITY, DEPT. OF CLINICAL BIOCHEMISTRY, PROFESSOR, 栄養学部, 教授 (60157063)
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| Co-Investigator(Kenkyū-buntansha) |
OHKURA NAOKI TEIKYO UNIV., FAC.PHARM. SCI., DEPT. OF CLINICAL MOLECULAR BIOLOGY, LECTURER, 薬学部, 専任講師 (60349256)
OISHI KATSUTAKA NATL. INST. ADV. IND. SCI. AND TEC., INSTI. BIOL. RESOURCES AND FUNC., SENIOR RESEARCHER, 生物機能工学研究部門, 研究員 (50338688)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2005: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | BIOLOGICAL RHYTHM / TIME RELATED GENES / PERIPHERAL CLOCKS / CIRCADIAN RHYTHM / VASUCLAR ENDOTHELIAL CELL / THROMBOMODULIN / ANTH-THROMBOSIS / NUCLEAR CLOCK MOLECULE |
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| Research Abstract |
In the present study, we checked whether or not the endothelial thrombomodulin (TM) expression is controlled by peripheral clock genes in vivo and in vitro and obtained following results.
1) Under the condition of constant light-dark cycle, northern blot analysis of mouse lung showed a circadian variation of TM mRNA level, and the expression rhythms were obviously damped in Clock mutant mice.
2) The restricted feeding shifted the circadian phase of TM mRNA expression in the lung of wild mice.
3) When cultured human microvascular endothelial cells were exposed to serum shock, a temporal fluctuation of TM gene expression was observed.
4) A gel mobility shift analysis showed that the components of nuclear extracts from the cells interact with distal E-box in the TM gene, and in vitro translation product of CLOCK bound to the E-box in the presence of that of BMAL-2, but not of BMAL-1.
5) Co-transfection of expression plasmids of CLOCK and BMAL-2 in COS-7 cells increased the promoter activity of reporter plasmid containing distal but not proximal E-box sequence, and the activation by CLOCK/BMAL-2 was reduced in the presence of another clock regulatory protein, CRY.
6) When cultured human endothelial cells were exposed to serum shock, a temporal fluctuation of binding activity between the nuclear proteins and the E-box sequence was observed in a gel mobility shift experiment.
These results show that TM expression was under the control of peripheral clocks such as CLOCK and BMAL-2, and suggest that the disorder of TM expression on the vascular surface on the vascular surface by irregularity of peripheral clock oscillations which might be caused by un-regulatory food supplement may brought to the various thrombotic diseases.
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