| Project/Area Number |
08671147
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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 |
内分泌・代謝学
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| Research Institution | SHIGA UNIVERSITY OF MEDICAL SCIENCE |
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Principal Investigator |
HANEDA Masakazu SHIGA UNIBERSITY OF MEDICAL SCIENCE,THIRD DEPARTMENT OF MEDICINE,ASSISTANT PROFESSOR, 医学部, 助手 (60164894)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIO Yoshihiko SHIGA UNIVERSITY OF MEDICAL SCIENCE,THIRD DEPARTMENT OF MEDICINE,ASSISTANT PROFE, 医学部, 助手 (40281084)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Diabetic nephropathy / Mesangial cells / Advanced glycation end products(AGE) / mRNA differential display / gene expression |
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| Research Abstract |
Nonenzymatic glycation is proposed to be one of the etiologic factors of diabetic nephropathy. Although receptors for advanced glycation end products (AGEs) were found in glomerular mesangial cells, AGEs-induced changes in cellular functions have not been clarified yet. We performed mRNA differential display analysis, to identify AGEs-induced alterations of gene expression in cultured rat mesangial cells. Confluent mesangial cells were incubated with AGE-BSA (200mug/ml) or control BSA (200mug/ml) for 48 hours and total RNA was extracted. After reverse transcription with T12MN primers, cDNA was amplified by polymerase chain reaction using T12MN primers and random 10 mer, and the products were analyzed on a sequence gel. Of 6,000 mRNAs screened, 78 candidate PCR products were detected on a sequence gel. Northen blot analysis confirmed 6 clones changed by AGEs. Three of them were increased by AGEs and three clones were decreased. Two of three clones increased by AGEs showed homology to mitochondrial elongation factor Tu and mesenchyme fork head-1, respectively. One of three clones decreased by AGEs showed homology to actin depolymerizing factor. These three genes have been reported to be involved in protein synthesis or cell proliferation. Further analysis of these genes may provide new insight on pathogenesis of diabetic nephropathy. In conclusion, three genes regulated by AGEs in mesangial cells were identified by mRNA differential display analysis.
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