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KMID : 0377220000250020029
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Medical Journal of Chosun Univercity
2000 Volume.25 No. 2 p.29 ~ p.36
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Effects of high glucose on the suoeroxide dismutase gene expression in rat insulinoma cell line
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Lee Jae-Woo
Lee Byoung-Rai
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Abstract
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Background: Reactive oxygen species are produced under diabetic condition and possibly cause various forms of tissue damage in patients with diabetes mellitus. The aim of this study is to examine the effects of high glucose on the gene expression of superoxide dismutase(SOD) isozymes in rat insulinoma cells line.
Methods: Rat insulinoma(RINm5F) cells were cultured with RPMI 1640 media containing high glucose(22.2mM), or normo-glucose(5.6mM) for 3 days and for 21 days. The SOD activities were determined by spectrophotometric assay and nitroblue tetrazolium (NBT) stain. The levels of SOD mRNA were quantified by reverse transcriptase-polymerase chain reaction (RT-PCR).
Results: The CuZn-SOD activity was decreased but Mn- SOD activity was increased significantly in RINm5F cells which cultured with high glucose(22.2mM) media for 3 days. In RINm5F cells, the Mn-SOD mRNA content was also increased by high glucose, but the level of CuZn-SOD mRNA was not changed by gluose concentration for 3 days culture. Both CuZn-SOD and Mn- SOD activity were increased significantly when cultured with high glucose media (22.2mM) for 21 days. The levels of Mn-SOD and CuZn-SOD mRNA were increased in RINm5F cells cultured with in high glucose media for 21 days.
Conclusion: These results suggest that in RINm5F cells, both CuZn-SOD and Mn-SOD gene expressions maybe increase by high glucose and decrement of CuZn-SOD activity, in cells of short term culture with high glucose contained media may due to the protein modification.
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KEYWORD
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SOD, High glucose, Insulinoma cell
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