2002 Fiscal Year Final Research Report Summary
targeting of iNOS with antisense DNA plasmid prevents cytokine-induced inhibition of osteoblastic differentiation
| Project/Area Number |
13672082
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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 |
Surgical dentistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
HIKIJI Hisako Tokyo University, Health Service Centre, Assistant professor, 保健管理センター, 講師 (50292876)
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| Co-Investigator(Kenkyū-buntansha) |
TOYO-OKA Teruhiko Tokyo University, Faculty of medicine. Professor, 医学部附属病院, 所長 教授 (00146151)
TAKATO Tuyoshi Tokyo University, Faculty of medicine, Professor, 医学部附属病院, 教授 (90171454)
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| Project Period (FY) |
2001 – 2002
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| Keywords | iNOS / antisenae / gene targeting / gene therapy / bone destruction |
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| Research Abstract |
Purpose :
Proinflammatry cytokines, tumor necrosis factor (TNF) -α combined with interleukin-1 (IL-1) -β, induce excessive production of nitric oxide (NO) and its cytotoxic metabolite, peroxynitrite (ONOO^-) via indueible nitric oxide synthase (iNOS) in mouse osteoblasts. In this investigation, to estimate the effects of antisense DNA of iNOS on osteoblastic activity, we produced transformed cell lines with antisense plasmid targeting the iNOS gene for long-lasting inhibition The identifications of the transformed antisense cell lines were 1) the detection of antisense transcripts, 2) the attenuated expression of iNOS protein, 3) the reduction of NOS activity and 4) the level of NO production. These cell lines targeting which showed decreased production of both NO and ONOO^-, prevented the inhibition iNOS, of osteoblastic differentiation as was assayed by the mRNA expression of alkaline phosphatase (ALPase) in the presence of proinflammatory cytokines. Present results indicate that the antisense DNA plasmid of iNOS is potent to prevent the cytokine-induced reduction of osteoblastic activity.
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