Diabetes In Experimental Animals Research Articles

The effects of diabetes with hyperlipidemia on P450 expression in APA hamster livers.

The effect of chronic hyperglycemia and hyperlipidemia induced by streptozotocin (SZ) on the expression of P450 in the liver of APA hamsters was studied in this experiment. No effect on the total activity of P450 was seen in SZ-induced diabetic hamsters throughout the experimental period. At 1 and 6 months after SZ-injection, the levels of CYP1A, 2C6, and 3A of SZ-injected hamsters were much lower than those of age-matched control hamsters. CYP2B expression tended to decrease and CYP2E1 and 4A expression tended to increase in SZ-injected hamsters, although the results were not significant. At 3 months after SZ-injection, however, no significant difference between SZ-injected and normal hamsters was seen in these P450 isozymes. On the other hand, CYP2C11 expression was slightly depressed in SZ1M and SZ6M, and almost equivalent to control hamsters in SZ3M. Immunohistochemistry by the use of each isozyme antibody revealed that SZ-induced diabetes affected the localization of CYP2C6, 3A, and 4A in the hepatic acinus. The expression of CYP2C6 and 3A was depressed mainly in the periportal region of the acinus, and CYP4A expression was induced mainly in the perivenous region by SZ-induced diabetes. On the other hand, the expression pattern of CYP1A, 2B, 2C11, and 2E1 were not affected. These results demonstrate that the effects of SZ-induced diabetes on hepatic P450 differ for each isozyme in APA hamsters and also differ from those of other experimental diabetic animals, including golden hamsters.

A new concept: the use of vanadium complexes in the treatment of diabetes mellitus.

In the 21st century, patients suffering from diabetes mellitus (DM), a lifestyle-related disease, will increase more than in the 20th century. DM is threatening because of the development of many severe secondary complications, including atherosclerosis, microangiopathy, renal dysfunction and failure, cardiac abnormalities, diabetic retinopathy, and ocular disorders. Generally, DM is classified as either insulin-dependent type 1 or noninsulin-dependent type 2 DM. Type 1 DM is treated only by daily insulin injections; type 2 DM is treated by several types of synthetic therapeutic substances together with a controlled diet and physical exercise. Even with these measures, the daily necessity for several insulin injections can be painful both physically and mentally, whereas the synthetic therapeutic substances used over the long term often have side effects. For those reasons, the creation and development of a new class of pharmaceuticals for treatment of DM in the 21st century would be extremely desirable. In the last half of the 20th century, investigations of the relationships among diseases and micronutrients, such as iron, copper, zinc, and selenium, have been numerous. Research into the development of metallopharmaceuticals involving the platinum-containing anticancer drug, cisplatin, and the gold-containing rheumatoid arthritis drug, auranofin, has also been widespread. Such important findings prompted us to develop therapeutic reagents based on a new concept to replace either insulin injections or the use of synthetic drugs. After many trials, we noticed that vanadium might be very useful in the treatment of DM. Before the discovery of insulin by Banting and Best in 1921 and its clinical trial for treating DM, the findings in 1899, in which orally administered sodium vanadate (NaVO(3)) was reported to improve human DM, gave us the idea to use vanadium to treat DM. However, it has taken a long time to obtain a scientific explanation as to why the metal ion exhibits insulin-mimetic or blood-glucose lowering effects in in vitro and in vivo experiments. After investigations from many perspectives involving biochemistry and bioinorganic chemistry, vanadyl sulfate (VOSO(4)) and its complexes with several types of ligands have been proposed as useful for treating DM in experimental diabetic animals. On the basis of a mechanistic study, this article reports on recent progress regarding the development of antidiabetic vanadyl complexes, emphasizing that the vanadyl ion and its complexes are effective not only in treating or relieving both types of DM but also in preventing the onset of DM.

Advanced glycation end products, their receptors and diabetic angiopathy

The role of chronic hyperglycemia in the development of diabetic microvascular complications and in neuropathy has been clearly established by intervention studies. However, the biochemical or cellular links between elevated blood glucose levels, and the vascular lesions remain incompletely understood. This review focuses on the consequences of hyperglycemia on the formation of advanced glycation end‐products (AGEs), and on the role of AGEs and of their specific receptors (RAGE) in the functional and anatomical alterations of the vascular wall. AGEs are formed during the Mailiard reaction by the binding of aldoses on free NH2 groups of proteins, which, after a cascade of molecular rearrangements, result in molecules of brown color and specific fluorescence. Experimental studies have indicated that the binding of AGEs to RAGE activates cells, particularly monocytes and endothelial calls. Activated endothelial cells produce cytokines, and express adhesion molecules and tissue factor. The role of AGEs in increased oxidative stress, and in the functional alterations in vascular tone control observed in diabetes, in part related to a reduction in nitric oxide, is also discussed. The microvascular retinal, glomerular and nerve lesions induced by experimental diabetes in animals are prevented by an inhibitor of AGEs formation, aminoguanidine. The administration in diabetic animals of recombinant RAGE, which hinders AGEs‐RAGE interaction, prevents hyperpermeability and vascular lesions. These data suggest a central role of AGEs and RAGE in the development of chronic complications of diabetes.

Streptozotocin Induces Lipolysis in Rat Adipocytes in Vitro.

Streptozotocin (STZ) is used to induce experimental diabetes in animals and is also applied for the treatment of patients with insulinoma. The aim of the present work was to investigate the direct effect of STZ on lipolysis in isolated rat adipocytes. After the isolation, the cells were incubated in a Krebs-Ringer buffer of pH 7.4, at the temperature 37 °C for 90 min with different concentrations of STZ: 0.5, 1 or 2 mmol/l. STZ caused a significant rise in basal values (99 %, 199 %, and 377 %, respectively) and epinephrine-stimulated (1 µmol/l) lipolysis (15 %, 24 % and 46 %, respectively). Augmentation of basal lipolysis by STZ was neither restricted by insulin (1 nmol/l) nor by H-89 (an inhibitor of protein kinase A, 50 µmol/l). These results indicate the stimulatory influence of STZ on the action of hormone-sensitive lipase in isolated cells of white adipose tissue. The obtained outcomes suggest that in studies employing STZ, it is necessary to consider its direct effect upon lipolysis in adipocytes.

Pancreatic O6-Methylguanine DNA Methyltransferase Level in Streptozotocin-Induced Diabetic Rats

O 6 -methylguanine which is formed in cellular DNA by alkylating agents is a toxic and mutagenic lesion. O 6 -methylguanine is repaired by DNA repair protein, O 6 -methylguanine DNA methyltransferase. Deficient repair of O 6 -methylguanine has been suggested to be a contributory factor in the etiology of some diseases. Streptozotocin (SZ) is an alkylating compound that is often used to induce diabetes in experimental animals. We aimed to determine O 6 -methylguanine DNA methyltransferase level in pancreas of SZ-induced diabetic rats. Serum levels of glucose and fructosamine, and pancreatic O 6 -methylguanine DNA methyltransferase level were measured in the streptozotocin-induced diabetic and control rats. Serum glucose and fructosamine levels were found to be higher in the diabetic group than those in the control group (P<0.001 and P<0.001, respectively). Pancreatic 0 6 -methylguanine DNA methyltransferase level was lower in the diabetic group (68. 06 ′12. 37 fmol/mg protein) than in the control group (89. 45 ′13. 83 fmol/mg protein), (P<0.01) and there was a negative correlation between serum fructosamine level and pancreatic O 6 -methylguanine DNA methyltransferase level (r: - 0.6, P<0.02). These data imply that 0 6 -methylguanine DNA methyltransferase depletion may be one of the mechanisms for diabetogenic action of SZ in rats.

Renal function tests on diabetes-induced and non-induced APA hamsters.

Although it has been said that Syrian hamsters of the APA strain (APA hamsters) spontaneously develop glomerulosclerosis with age, more prominent and severe glomerulosclerosis with proteinuria as well as arteriosclerosis is induced in diabetic APA hamsters. In this study, in order to supply new information on APA hamsters, tests on renal function and histology were done on non-diabetic and streptozotocin (SZ)-induced diabetic APA hamsters (APA-N and APA-D, respectively), and the data were compared with those of normal Syrian (golden) hamsters (GOL). At 4, 8, 12, 20, and 32 weeks of age, the markers indicating renal function, serum urea nitrogen and creatinine levels and the urinary total protein level were measured and thereafter histological studies were done. Although there were no remarkable differences between APA-N and GOL in serum urea nitrogen and creatinine levels, APA-N excreted more urinary total protein from the early weeks of age. In APA-D, an apparent worsening in these markers indicating renal function was detected and diabetic nephropathy in this model was confirmed also in terms of renal function. In the histological studies, the major lesion observed in APA-D was diffuse glomerulosclerosis. This may mean that renal dysfunction in APA-D was mainly caused by the glomerular change and that it is similar to other experimental diabetic animals and human diabetic patients. These data show that the diabetic APA hamster is a desirable model of human diabetic nephropathy.

Possible involvement of endothelin-1 and nitric oxide in the pathogenesis of proliferative diabetic retinopathy.

Overproduction of endothelin-1 (ET-1) and nitric oxide (NO) in the retina is demonstrated in experimental diabetic animals. To clarify the possible involvement of ET-1 and NO in the pathogenesis of diabetic retinopathy, the authors examined the vitreous levels of these principal endothelium-derived vasoactive substances in patients with proliferative diabetic retinopathy (PDR). Vitreous fluid was taken from patients with PDR (ET-1, n = 12; NO, n = 12) and from patients with macular holes as controls (ET-1, n = 10; NO, n = 10) at vitreous surgery. Endothelin-1 and NO metabolites were measured by radioimmunoassay and high-performance liquid chromatography based on the Griess method, respectively. Endothelin-1 levels (mean +/- SE) were 21.5 +/- 1.7 pg/mL in the vitreous of patients with PDR and 16.7 +/- 0.7 pg/mL in the vitreous of patients with macular hole. There was a significant difference between patients with PDR and controls (P = 0.009, Mann-Whitney). Nitrate (NO3) was 49.8 +/- 5.0 micromol/L in patients with PDR and 24.2 +/- 2.8 micromol/L in patients with macula hole; it was also significantly elevated in patients with PDR (P = 0.004, Mann-Whitney), whereas nitrite (NO2) was not detected in this study. These results indicate that ET-1 and NO may be related in the pathogenesis of PDR.

The mechanism of alloxan and streptozotocin action in B cells of the rat pancreas

Alloxan and streptozotocin are widely used to induce experimental diabetes in animals. The mechanism of their action in B cells of the pancreas has been intensively investigated and now is quite well understood. The cytotoxic action of both these diabetogenic agents is mediated by reactive oxygen species, however, the source of their generation is different in the case of alloxan and streptozotocin. Alloxan and the product of its reduction, dialuric acid, establish a redox cycle with the formation of superoxide radicals. These radicals undergo dismutation to hydrogen peroxide. Thereafter highly reactive hydroxyl radicals are formed by the Fenton reaction. The action of reactive oxygen species with a simultaneous massive increase in cytosolic calcium concentration causes rapid destruction of B cells. Streptozotocin enters the B cell via a glucose transporter (GLUT2) and causes alkylation of DNA. DNA damage induces activation of poly ADP-ribosylation, a process that is more important for the diabetogenicity of streptozotocin than DNA damage itself. Poly ADP-ribosylation leads to depletion of cellular NAD+ and ATP. Enhanced ATP dephosphorylation after streptozotocin treatment supplies a substrate for xanthine oxidase resulting in the formation of superoxide radicals. Consequently, hydrogen peroxide and hydroxyl radicals are also generated. Furthermore, streptozotocin liberates toxic amounts of nitric oxide that inhibits aconitase activity and participates in DNA damage. As a result of the streptozotocin action, B cells undergo the destruction by necrosis.

Intrauterine Growth Retardation in Experimental Diabetes: Possible Role of the Placenta

Fetal growth disorders are common in pregnancy complicated by diabetes. Whereas macrosomia often occurs in infants of diabetic women, growth retardation is almost a rule in spontaneous and experimental diabetes in animals. However, it is not clear when during development growth inhibition starts and how placental pathology might affect fetal growth in maternal diabetes. In this study pregnant Wistar rats were injected (ip) with a single dose of 50 mg/kg of streptozotocin (STZ) on gestation day (GD) 2 and a blood glucose level of 200 mg/dl or more determined 24 hrs later indicated diabetes. The controls were non-treated, buffer treated or, following confirmation of diabetes, injected with a single dose of 2–6 IU of insulin (Novo Ultralente) once daily. Fetuses and placentae were collected from GD 14–20. Intrauterine growth retardation (IUGR) in STZ group was significant as early as GD 15 and persisted to GD 20. Insulin produced a significant recovery in fetal weight gain. The placentas of STZ-treated group were significantly heavier than those of the control groups. The reduction in cord length of the STZ group became apparent on GD 16 and remained so to term. The placenta of GD 14 STZ group had a thicker decidua basalis and dilated maternal sinusoids. By GD 16, the decidua basalis contained glycogen-containing decidual cells and scattered glycogen cells confirmed by Best' carmine with or without diastase. The glycogen cells of the basal zone were more abundant, and had degenerated in some sites leaving behind cysts with eosinophilic mass. The giant cells had proliferated enormously. The labyrinthine zone appeared spongy with persistent fetal mesenchyme, peri-vascular fibrosis, and enhanced placental barrier. The trophoblasts of the labyrinths also contained traces of glycogen unlike the controls. By GD 18, the decidua basalis of the STZ group was thinner than that of the controls and contained necrotic giant cells and lymphocytic aggregations. In the basal zone, the giant cells had proliferated further; more glycogen cells had degenerated. Perivascular fibrosis was still extensive in the labyrinthine zone. Bloodless maternal sinusoids, extensive vacuolization, degeneration of glycogen islands and formation of cysts characterized the labyrinthine zone. These changes varied in intensity from one area to another in the same placenta and between placentas of the same and of different litters. The development of the upper and lower jaws, elevation and fusion of palatal shelves, reduction of physiological umbilical hernia, descent of the testes, fusion of the urethral folds and separation of digits of the paws were significantly delayed in the STZ group. The consistent association of placental pathology with fetal growth retardation is suggestive of an alteration in placental function possibly contributing to IUGR in STZ-induced diabetes in rats.

Medicinal plants for treatment of diabetes mellitus.

Many plants have been used for the treatment of diabetes mellitus in Indian system of medicine and in other ancient systems of the world. Out of these only a few have been evaluated as per modern system of medicine. From many such plants only extracts have been prepared and their usefulness evaluated in experimental diabetes in animals. In some plants likeAllium cepa, Allium sativum, Ficus bengalensis, Gymnema sylvestre, Pterocarpus marsupium etc. active hypoglycemic principles have been isolated and their mechanism of action studied. Most of them seem to act directly on pancreas (pancreatic effect) and stimulate insulin level in blood. Some have extra pancreatic effect also by acting directly on tissues like liver, muscle etc. and alter favourably the activities of the regulatory enzymes of glycolysis, gluconeogenesis and other pathways. Since the plant products have less side effects, they have the potential as good hypoglycemic drugs. They may also provide clues for the development of new and better oral drugs for diabetes.

Modulation of cytochrome P450-dependent monooxygenases in streptozotocin-induced diabetic hamster: II. Reverse role of insulin in P450 activity and defluorination.

Metabolic activities of cytochrome (cyt) P450-dependent monooxygenase could be modulated by diabetic state in experimental diabetic animals. The purpose of this study is to validate the effect of insulin on the modulation of the metabolic activity of cyt P450 and the defluorination ability to inhalational anesthetics in diabetic animals. Diabetic state in golden Syrian hamsters was achieved by intraperitoneal injection of streptozotocin 40 mg/kg once a day for 4 days. After stabilization of diabetic state for 6 weeks, a regimen of insulin treatment given subcutaneously was carried out. Metabolic activities of cyt P450 were assessed by the reaction with benzo(a) pyrene, pentoxyresorufin, aniline and erythromycin (specific substrates). The metabolic activities of cyt 1A1, 2B1, 2E1 and 3A4 respectively in a NADPH-generating system in microsomal preparations of the diabetic hamsters were observed before and after insulin treatment, and were compared with the control group. The ability of defluorination was evaluated by measuring the free fluoride metabolites after incubating the microsomes with enflurane in diabetic and insulin-treated hamsters. Contents of cyt P450 isozymes were measured by electrophoresis and immunoblotting before and after insulin treatment. Pathological features of hepatocytes in diabetic hamsters were evaluated microscopically before and after insulin treatment. The defluorination of enflurane and activity of aniline hydroxylase (cyt 2E1) were successfully induced by diabetic state (P < 0.01). The pentoxyresorufin O-dealkylase (cyt 2B1) was inhibited nearly 50% in the diabetic hamster liver when compared with that of control (P < 0.01). While the activities of benzo(a)pyrene hydroxylase (cyt 1A1) and the erythromycin N-demethylase (cyt 3A4) were basically unaffected by diabetes, alterations in content of cyt P450 were parallel to the alterations in enzyme activities. Microscopically, diabetes induced vacuolization with fatty droplets in the hepatocytes. After treatment with insulin injection, the enzyme activities, protein content and pathologic features returned to the baseline similar to the control. Our data demonstrated that under diabetic state, metabolic activities of cyt P450 and its extent of defluorination would be polymorphically modulated. After administration of insulin, the activities of cyt P450 and defluorination of enflurane returned to baseline as the blood sugar level had been normalized. This could remind the clinicians of the importance of insulin treatment in the potential drug-to-drug interactions in the diabetic patients.

Poly(ADP-ribose) polymerase gene disruption conferred mice resistant to streptozotocin-induced diabetes.

Streptozotocin (STZ), a glucose analogue known to induce diabetes in experimental animals, causes DNA strand breaks and subsequent activation of poly(ADPribose) polymerase (Parp). Because Parp uses NAD as a substrate, extensive DNA damage will result in reduction of cellular NAD level. In fact, STZ induces NAD depletion and cell death in isolated pancreatic islets in vitro. Activation of Parp therefore is thought to play an important role in STZ-induced diabetes. In the present study, we established Parp-deficient (Parp-/-) mice by disrupting Parp exon 1 by using the homologous recombination technique. These mice were used to examine the possible involvement of Parp in STZ-induced beta-cell damage in vivo. The wild-type (Parp+/+) mice showed significant increases in blood glucose concentration from 129 mg/dl to 218, 370, 477, and 452 mg/dl on experimental days 1, 7, 21, and 60, respectively, after a single injection of 180 mg STZ/kg body weight. In contrast, the concentration of blood glucose in Parp-/- mice remained normal up to day 7, slightly increased on day 21, but returned to normal levels on day 60. STZ injection caused extensive necrosis in the islets of Parp+/+ mice on day 1, with subsequent progressive islet atrophy and loss of functional beta cells from day 7. In contrast, the extent of islet beta-cell death and dysfunction was markedly less in Parp-/- mice. Our findings clearly implicate Parp activation in islet beta-cell damage and glucose intolerance induced by STZ in vivo.

Insulin-mimetic vanadyl—dithiocarbamate complexes

As candidates for insulin mimetics, we prepared five vanadyl—dithiocarbamate complexes with VO(S 4) coordination mode, and determined their structures by elemental analysis, visible absorption, IR and electron spin resonance spectra; the five complexes were bis( N,N-dimethyldithiocarbamate)oxovanadium(IV) (VO-DMD), bis( N,N-diethyldithiocarbamate)oxovanadium(IV) (VO-DED), bis(pyrrolidine- N-dithiocarbamate)oxovanadium(IV) (VO-PYD), bis( N-methyl, N′- d-glucamine-dithiocarbamate)oxovanadium(IV) (VO-MGD) and bis(sarcosine- N-dithiocarbamate)oxovanadium(IV) (VO-SAD). The insulin-mimetic activities were evaluated by in vitro and in vivo experiments. These complexes inhibited the release of free fatty acid (FFA) from isolated rat adipocytes, similar to the action of insulin. Among them, VO-PYD and VO-SAD complexes were found to be the most effective. In addition, the VO-PYD complex promoted the incorporation of glucose in rat L6 muscle cells. Based on these in vitro observations, both VO-PYD and VO-SAD complexes were given to streptozotocin-induced diabetic rats (STZ-rats) intraperitoneally or orally. Serum glucose levels of STZ-rats dropped from hyperglycemic levels to the normal range within 1 or 2 days after both intraperitoneal and oral administrations of the complexes. To understand the insulin-mimetic action of the VO-PYD complex, the organ distribution of vanadium was investigated. In normal rats treated with VO-PYD complex, vanadium was distributed in almost all tissues, indicating that the action of vanadium is not peripheral. In addition, vanadium was found in bone and kidney when VO-PYD was given. On the basis of these results, the VO-PYD complex is indicated to be a good agent to treat insulin-dependent diabetes in experimental animals.

Metal-mediated DNA damage induced by diabetogenic alloxan in the presence of NADH

Alloxan is known to induce diabetes in experimental animals through destruction of insulin-producing β-cells of pancreas. The mechanism of DNA damage induced by alloxan was investigated using 32P-labeled human DNA fragments. Cu(II)-dependent DNA damage increased with the concentration of alloxan and NADH. Alloxan induced DNA cleavage frequently at thymine and cytosine residues in the presence of NADH and Cu(II). Catalase and bathocuproine, a Cu(I)-specific chelator, almost completely inhibited DNA damage, suggesting the involvement of H 2O 2 and Cu(I). Alloxan induced Cu(II)-dependent production of 8-oxodG in calf thymus DNA in the presence of NADH. UV-visible and electron spin resonance (ESR) spectroscopic studies showed that superoxide anion radical and alloxan radical were generated by the reduction of alloxan by NADH, and also by the autoxidation of dialuric acid, the reduced form of alloxan. These results suggest that the copper-oxygen complex derived from the reaction of H 2O 2 with Cu(I) participates in Cu(II)-dependent DNA damage by alloxan plus NADH and dialuric acid. The mechanism of DNA damage is discussed in relation to diabetogenic action of alloxan.

Review of alterations in endothelial nitric oxide production in diabetes: protective role of arginine on endothelial dysfunction.

Nitric oxide release from the endothelium plays an important role in regulation of vascular tone, inhibition of both platelet and leukocyte aggregation and adhesion, and inhibition of cell proliferation. These properties suggest that the level of NO production by the endothelium may play a pivotal role in the regulation of vascular disease. Analysis using mass spectrometry has revealed that NO is produced by NOS from the terminal guanidino nitrogen of the precursor amino acid L-arginine. Thus, utilization of L-arginine and conversion to NO may establish a regulatory site in the development of endothelial dysfunction. Endothelial dysfunction is characterized by defective endothelium-dependent relaxation, and some reviews regarding endothelial dysfunction in diabetes have been published. These reviews have focused on factors that might contribute to defective relaxation, some of which will not be addressed in detail in this review. The purpose of the present review is to summarize evidence that specifically supports either decreased NO production by diabetic vascular endothelium and/or impaired NO-mediated endothelium-dependent relaxation. Second, this review provides reasonable alternatives to explain some of the controversies in this research area. Third, since there is growing evidence that arginine appears to have some benefits for diabetes-associated abnormalities, this review summarizes the current state of knowledge of effects of acute and chronic administration of L-arginine on diabetesinduced endothelial dysfunction and discusses potential NOdependent and -independent mechanisms whereby therapeutic intervention with L-arginine might benefit the diabetic endothelium.

Generation of Nitric Oxide from Streptozotocin (STZ) in the Presence of Copper(II) plus Ascorbate: Implication for the Development of STZ-Induced Diabetes

Streptozotocin (STZ) is widely used as a strong inducer of insulin-dependent diabetes in experimental animals. Although nitric oxide (NO) generation from STZ has been proposed to be responsible for the toxicity of pancreatic B-cells, the mechanism is yet unknown. We found that STZ generates NO in the presence of Cu(II) plus ascorbate. In addition, nicotinamide, which is an antidiabetic agent against STZ, has been found to inhibit NO generation from STZ during the reaction with Cu(II) plus ascorbate. Since rat pancreatic islets contain both ascorbate and Cu at the concentrations of 3.5 mM and 1.0 nmol/mg protein, respectively, our present results indicate that (1>) NO generation is responsible for the development of STZ-induced diabetes and (2) mechanism for the protection of diabetes by nicotinamide is due to the inhibition of NO generation from STZ through complex formation between nicotinamide and Cu (I), which is reduced by ascorbate.

Effects of diabetes and food deprivation on shivering activity during progressive hypothermia in the rat

Diabetes mellitus and food deprivation are two conditions known to significantly reduce the ability to generate body heat during periods of acute cold stress. The purpose of this study was to determine if shivering is attenuated in the urethane-anesthetized (1.5 g/kg; i.p.), streptozotocin-induced diabetic (STZ; n = 10) and food-deprived (12-hour nocturnal fast) rat (FD; n = 11) as colonic temperature (T col) declined from baseline (36°C) to 28°C. Shivering was assessed using the mean rectified electromyographic (EMG) signal obtained from indwelling bipolar electrodes placed in the gluteus superficialis muscle. Although the mean rectified EMG progressively increased ( P ≤ 0.05) between T col of 33°C to 28°C and achieved peak activity (7.89 ± 1.80 μV) at 29°C in non-diabetic rats, shivering activity was virtually absent in the STZ group throughout cooling ( e.g. peak EMG = 0.49 ± 0.09 μV). The lack of shivering activity in STZ could partially explain the shorter time to reach 28°C (STZ, 48.5 ± 1.5 vs CON, 136.5 ± 23.0 min; P ≤ 0.05) and the divergent trends in oxygen consumption (ΔVO 2) between STZ and non-diabetic rats. In the FD group, the mean peak rectified EMG activity (3.09 ± 1.35 μV) was significantly lower ( P ≤ 0.05) than the fed group. The peak ΔVO 2 from baseline (FD, +2.11 ± 0.36 vs. CON, +4.51 ± 0.50 mlO 2/min) and the time taken to reach 28°C (FD, 73.4 ± 4.2 vs CON, 136.5 ± 23.0 min) were statistically different ( P ≤ 0.05) between groups. The results indicate that: 1) shivering thermogenesis is severely depressed and hypothermia accelerated in experimental diabetic animals as evidenced by the attenuation in mean rectified EMG and ΔVO 2, and 2) FD rats experienced a faster decline in colonic temperature than the fed group due, in part, to the relatively greater decline in shivering activity and oxygen consumption.

Brazilin stimulates the glucose transport in 3T3-L1 cells.

Brazilin (7,11b-dihydrobenz[b]indeno-[1,2-d]pyran-3,6a,9,10(6H)- tetrol) was found to have hypoglycemic action and increase glucose metabolism in experimental diabetic animals. In order to investigate the mechanism of hypoglycemic action of brazilin, the effects of brazilin on glucose transport, insulin receptor autophosphorylation, and protein kinase C(PKC) activity in 3T3-L1 cells were studied. Brazilin increased basal glucose transport in 3T3-L1 fibroblasts and adipocytes. However, insulin-stimulated glucose transport was not influenced. Autophosphorylation of the partially purified insulin receptor was not affected by brazilin treatment in 3T3-L1 fibroblasts. However, brazilin decreased the PKC activity in 3T3-L1 fibroblasts and adipocytes.

Therapeutic effect of berberine on 60 patients with non-insulin dependent diabetes mellitus and experimental research

The effects of berberine on 60 cases with non-insulin dependent diabetes mellitus and experimental research results were observed in this study. The results suggest berberine has significant effects on non-insulin dependent diabetes mellitus patients and experimental diabetes in animals in the reduction of blood glucose levels. The clinical symptoms basically disappeared and the level of serum insulin rose. The total effective rate was up to 90 percent and there were no significant side-effects. It was found that berberine has an effect on the recovery of pancreas islet β-cells, through pathological examination on the animal subjects.

Involvement of Elevated Intracellular and Extracellular ATP in the Regulation of Insulin Secretion: Therapeutic Targets in Non-Insulin-Dependent Diabetes Mellitus.

Non-insulin-dependent diabetes mellitus (NIDDM) is a heterogeneous disease resulting primarily from a variety of pancreatic beta-cell disorders and insulin resistance. Whereas insulin resistance, which constitutes a defect in insulin action, increases the risk of developing NIDDM and, as such, is a predictor of the onset of this disease, it is mostly the beta-cell dysfunction in regulating insulin secretion which yields the chronic hyperglycemia with all its associated clinical complications. The individual steps in the secretory pathway of insulin which is induced primarily by blood plasma glucose have now been identified. The transport of the sugar into the beta-cell is followed by its phosphorylation as the rate-determining step. The glycolytic metabolism of glucose-6-phosphate leads to the generation of ATP resulting in increases in beta-cell ATP pools (steady-state-levels) as well as ATP/ADP ratios, which, in turn, produce the closure of ATP-sensitive K(+) channels, thus depolarizing the beta-cell membrane and opening of Ca(2+) channels. The resulting influx of extracellular Ca(2+) and the increase in recruitment of Ca(2+) from intracellular stores in response to extracellular signals yield an increase in total [Ca(2+)](i) which activates the granular insulin secretory machinery. The intracellular beta-cell ATP pools have a key role in transducing the signals of the stimulus-secretion coupling pathway and toxins such as alloxan and streptozotocin which produce experimental diabetes in animals act by damaging mitochondrial oxidative phosphorylation, leading to permanent decreases in cellular ATP pools which, due to the sensitivity of beta-cell function to these pools, manifest itself as a form of diabetes. In addition to the major effects of blood plasma glucose in the regulation of insulin secretion, a variety of hormonal and neural factors producing endocrine and paracrine effects modulate and fine-tune beta-cell insulin secretion. The enteroinsular axis provides a linkage between the gastrointestinal tract and pancreatic beta-cells stimulus-secretion pathway. Although a powerful effect of ATP on insulin secretion was demonstrated more than 30 years ago, only recently has it been shown that beta-cells possess P(2)-purinoceptors. Extracellular ATP and its synthetic agonists are insulin secretagogues by virtue of their activation of membrane purinergic receptors which is coupled to increases in extracellular Ca(2+) influx and mobilization of Ca(2+) from internal stores resulting in insulin release from beta-cell granules. The physiological significance of extracellular ATP regulation of insulin secretion as well as the physiological source of these ATP pools have not yet been established. It has been recently demonstrated that the administration of adenine nucleotides in vivo can yield significant increases in tissue, blood (red blood cell), and blood plasma ATP pools. Increasing pancreatic beta-cell intracellular and blood plasma (extracellular) pools of ATP is a new therapeutic modality in non-insulin-dependent diabetes mellitus.