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1.
Di Yi Jun Yi Da Xue Xue Bao ; 23(8): 820-2, 2003 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-12919908

RESUMO

OBJECTIVE: To find a new method for obtaining abundant, well-purified and functionally active rat islet cells cultured in vitro. METHOD: The pancreatic tissues of suckling rats underwent repeated digestion for short durations with collagenase, and the cell growth was observed under inverted microscope 18 h after cell inoculation. The cultured cells were then transferred to a new culture plate and the supernate was harvested regularly to determine the concentration of insulin, amylase and glucose-stimulated insulin release. RESULTS: The fibroblast cells in the primary cultured suckling rat cells were obviously reduced, and the ratio of dithizone-stained cells were 85%-90% with the viability exceeding 90% as assessed by trypan blue staining. The secretion function of the cultured cells remained normal after a 7- to 11-day culture. CONCLUSION: Repeated digestion of the pancreatic tissues for short durations with collagenase and timely cell transfer to new plate may help achieve highly purified viable monolayer islet cells that are well applicable for experimental studies.


Assuntos
Ilhotas Pancreáticas/citologia , Animais , Animais Lactentes , Separação Celular , Células Cultivadas , Imuno-Histoquímica , Insulina/análise , Ilhotas Pancreáticas/química , Ilhotas Pancreáticas/metabolismo , Ratos , Ratos Sprague-Dawley
2.
Di Yi Jun Yi Da Xue Xue Bao ; 23(6): 608-10, 2003 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-12810390

RESUMO

OBJECTIVE: To study the effect of triglycerides (TGs) on the function of in vitro cultured rat islet cells. METHODS: SD rat islet cells were isolated for monolayer cell culture and treated with TGs at different concentrations (0, 0.5, 2.5, 5.0, 10.0 mmol/L) for 72 h. Insulin level in the cell culture media was measured after glucose loading test, and the deposition of fat droplets in the islet cells observed by oil red O-staining. RESULTS: No obvious effect was observed after a 72-h incubation of the islet cells with TGs at low glucose level (2.8 mmol/L), while with the stimulation of high-level glucose (27.8 mmol/L), the insulin secretion was significantly inhibited by TGs of higher concentrations (5.0, 10.0 mmol/L). The deposition of fat droplets was found in the islet cells after incubation with TGs, with TGs of higher concentrations. CONCLUSION: TGs induces deposition of fat droplets in islet cells in vitro, and may inhibit the insulin-secreting function of the cells.


Assuntos
Ilhotas Pancreáticas/efeitos dos fármacos , Triglicerídeos/farmacologia , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Insulina/metabolismo , Secreção de Insulina , Ilhotas Pancreáticas/patologia , Ilhotas Pancreáticas/fisiologia , Metabolismo dos Lipídeos , Masculino , Ratos , Ratos Sprague-Dawley
3.
Di Yi Jun Yi Da Xue Xue Bao ; 23(5): 449-51, 2003 May.
Artigo em Chinês | MEDLINE | ID: mdl-12754126

RESUMO

OBJECTIVE: To investigate the in vitro apoptosis-inducing effect of palmitic acid on pancreatic islet cells in primary culture, thereby to understand the role of palmitic acid in the pathogenesis of type 2 diabetes mellitus. METHOD: SD rat pancreatic islet cells were isolated and cultured in monolayer in vitro followed by incubation with stepwise diluted palmitic acid (0, 0.125, 0.25 mmol/L and 0.5 mmol/L respectively), and the insulin concentrations in the culture medium were determined by radio immunological methods. Morphological observation with a fluorescence microscope was conducted after double staining of the cells with PI/Hoechst 33342. RESULTS: The glucose-stimulated insulin secretion was inhibited by palmitic acid at the concentration of 0.25 and 0.5 mmol/L, which also induced obvious cell apoptosis. CONCLUSION: Palmitic acid is capable of inducing islet cell apoptosis in a dose-dependent manner.


Assuntos
Apoptose/efeitos dos fármacos , Diabetes Mellitus Tipo 2/etiologia , Ilhotas Pancreáticas/efeitos dos fármacos , Ácido Palmítico/toxicidade , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Insulina/metabolismo , Secreção de Insulina , Ilhotas Pancreáticas/patologia , Necrose , Ratos , Ratos Sprague-Dawley
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