| Project/Area Number |
12672206
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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 |
応用薬理学・医療系薬学
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| Research Institution | TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY |
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Principal Investigator |
HOSOYA Ken-ichi Toyama Medical and Pharmaceutical University Faculty of Pharmaceutical Sciences Professor, 薬学部, 教授 (70301033)
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| Co-Investigator(Kenkyū-buntansha) |
KATAYAMA Kazunori Toyama Medical and Pharmaceutical University Faculty of Pharmaceutical Sciences Associate Professor, 薬学部, 助教授 (70126514)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | inner blood-retinal barrier / retinal endothelial cell line / retinal pericyte cell line / cell growth regulating factor / conditionally immortalized cell line / co-culture / system xc / oxidative stress condition / TGFβ1 / 条件的不死化細胞株 / 液性因子 / 血管新生 / system x_c^- |
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| Research Abstract |
Conditionally immortalized rat retinal capillary endothelial cell line (TR-iBRB) and retinal pericyte cell line (TR-rPCT) were established from transgenic rats harboring the temperature-sensitive simian virus 40 large T-antigen gene. The paracrine interaction between TR-iBRB and TR-rPCT cells has been investigated to elucidate the cell growth regulating factor in pericyte. TR-iBRB cells were seeded on the insert of Transwell chamber and TR-rPCT cell were seeded under the insert (contact co-culture model) and on the well chamber (non-contact co-culture model). The conditioned medium for TR-rPCT cells was also used to observe the cell growth of TR-iBRB cells. The growth of TR-iBRB cells was significantly slowed down with increasing concentration of TR-rPCT conditioned medium. In contact co-culture model, the growth of TR-iBRB cells was arrested 4 days after the beginning of the co-culture. These results suggest that some factor released from pericyte cells is involved in regulating the cell growth in retinal capillary endothelial cells.
The [^<14>C] L-cystine uptake by TR-iBRB cells appeared to be mediated through a saturable Na^+-independent process with a Michaelis-Menten constant of 9.2 μM, suggesting system xc, which consists of XCT and 4F2hc mRNA, mediated L-cystine uptake. After 100 μM diethyl maleate treatment, which is a model agent for oxidative stress, the XCT mRNA level and L-cystine uptake activity in TR-iBRB cells were enhanced in a time-dependent manner. Concomitantly, the glutathione concentration in TR-iBRB cells was increased. In contrast, the 4F2hc mRNA level was unchanged up to 24 hours and was induced for more than 24 hours by DEM pretreatment. These findings suggest L-cystine transport is activated by induction of system xc under the oxidative stress conditions at the inner blood-retinal barrier.
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