RESUMEN
Amiodarone perturbs thyroid function, causing overt hypothyroidism or hyperthyroidism in 15% of patients. Changes in thyroid function are likely due, at least in part, to amiodarone and/or desethylamiodarone (DEA) concentration into the thyroid gland, but mechanisms whereby the drug uptake occurred are not known. Thyroidal (FRTL-5) or non-thyroidal [Chinese hamster ovary wild-type (CHOwt) or CHO stably transfected with NIS (CHO-NIS)] cells were exposed to 10 microM amiodarone or DEA. Cellular content of both drugs was measured by HPLC and normalized by protein concentration. Cellular concentration of the two drugs was higher in FRTL-5 (mean +/- SD 17.2 +/- 1.2 microg/mg protein of amiodarone and 18.9 +/- 0.7 microg/mg protein of DEA) than in CHO-NIS and CHOwt cells (10.8 +/- 0.8 microg/mg protein and 12.8 +/- 0.2 microg/mg protein, respectively, of amiodarone (p < 0.004); 11.9 +/- 0.1 microg/mg protein and 11 +/- 0.2 microg/mg protein, respectively, of DEA (p < 0.0002). DEA concentration was higher than that of amiodarone in all cell lines (p < 0.002). Differences between FRTL-5 and CHO cell lines were not dependent on TSH: in fact, cellular content of either drug did not change in the presence or absence of TSH in the culture medium. NIS did not intervene in amiodarone or DEA entry into thyroid cells, since amiodarone and DEA content was the same in CHOwt and CHO-NIS cells; in addition, KClO4 inhibited NIS function, but had no effect on drug uptake by the cells. At variance, 80 microM DEA reduced 125I uptake by CHO-NIS cells by 40% at 30 min without affecting cell viability. In conclusion, mechanisms whereby amiodarone is taken up by thyroid cells remain largely unknown, but the two main factors involved in thyroid-specific cellular transport, ie, NIS and TSH, seem to be excluded.
Asunto(s)
Amiodarona/farmacocinética , Glándula Tiroides/metabolismo , Amiodarona/análogos & derivados , Amiodarona/farmacología , Animales , Células CHO , Línea Celular , Cricetinae , Cricetulus , Yodo/farmacocinética , Percloratos/farmacología , Compuestos de Potasio/farmacología , Simportadores/genética , Glándula Tiroides/efectos de los fármacos , TransfecciónRESUMEN
OBJECTIVE: Chinese hamster ovary (CHO) cells transfected with human engineered insulin receptor (IR) cDNA to mutate Cys 860 to Ser (CHO-IR(C860S)) showed a defective insulin internalization without affecting insulin binding and IR autophosphorylation. Moreover, this mutation reduces insulin receptor substrate (IRS)-1 tyrosine phosphorylation and insulin-induced metabolic and mitogenic effects. Altogether, these observations support a role of the extracellular domain of IR beta-subunit in insulin and receptor intracellular targeting as well as in insulin signaling. DESIGN AND METHODS: This study assesses in more details the effect of IR(C860S) mutation on the trafficking of the insulin-IR complex. In particular, IR internalization, phosphorylation, dissociation and recycling, as well as insulin degradation and retroendocytosis have been investigated in CHO cells overexpressing either wild type (CHO-IR(WT)) or mutated IRs. RESULTS: the C860S mutation significantly decreases IR internalization both insulin stimulated and constitutive. In spite of a similar dissociation of internalized insulin-IR complex, recycling of internalized IR was significantly faster (half life (t(1/2)): 21 min vs 40 min, P<0.001) and more extensive (P<0.01) for IR(C860S) than for IR(WT). On the other hand, insulin degradation and retroendocytosis were superimposable in both cell lines. As expected, insulin-induced phosphorylation was similar in both IRs, however dephosphorylation was much more rapid and was greater (P<0.01) in CHO-IR(WT) as compared with CHO-IR(C860S) cells. CONCLUSIONS: Transmembrane and intracellular domain of IR seem to be determinants for IR internalization. Now we report that Cys 860 in the IR beta-subunit ectodomain may be of relevance in ensuring a proper internalization and intracellular trafficking of the insulin-IR complex.
Asunto(s)
Espacio Extracelular/metabolismo , Insulina/metabolismo , Receptor de Insulina/fisiología , Animales , Células CHO , Cricetinae , Semivida , Humanos , Fosforilación , Fosfotirosina/metabolismo , Receptor de Insulina/genética , Receptor de Insulina/metabolismo , TransfecciónRESUMEN
A(3) adenosine receptors have been proposed to play an important role in the pathophysiology of cerebral ischemia with a regimen-dependent nature of the therapeutic effects probably related to receptor desensitization and down-regulation. Here we studied the agonist-induced internalization of human A(3) adenosine receptors in transfected Chinese hamster ovary cells, and then we evaluated the relationship between internalization and signal desensitization and resensitization. Binding of N(6)-(4-amino-3-[(125)I]iodobenzyl)adenosine-5'-N-methyluronamide to membranes from Chinese hamster ovary cells stably transfected with the human A(3) adenosine receptor showed a profile typical of these receptors in other cell lines (K:(D) = 1.3+/-0.08 nM; B(max) = 400+/-28 fmol/mg of proteins). The iodinated agonist, bound at 4 degrees C to whole transfected cells, was internalized by increasing the temperature to 37 degrees C with a rate constant of 0.04+/-0.034 min(-1). Agonist-induced internalization of A(3) adenosine receptors was directly demonstrated by immunogold electron microscopy, which revealed the localization of these receptors in plasma membranes and intracellular vesicles. Moreover, short-term exposure of these cells to the agonist caused rapid desensitization as tested in adenylyl cyclase assays. Subsequent removal of the agonist led to restoration of the receptor function and recycling of the receptors to the cell surface. The rate constant of receptor recycling was 0.02+/-0.0017 min(-1). Blockade of internalization and recycling demonstrated that internalization did not affect signal desensitization, whereas recycling of internalized receptors was implicated in the signal resensitization.
Asunto(s)
Adenosina/análogos & derivados , Endocitosis/fisiología , Agonistas del Receptor Purinérgico P1 , Receptores Purinérgicos P1/metabolismo , Adenosina/farmacocinética , Marcadores de Afinidad/farmacocinética , Animales , Unión Competitiva/efectos de los fármacos , Células CHO , Membrana Celular/metabolismo , Concanavalina A/farmacología , Cricetinae , Relación Dosis-Respuesta a Droga , Endocitosis/efectos de los fármacos , Humanos , Concentración de Iones de Hidrógeno , Soluciones Hipertónicas/farmacología , Inmunohistoquímica , Radioisótopos de Yodo/análisis , Ligandos , Potasio/metabolismo , Ensayo de Unión Radioligante , Receptor de Adenosina A3 , Receptores Purinérgicos P1/genética , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Temperatura , TransfecciónRESUMEN
There is evidence that intracellular insulin may carry out some insulin mediated actions, including glucose transport. As intracellular insulin has never been quantitatively assessed in human cells, we evaluated its concentrations in monocytes from normal subjects (n = 7) and obese patients without (n = 9) and with Type 2 diabetes mellitus (n = 10). After the incubation of cells with labeled insulin for 60 min at 37 degrees C, intracellular intact insulin concentrations were measured by HPLC and expressed as pmol x 10(-6). Insulin concentrations were higher (ANOVA P < 0.01) within cells from obese (115.4 +/- 26.4 pmol x 10(-6)/2 x 10(5) cells) and obese diabetic patients (93.2 +/- 36.3 pmol x 10(-6)/2 x 10(5) cells) compared with normal cells (28.5 +/- 13.1 pmol x 10(-6)/2 x 10(5) cells). Moreover, after insulin was removed from the incubation medium the decrease of intracellular insulin was significantly lower (P < 0.01) in cells from both obese and obese diabetic patients than in normal subjects. Intracellular undissociated insulin-insulin receptor complexes on average, increased 2-fold (P < 0.01) in cells from insulin resistant patients compared with normal cells. Finally, in downregulated cells from obese and obese diabetic patients, the recycling of the internalized insulin receptor was completely disrupted. In conclusion, monocytes from obese patients with and without Type 2 diabetes mellitus, present increased intracellular insulin concentrations and these conditions are associated with a significant impairment of insulin receptor processing. Increased intracellular insulin concentration in cells from these patients may be necessary in order to overcome insulin resistance.
Asunto(s)
Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus/sangre , Hiperinsulinismo/sangre , Insulina/sangre , Monocitos/metabolismo , Obesidad/sangre , Cromatografía Líquida de Alta Presión , Humanos , Hiperinsulinismo/etiología , Membranas Intracelulares/metabolismo , Persona de Mediana Edad , Concentración Osmolar , Receptor de Insulina/metabolismoAsunto(s)
Supervivencia de Injerto , Trasplante de Islotes Pancreáticos/fisiología , Trasplante Heterólogo/fisiología , Animales , Bovinos , Células Cultivadas , Cromatografía Líquida de Alta Presión , Femenino , Insulina/sangre , Insulina/metabolismo , Secreción de Insulina , Islotes Pancreáticos/citología , Trasplante de Islotes Pancreáticos/métodos , Trasplante de Islotes Pancreáticos/patología , Porcinos , Timo , Trasplante Heterólogo/métodos , Trasplante Heterólogo/patología , Trasplante HeterotópicoRESUMEN
Leptin can be considered as a peripheral signal which informs the centers about the mass of energy stores. Studies done on the human adult population have demonstrated that degree of adiposity and insulin levels play a major role as determinants of leptin circulating levels. The aim of this study was to evaluate which factors may influence leptin levels at birth. We examined the role played by baby size and by the metabolic environment the fetus was exposed to during pregnancy. We considered 85 newborns from normal (n = 60), gestational (GDM, n = 17) and pregestational (IDDM = 8) diabetes mellitus mothers. At delivery, blood was taken from the umbilical cord vein. Babies from normal and GDM mothers were subdivided into AGA (appropriate for gestational age) and LGA (large for gestational age). There was no difference in leptin levels between babies from normal or GDM mothers belonging to the same weight category, but leptin levels were always higher in LGA than in AGA newborns, and highly correlated with birth weight (r = 0.34, P = 0.001). Moreover, IDDM mothers gave birth to newborns with significantly higher levels of leptin and insulin when compared with normal and GDM mothers. Diabetes of both GDM and IDDM mothers was clinically well controlled (HbA1c was 4.0 and 7.2, respectively). The correlation between leptin and insulin was significant only when newborns from IDDM mothers were included in the regression analysis (r = 0.39, P = 0.0002). Our results suggest that degree of adiposity is one of the main regulators of leptin concentration in the human newborn and that babies exposed to an altered, though clinically controlled, metabolic environment, as in IDDM mothers, have increased levels of leptin.
Asunto(s)
Diabetes Mellitus Tipo 1/sangre , Diabetes Gestacional/sangre , Proteínas/metabolismo , Tejido Adiposo/metabolismo , Adulto , Peso al Nacer , Glucemia , Péptido C/sangre , Femenino , Hemoglobina Glucada/análisis , Humanos , Recién Nacido , Insulina/sangre , Proteína 1 de Unión a Factor de Crecimiento Similar a la Insulina/sangre , Factor I del Crecimiento Similar a la Insulina/metabolismo , Leptina , Análisis Multivariante , Embarazo , Análisis de Regresión , Testosterona/sangreRESUMEN
Dissociation of the insulin-insulin receptor complex plays a crucial role in the processing of both insulin and the insulin receptor, and the acidification of endocytic vesicles may be the mechanism by which internalized insulin is dissociated from its receptor and properly sorted and processed. Internalized insulin-insulin receptor complexes are abnormally processed in cells from patients with non-insulin-dependent diabetes mellitus (NIDDM). Accordingly, to further investigate the mechanisms of the derangements observed in NIDDM cells, we examined the effects of the ionophore monensin, which inhibits endosomal acidification, on the cellular processing of insulin and insulin receptor in monocytes from control subjects (n = 12) and NIDDM patients (n = 14). This study confirms that monocytes from NIDDM patients, compared with cells from normal controls, had reduced binding (P < .01), internalization (P < .01), and degradation (P < .01) of insulin. In addition, the release of intracellular radioactivity was slower (P < .01), and recycling of the insulin receptor was inhibited (P < .01). Moreover, these defects were associated with a significant (P < .01) decrease of dissociation of the internalized insulin-insulin receptor complex. In cells from normal controls, incubation with monensin decreased insulin binding (P < .01), but not insulin internalization. High-performance liquid chromatography (HPLC) analysis of intracellular radioactivity showed that after monensin intracellular intact insulin significantly increased (P < .01), thus suggesting a decrease of intracellular insulin degradation. Moreover, insulin receptor recycling was completely disrupted. All of these derangements were associated with a significant decrease (P < .01) of dissociation of insulin-insulin receptor complexes. On the contrary, in diabetic monocytes, monensin had no significant additional effect on NIDDM-linked alterations. Comparison of the results obtained in cells from NIDDM patients to those found in monensin-treated normal cells demonstrates that NIDDM and monensin gave rise to a superimposable impairment of dissociation of the intracellular insulin-insulin receptor complex, associated with similar abnormal sorting and processing of insulin and its receptor. The only defect present in NIDDM cells but not in monensin-treated cells is the decrease of insulin internalization, which thus seems independent of the action of monensin on the processing of internalized insulin-insulin receptor complex. These results suggest that the impairment of dissociation of the insulin-insulin receptor complex may play a crucial role in the subsequent altered processing of insulin and insulin receptor. Moreover, they raise the question as to a possible similar alteration of the same intracellular mechanism by NIDDM and monensin, and point out that the derangements found in cells from NIDDM patients could be localized within the endosomal apparatus and consist mainly of a defective acidification of its interior.
Asunto(s)
Diabetes Mellitus Tipo 2/sangre , Endocitosis/fisiología , Endosomas/metabolismo , Insulina/análogos & derivados , Monocitos/metabolismo , Receptor de Insulina/metabolismo , Cromatografía Líquida de Alta Presión , Estudios de Cohortes , Diabetes Mellitus Tipo 2/fisiopatología , Endocitosis/efectos de los fármacos , Endosomas/efectos de los fármacos , Humanos , Insulina/metabolismo , Radioisótopos de Yodo , Ionóforos/análisis , Ionóforos/metabolismo , Ionóforos/farmacología , Persona de Mediana Edad , Monensina/análisis , Monensina/metabolismo , Monensina/farmacología , Monocitos/efectos de los fármacos , Unión Proteica/efectos de los fármacos , Unión Proteica/fisiología , Receptor de Insulina/efectos de los fármacos , Valores de Referencia , Factores de TiempoRESUMEN
Bovine islets are being evaluated for their potential in transplantation studies. We studied the recovery, morphology, and function of purified bovine islets cultured up to 4 weeks under varying conditions. Approximately 60% of the initial islet mass could be recovered after 4 weeks at 37 degrees C in CMRL 1066 or M 199 culture medium, and the cultured islets were well preserved histologically and viable both in vitro and in vivo. On the other hand, culture with RPMI 1640 caused disaggregation of the islets within a few days, with altered in vitro viability. Thus, culturing purified bovine islets with appropriate media is a suitable procedure to maintain islet mass, morphology, and function in the long term.
Asunto(s)
Medios de Cultivo/farmacología , Insulina/metabolismo , Islotes Pancreáticos/fisiología , Animales , Bovinos , Inmunohistoquímica , Secreción de Insulina , Islotes Pancreáticos/efectos de los fármacos , Ratones , Trasplante de ÓrganosRESUMEN
This study evaluates the prevalence of diabetes mellitus (DM) in Pisa (Tuscany, Italy) using four independent data sources. The main source, represented by computerized prescriptions for anti-diabetic agents collected over a 4-month period, was validated using three secondary sources: (a) the list of diabetic patients who receive material of self-care from the National Health Service; (b) the clinical records of diabetic patients obtained from a random sample of family doctors; (c) the clinical records of diabetic patients attending our outpatient clinic. The main source provided 3806 patients, and 697 patients were added from the secondary sources, thus identifying a total number of 4503. The prevalence of known DM in the "Pisa area" exclusively reckoned by the main source, was 2.01%, and the prevalence corrected by the addition of the various sources resulted in 2.4%. The capture-recapture method showed a completeness of ascertainment of the survey of 90.1%, and thus an estimated prevalence of known diabetes of 2.64%. Of these, 141 patients had insulin-dependent diabetes mellitus (IDDM) corresponding to 3.2% of identified diabetic subjects (prevalence 0.07% inhabitants); 4362 patients had non-insulin-dependent diabetes mellitus (NIDDM), 96.8% of identified diabetic subjects (prevalence 2.36%). Of patients with NIDDM 10.5% was treated by diet, 65% with oral hypoglycaemic agents (OHA), 23% with insulin and 1.5% with insulin plus OHA. This study shows that the method used in this survey is suitable for epidemiological studies because it does not demand the cooperation of the diabetic patients, is addressed to the entire diabetic population without age discrimination and singles out the diabetic population in a very reliable way.
Asunto(s)
Diabetes Mellitus/epidemiología , Prescripciones de Medicamentos/estadística & datos numéricos , Adulto , Factores de Edad , Edad de Inicio , Anciano , Instituciones de Atención Ambulatoria , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Diabetes Mellitus Tipo 1/epidemiología , Diabetes Mellitus Tipo 2/dietoterapia , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/epidemiología , Medicina Familiar y Comunitaria , Femenino , Humanos , Insulina/uso terapéutico , Italia/epidemiología , Masculino , Obesidad , Prevalencia , Factores SexualesRESUMEN
The products of insulin metabolism generated in vitro and in vivo were compared in this study. Monocytes from 10 control subjects were incubated with 125IA14-labeled insulin, acid washed, and solubilized or reincubated in insulin-free binding buffer to study both intracellular radioactivity or radioactivity released from cells to medium. To evaluate in vivo insulin metabolism, labeled insulin (100-120 microCi) was injected as a single intravenous bolus in 5 of the 10 subjects. Cellular and plasma radioactivity was characterized by high-performance liquid chromatography (HPLC). The results of the study show the following: 1) Products with superimposable HPLC elution profiles are found within cells and in medium. Two new labeled products are observed in the latter, suggesting that a membrane degradation process exists in monocytes. 2) Intermediates found within monocytes, in medium from monocytes, and in plasma have identical elution profiles, supporting the possibility that insulin is metabolized in various cells by a common pathway. 3) Insulin metabolism produces intermediates that bind well to anti-insulin antibody. The presence in plasma of these products induces a significant difference in the value of the metabolic clearance rate of insulin when HPLC or immunoprecipitation is used to detect intact insulin. 4) Immunoprecipitable products maintain, in part, the capacity to bind to insulin receptors and to be internalized into monocytes.
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Insulina/sangre , Leucocitos Mononucleares/metabolismo , Cromatografía Líquida de Alta Presión , Humanos , Técnicas In Vitro , Inyecciones Intravenosas , Insulina/administración & dosificación , Insulina/análogos & derivados , Radioisótopos de Yodo , Cinética , Masculino , Técnica de Dilución de Radioisótopos , Valores de Referencia , Factores de TiempoAsunto(s)
Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus/tratamiento farmacológico , Gliburida/uso terapéutico , Insulina Isófana/uso terapéutico , Obesidad , Fenformina/uso terapéutico , Glucemia/metabolismo , Péptido C/sangre , Diabetes Mellitus/sangre , Diabetes Mellitus/dietoterapia , Diabetes Mellitus Tipo 2/sangre , Esquema de Medicación , Quimioterapia Combinada , Glucagón , Gliburida/administración & dosificación , Hemoglobina Glucada/análisis , Humanos , Insulina Isófana/administración & dosificación , Persona de Mediana Edad , Fenformina/administración & dosificación , Factores de Tiempo , Pérdida de PesoAsunto(s)
Diabetes Mellitus Tipo 1/epidemiología , Insulina/uso terapéutico , Adolescente , Adulto , Factores de Edad , Anciano , Niño , Preescolar , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Femenino , Humanos , Lactante , Italia/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Factores SexualesRESUMEN
The authors describe the diurnal profile of plasma metformin concentrations in a group of 6 Type 2 (noninsulin-dependent) diabetic patients studied at two different daily metformin doses (500 mg and 850 mg b.d.) and report data on the relationships between plasma metformin and metabolic effects over a 14 h period. In addition, the effect of circulating metformin on insulin binding to isolated monocytes has been evaluated. At the two different daily doses fasting plasma metformin concentrations were similar (3.23 +/- 0.35 mumol/l and 3.86 +/- 0.72 mumol/l, mean values +/- SEM, at low and high dose respectively). Drug peak values and averaged concentrations (4.66 +/- 0.39 mumol/l vs 6.35 +/- 0.69 mumol/l) were significantly higher when more drug was administered. Mean plasma glucose was lower when 1,700 mg/day instead of 1,000 mg/day of metformin was given (7.3 +/- 0.4 mmol/l vs 9.1 +/- 0.9 mmol/l, p less than 0.05). After dosing, at higher plasma metformin concentrations corresponded lower plasma glucose values. The averaged blood lactate levels resulted 1.46 +/- 0.4 mmol/l (p less than 0.05 vs matched diet treated diabetic patients) at the higher drug dose. A significant positive correlation emerged between mean plasma metformin concentrations and mean blood lactate levels (r: 0.76, p less than 0.02). Alanine, glycerol and B-OH-butyrate levels were similar at the two metformin daily doses, and were not correlated to plasma metformin. The binding of insulin to isolated human monocytes was similar in metformin-treated diabetic patients (4.48 +/- 0.45) as in healthy volunteers (4.62 +/- 0.34); insulin binding was correlated (p less than 0.05) with plasma metformin levels.
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Ritmo Circadiano/fisiología , Diabetes Mellitus Tipo 2/sangre , Metformina/sangre , Alanina/sangre , Glucemia/metabolismo , Butiratos/sangre , Ácido Butírico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Glicerol/sangre , Humanos , Lactatos/sangre , Ácido Láctico , Metformina/uso terapéutico , Piruvatos/sangre , Ácido PirúvicoRESUMEN
The metabolism of insulin in vivo was investigated using an isocratic reversed-phase high-performance liquid chromatographic (RP-HPLC) method. After intravenous injection of A14-[125I]insulin into normals, eight labelled insulin derivatives were found in plasma (peaks 1-8). Two of them (peaks 1 and 7) showed an elution pattern identical with those of reference [125I]monoiodotyrosine and intact A14-[125I]insulin, respectively. Of the other six peaks, five (2-6) eluted before and one (peak 8) after insulin. This pattern was highly reproducible in terms of capacity factors and peak heights. Radioactivity separated by RP-HPLC was further characterized for its trichloroacetic acid precipitability and immunoprecipitability. Fractions corresponding to peaks 4-6 and 8, which showed an immunoprecipitability higher than 50%, were pooled in order to obtain sufficient radioactivity and were found to be insulin separated by Sephadex G-50 chromatography, containing in its structure, after sulphitolysis, intact A-chain and to be partially rebindable to monocyte insulin receptors. These data demonstrate that in blood, products of insulin metabolism circulate which retain a part of the immunological and biological properties of the hormone. These products are clearly separated from one another and from intact insulin by RP-HPLC, suggesting that the appropriate use of this technique may allow a further and more accurate qualitative and quantitative characterization of in vivo insulin metabolism in physiological and pathological conditions.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Insulina/metabolismo , Humanos , Radioisótopos de Yodo , Pruebas de Precipitina , Ácido TricloroacéticoRESUMEN
This study tests the possibility to avoid the use of phenformin in 40 type 2 (non insulin dependent) diabetic patients treated with the commercial sulphonylurea-phenformin combinations. In diabetic patients treated with sulphonylureas and phenformin at low dosage (glibenclamide 5 mg and phenformin 50 mg) it was possible to maintain good glycometabolic control using only the sulphonylurea gliclazide (160 mg/die). The diabetic patients on treatment with sulphonylureas and phenformin at higher dosage (glibenclamide 7.5 mg and phenformin 75 mg) may further improve their metabolic control when transferred to a gliclazide-benfluorex combination 160 mg and 300 mg/die, respectively. These results suggest the possibility of withdrawing or replacing phenformin in the therapy of type 2 diabetic patients without modifying their glycometabolic control.
Asunto(s)
Diabetes Mellitus Tipo 2/tratamiento farmacológico , Fenfluramina/análogos & derivados , Gliclazida/administración & dosificación , Gliburida/administración & dosificación , Fenformina/administración & dosificación , Anciano , Glucemia/análisis , Diabetes Mellitus Tipo 2/metabolismo , Quimioterapia Combinada , Femenino , Fenfluramina/administración & dosificación , Humanos , Masculino , Persona de Mediana Edad , Factores de TiempoRESUMEN
This study investigated the relative effect of obesity alone and in combination with non-insulin-dependent diabetes mellitus (NIDDM) on the intracellular processing of insulin and evaluated the effect of metformin therapy on this process. Monocytes from 11 obese hyperinsulinemic subjects, 13 obese hyperinsulinemic NIDDM patients, and 7 nondiabetic control subjects were incubated with A14-125I-labeled insulin for 60 min at 37 degrees C, and intracellular insulin degradation was characterized by high-performance liquid chromatography. Total cell-associated insulin (insulin binding) and internalized and degraded insulin were decreased in obese subjects and significantly decreased in obese NIDDM patients compared with nondiabetic control subjects. In NIDDM patients, intracellular insulin degradation was inversely correlated with fasting plasma glucose (P less than 0.01). Eight obese subjects and 9 obese NIDDM patients were restudied after 4 wk of therapy with metformin (850 mg twice a day). Plasma levels of the drug were superimposable in the two groups. Metformin therapy did not change glucose and insulin levels in obese subjects but caused a decrease in blood glucose in obese NIDDM patients. Total cell-associated radioactivity (insulin binding) significantly increased in both groups (P less than 0.01). On the contrary, internalized radioactivity increased (0.83 +/- 0.3 vs. 1.31 +/- 0.35%, P less than 0.01), and similarly, insulin degradation was enhanced (54.6 +/- 8.9 vs. 74.22 +/- 9.15%, P less than 0.01) only in monocytes from obese NIDDM patients. However, the levels of these parameters were still lower than in control subjects (internalization, 2.94 +/- 0.68%; degradation, 93.03 +/- 3.7%).(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus/sangre , Insulina/sangre , Metformina/farmacología , Monocitos/metabolismo , Transporte Biológico/efectos de los fármacos , Glucemia/metabolismo , Humanos , Cinética , Persona de Mediana Edad , Monocitos/efectos de los fármacos , Obesidad/sangre , Valores de ReferenciaRESUMEN
Salivary immunoreactive insulin was measured in 45 insulin-treated diabetic patients directly from saliva samples by a radioimmunoassay procedure. Mean and median fasting salivary immunoreactive insulin was respectively 88.5 and 54.0 pmol/L (range 12.2-633.6 pmol/L); fasting salivary insulin values were positively correlated with steady-state plasma free-insulin levels measured in immediately extracted plasma samples (r = 0.62, p less than 0.001) and with daily insulin dose (r = 0.32, p less than 0.05). After subcutaneous injection of 15 U regular insulin in 6 type 1 diabetic patients, salivary insulin concentrations increase reflected that of plasma free-insulin during a 6 h period although with a 60 min time lag. Mean salivary insulin values and plasma free-insulin concentrations were significantly correlated to each other both at each time point (p less than 0.001) and when the mean salivary insulin levels were plotted against the mean plasma free-insulin values 60 min earlier (p less than 0.001). These results suggest that the measurement of salivary immunoreactive insulin (that is easier, faster and cheaper than plasma free-insulin assay) may be usefully employed as an approximation of plasma free-insulin concentrations for clinical and research purposes in patients with circulating anti-insulin antibodies.
Asunto(s)
Diabetes Mellitus Tipo 1/metabolismo , Anticuerpos Insulínicos/análisis , Insulina/metabolismo , Saliva/análisis , Adulto , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Diabetes Mellitus Tipo 1/inmunología , Humanos , Insulina/análisis , Insulina/uso terapéutico , Cinética , Radioinmunoensayo , Factores de TiempoRESUMEN
Diabetes mellitus was assessed by investigating 8 type I diabetic patients, in good metabolic control and with congestive heart failure, who were studied after 3 weeks of placebo and ibopamine (100 mg t.i.d.) treatment. Metabolic control and daily insulin dose did not change in any patient during the study. The insulin-mediated glucose uptake during the clamp studies showed no variation after placebo or ibopamine therapy. Total cholesterol, HDL-cholesterol and triglycerides concentrations remained unchanged. This study clearly suggests that ibopamine administered at a daily dose of 300 mg for 3 weeks presents a metabolic safety in type I diabetic patients.