ABSTRACT
Pentraxin 3 (PTX3) is a tumor necrosis factor and interleukin-1beta-stimulated gene that encodes a long PTX with proinflammatory activity. Here, we show that peritoneal macrophages derived from PTX3 transgenic (Tg) mice express higher levels of PTX3 mRNA than macrophages from wild-type (WT) mice, at basal level as well as upon stimulation with zymosan (Zy). Macrophages from Tg mice also showed improved opsonin-independent phagocytosis of Zy particles and the yeast form of the fungus Paracoccidioides brasiliensis. In the case of P. brasiliensis, an enhanced microbicidal activity accompanied by higher production of nitric oxide was also observed in macrophages from Tg mice. Using fluorescein-activated cell sorter analysis and reverse transcriptase-polymerase chain reaction, we demonstrated that basal level of Toll-like receptor-6 and Zy-induced dectin-1 expression was slightly but consistently higher in macrophages from Tg mice than in macrophages from WT mice. Recombinant (r)PTX3 protein binds to Zy particles as well as to yeast cells of P. brasiliensis and addition of rPTX3, to a culture of WT-derived macrophages containing Zy leads to an increase in the phagocytic index, which parallels that of Tg-derived macrophages, demonstrating the opsonin-like activity of PTX3. It is important that blockade of dectin-1 receptor inhibited the phagocytosis of Zy particles by WT and PTX3 Tg macrophages, pointing out the relevant role of dectin-1 as the main receptor involved in Zy uptake. Our results provide evidence for a role of PTX3 as an important component of the innate-immune response and as part of the host mechanisms that control fungal recognition and phagocytosis.
Subject(s)
C-Reactive Protein/genetics , Macrophages, Peritoneal/metabolism , Membrane Proteins/metabolism , Nerve Tissue Proteins/metabolism , Opsonin Proteins/genetics , Phagocytosis/genetics , Serum Amyloid P-Component/genetics , Zymosan/immunology , Animals , Binding Sites/drug effects , Binding Sites/genetics , C-Reactive Protein/metabolism , Female , Immunity, Innate/genetics , Lectins, C-Type , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/immunology , Male , Membrane Glycoproteins/drug effects , Membrane Glycoproteins/metabolism , Membrane Proteins/antagonists & inhibitors , Mice , Mice, Transgenic , Nerve Tissue Proteins/antagonists & inhibitors , Nitric Oxide/metabolism , Opsonin Proteins/metabolism , Paracoccidioides/immunology , Phagocytosis/drug effects , Phagocytosis/immunology , RNA, Messenger/drug effects , RNA, Messenger/metabolism , Receptors, Cell Surface/drug effects , Receptors, Cell Surface/metabolism , Recombinant Fusion Proteins/metabolism , Serum Amyloid P-Component/metabolism , Toll-Like Receptor 6 , Zymosan/metabolism , Zymosan/pharmacologyABSTRACT
AIM: The goal of this study was to test the hypothesis that insulin receptor tyrosine kinase activity of isolated erythrocytes would be greater in glyburide-treated patients with type 2 diabetes in good glycaemic control (n = 13) than in untreated patients (n = 12) with significant fasting hyperglycaemia. METHODS: The two groups were similar in age, sex distribution, and body mass index. By selection, glyburide-treated patients had significantly (p < 0.001) lower (mean +/- s.e.m.) fasting glucose (6.9+/-0.4 vs. 13.9+/-0.8 mmol/l) and HbA(IC) (7.4+/-0.2 vs. 11.8+/-0.9%) concentrations. In addition, insulin-stimulated tyrosine kinase activity was increased in erythrocytes from glyburide -treated patients (p < 0.01). RESULTS: Although insulin receptor number was similar in solubilized erythrocytes from the two groups, tyrosine kinase activity per insulin receptor was significantly (p < 0.02) greater in erythrocytes from glyburide-treated patients with type 2 diabetes. CONCLUSIONS: These findings are quite similar to previously published data in metformin-treated patients. As such, it is suggested that decreases in insulin receptor tyrosine kinase activity may contribute to the loss of insulin sensitivity in hyperglycaemic subjects (glucotoxicity), and that an improvement in glycaemic control, irrespective of how it is achieved, will help rectify this abnormality.
Subject(s)
Blood Glucose/metabolism , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/drug therapy , Erythrocytes/metabolism , Glyburide/therapeutic use , Hypoglycemic Agents/therapeutic use , Receptor, Insulin/blood , Blood Glucose/drug effects , Diabetes Mellitus, Type 2/enzymology , Female , Humans , Male , Middle Aged , Receptor, Insulin/drug effectsABSTRACT
The purpose of this study was to document the possible effect of solubilised erythrocytes on insulin-receptor binding and tyrosine kinase activity after 12 weeks of metformin administration to 13 healthy obese women with no history of diabetes and normal glucose as evaluated by conventional criteria. Subjects were given metformin 850 mg twice a day for 12 weeks. The results showed that plasma glucose response to an oral glucose challenge did not change following metformin, but that insulin response was significantly lower (p < 0.0001). In addition, both the number of insulin receptors and the tyrosine kinase activity per receptor of solubilised erythrocytes were significantly greater following metformin administration. Since both body weight and plasma glucose concentrations were similar before and after treatment, the effect of metformin on insulin-receptor binding and tyrosine kinase activity appeared to be independent of either of these variables. In summary, oral administration of metformin led to an increase in tyrosine kinase activity or erythrocyte insulin receptors, suggesting that such action occurs in the absence of any significant change in plasma glucose concentration.
Subject(s)
Blood Glucose/metabolism , Erythrocyte Membrane/metabolism , Erythrocytes/metabolism , Hypoglycemic Agents/pharmacology , Insulin/pharmacology , Metformin/pharmacology , Obesity/blood , Protein-Tyrosine Kinases/blood , Receptor, Insulin/blood , Receptor, Insulin/metabolism , Adult , Blood Glucose/drug effects , Erythrocytes/drug effects , Female , Glucose Tolerance Test , Humans , Insulin/blood , Kinetics , Obesity/enzymology , Receptor, Insulin/drug effectsABSTRACT
This study was performed to define the effect of metformin on glycaemic control and erythrocyte insulin receptor tyrosine kinase activity in patients with non-insulin-dependent (Type 2) diabetes mellitus. A case-control study of the effect of metformin treatment in hyperglycaemic patients with Type 2 diabetes was conducted in outpatients of the Diabetes Clinical Center. The study population consisted of 14 patients with Type 2 diabetes (5 males, 9 females) whose hyperglycaemia was uncontrolled by diet. Patients were treated with metformin 850 mg twice daily for 2 1/2 months. Fasting plasma glucose concentrations decreased from 8.9 to 6.4 mmol/L after 10 weeks of metformin treatment (p < 0.001), in association with significantly lower (p < 0.001) plasma glucose and insulin concentrations in response to an oral glucose load. In addition, both fasting plasma triglyceride and cholesterol concentrations were significantly (p < 0.001) lower after metformin treatment. There was no change in erythrocyte insulin receptor binding associated with metformin treatment, but both basal and insulin-stimulated insulin receptor tyrosine kinase activities of solubilized erythrocyte insulin receptors were significantly higher after 10 weeks of metformin treatment. It is concluded that the increase in insulin-stimulated tyrosine kinase activity contributed to the improvement in glucose insulin and lipoprotein metabolism associated with metformin treatment of Type 2 diabetes.
