Attenuation of Expression of γ-Glutamylcysteine Synthetase by Ribozyme Transfection Enhance Insulin Secretion by Pancreatic β Cell Line, MIN6
| Autor: | Hideaki Kondo, Hironori Yamasaki, Takahiro Abe, Hiroshi Kijima, Hirofumi Takino, Takahito Kondo, Yasunori Sera, Yoshihiko Yamaguchi, Yoshishige Urata, Eiji Kawasaki, Satoko Mori, Kenji Yamakawa, Katsumi Eguchi, Iida Tetsuya, Masako Ozaki |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Antioxidant
Glutamate-Cysteine Ligase medicine.medical_treatment Biophysics Enzyme-Linked Immunosorbent Assay Transfection Biochemistry Cell Line Islets of Langerhans Mice chemistry.chemical_compound Adenosine Triphosphate Insulin Secretion medicine Animals Insulin RNA Catalytic RNA Messenger Molecular Biology Fluorescent Dyes chemistry.chemical_classification Aniline Compounds Dose-Response Relationship Drug biology Ribozyme Cell Biology Glutathione Blotting Northern Molecular biology Cell biology Glucose Enzyme Microscopy Fluorescence Xanthenes chemistry Cell culture biology.protein Calcium Signal transduction Intracellular Plasmids |
| Zdroj: | Biochemical and Biophysical Research Communications. 278:236-240 |
| ISSN: | 0006-291X |
| DOI: | 10.1006/bbrc.2000.3776 |
| Popis: | Low levels of intracellular antioxidant enzyme activities as well as glutathione (GSH) concentrations have been described in pancreatic beta cells. We examined the effects of intracellular GSH depletion on insulin secretion and the role of intracellular GSH in signal transduction in beta cell line, MIN6 cells. Anti-gamma-glutamylcysteine synthetase (gamma-GCS) heavy subunit ribozyme was stably transfected to MIN6 cells to reduce intracellular GSH concentration. In the presence of 10 mM glucose, ribozyme-transfected cells (RTC) increased insulin secretion from 0.58 microg/10(6) cells/h in control cells (CC) to 1.48 microg/10(6) cells/h. This was associated with increased intracellular Ca(2+) concentration in RTC, detected by fluo-3 staining. Our results demonstrated that intracellular GSH concentration might influence insulin secretion by MIN6 cells, and suggest that enhanced insulin secretion by beta cells conditioned by chronic depletion of GSH is mediated by increased intracellular Ca(2+) concentration. |
| Databáze: | OpenAIRE |
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