| Project/Area Number |
17592160
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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 |
Periodontal dentistry
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| Research Institution | Health Sciences University of Hokkaido |
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Principal Investigator |
NAKASHIMA Keisuke Health Science University of Hokkaido, Periodontology and Endodontology, Associate Professor, 歯学部, 助教授 (80227785)
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| Co-Investigator(Kenkyū-buntansha) |
KATO Satsuki Health Science University of Hokkaido, Periodontology and Endodontology, Lecturer, 歯学部, 講師 (50281283)
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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 |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Cell / Tissue / Dent is t ry / Bio-molecule / Protein |
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| Research Abstract |
Four peptides, hLF33 (GRRRR SVQWC AVSQP EATKC FQWQR NMRKV RGP), hLF33K (GRRRR SVQWC AVSQP EATKC FQWQR NMKKV RGP), hLF20-37 (C FQWQR NMRKV RGPPV SC) and hLF20-37S (C FQWQR NMRKV RGPPV SC; the S-S bond between cysteine residues) were prepared, and measured for inhibitory effects against the LPS-LBP binding. Effects of these peptides were one-tenth of hLF. No significant differences were observed among the peptides at the concentration of 2.5 μm. However, the rank order of effects for these peptides was as follows at the concentrations of 100 μM: hLf33 >> hLf33K >> hLf20-37 > hLf20-37S. These results suggest that the 28th amino acid from the N-terminal of hLF is involved in inhibitory effects against the LPS-LBP binding, while the S-S bond between cysteine residues does not. These peptides also inhibited TNF-α production, however, inhibitory effects of the peptides were much less than hLF.
We further performed chemotaxis assay with 96-well plates. THP-1 cells were preincubated in the presence of la, 25-Dihydroxyvitamin D3 for 18 h. The cells were transferred on the filter of ChemoTx 96-well disposable chamber: the lower chamber contained each chemoattractants in the presence or absence of 200 μg/m1 of hLF. Plates were centrifuged, and the number of cells that migrated to the lower chamber was determined with AQueous cell proliferation assay kit. The number of migrated cells to LPS was significantly lower than the controls. Inhibition of cell migration was observed at concentrations > or = 10 ng/ml of LPS, and the inhibition was partially abrogated by the addition of hLF in a dose-dependent manner. The results suggest that LPS inhibits random migration of THP-1 cells, and that hLF protects against the inhibition of cell migration. hLF may regulate cell migration in an inflammatory response to periodontopathic bacteria.
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