Brain Of Streptozotocin-induced Diabetic Rats Research Articles

Effect of alkaloid extract from Andrographis paniculata (Burm. f.) Nees and Phyllanthus amarus Schumach. & Thonn. on cognitive related biochemicals in the brain of streptozotocin-induced diabetic rats

IntroductionDiabetes mellitus (DM) is a chronic metabolic disorder linked to cognitive impairment and memory deficit, known as diabetes encephalopathy. This study examined the anti-diabetic and neuroprotective effects of alkaloid extracts from Andrographis paniculata (A. paniculata) and Phyllanthus amarus (P. amarus) leaves, a Traditional Chinese Medicine, on carbohydrate-metabolizing (α-amylase and α-glucosidase) enzyme, cholinesterase and purinergic enzyme activities, and antioxidants in the brains of streptozotocin (STZ)-induced diabetic rats. MethodsThe alkaloid extraction of A. paniculata and P. amarus was done. Male Wistar rats were divided into seven groups (n = 5) and were made diabetic by intraperitoneal injection of 50 mg/kg of STZ except Group I, which received saline (normal control). Group II-untreated diabetic rats (diabetic control), Group III-diabetic rats received 5 mg/kg Glibenclamide (standard drug); Group IV and V received 5 and 50 mg/kg P. amarus respectively; and Group VI and VII received 5 and 50 mg/kg A. paniculata, respectively. ResultsThe results showed that the elevated blood glucose and redox makers levels and enzyme activities were significantly increased in the untreated diabetic rats. Reduced antioxidant biomolecules (enzymatic and non-enzymatic antioxidant) levels were also observed. However, these were considerably ameliorated upon treatment with the alkaloid extracts in a dose-dependent manner. ConclusionThe study found that the alkaloid extract of P. amarus and A. paniculata leaves can modulate cognitive functioning and antioxidant profile in STZ-induced diabetic rats, suggesting it could be a promising alternative for managing diabetes and its complications, including cognitive impairment, in the treatment of diabetes and diabetes encephalopathy.

Neuroprotective and Antioxidative Potentials of Aqueous Crude Extract of Sterculia Tragacantha Leaf in Streptozotocin-Induced Diabetes in Rats

Background: In the western part of Nigeria, different medicinal plants including Sterculia tragacantha has been proved in managing diabetes mellitus with no scientific basis. This research focus on the neuroprotective and antioxidative potential of aqueous crude extract from S. tragacantha leaf (AESTL) using streptozotocin-induced diabetes using albino rat model. Methodology: AESTL was prepared (w/v) and its effect on biomarkers of inflammation; together with some oxidative stress, markers on the brain of streptozotocin-induced diabetic rats were evaluated. Results: The results showed that diabetic rats treated with AESTL exhibited an inhibitory effect on the neurotransmitters: acetylcholinesterase (AchE), butyrylcholinesterase (BChE), dopamine, serotonin, NO and Na+K+ ATPase, in a dose dependent manner, on the brain of the animals when compared with diabetic control. Furthermore, the results revealed that no similar effect was observed in the activities of catalase (CAT) and superoxide dismutase (SOD) in the brain of the control and diabetic rats treated with (150 and 300 mg/kg of AESTL). Also, this was observed on metformin (standard drug) group with similar effect with the control group but the vehicle (untreated diabetic group) when compared with the control group and diabetic rats treated with (150 and 300 mg/kg of AESTL) in CAT and SOD showed significant (p < 0.05) increase within the stipulated days of the experiments. In addition, brain content of the glutathione (GSH), glutathione-S-transferase (GST) and glutathione peroxidase (GPx) were significantly increased with co-administration of the AESTL doses when compared with the vehicle. Conclusion: Inhibitory effect of AChE, BChE and some other biomarkers of inflammation and antioxidative potentials by AESTL could be the major breakthrough in the management of diabetes mellitus.

Neuroprotective effects of mango cv. 'Ataulfo' peel and pulp against oxidative stress in streptozotocin-induced diabetic rats.

Oxidative stress has been implicated in the pathogenesis and progression of diabetes mellitus. Both can damage the brain. Mango and its by-products are sources of bioactive compounds with antioxidant properties. We hypothesized that mango cv. 'Ataulfo' peel and pulp mitigate oxidative stress in the brain of streptozotocin-induced diabetic rats. Twenty-four male Wistar rats were divided into four groups: control, untreated diabetic (UD), diabetic treated with a mango-supplemented diet (MTD), and diabetic pretreated with a mango-supplemented diet (MPD). The rats were fed the different diets for 4 weeks after diabetes induction (MTD), or 2 weeks before and 4 weeks after induction (MPD). After the intervention, serum and brain (cerebellum and cortex) were collected to evaluate gene expression, enzyme activity, and redox biomarkers. Superoxide dismutase 2 (SOD2) expression increased in the cortex of the MTD group, whereas glutathione-S-transferase p1 (GSTp1) expression was higher in the cortex of the MTD group, and cortex and cerebellum of the MPD group. SOD1 activity was higher in the cerebellum and cortex of all diabetic groups, whereas GST activity increased in the cerebellum and cortex of the MPD group. Lipid peroxidation increased in the cerebellum and cortex of the UD group; however, a mango-supplemented diet prevented this increase in both regions, while also mitigating polyphagia and weight loss, and maintaining stable glycemia in diabetic rats. We propose that mango exerts potent neuroprotective properties against diabetes-induced oxidative stress. It can be an alternative to prevent and treat biochemical alterations caused by diabetes. © 2020 Society of Chemical Industry.

The protective effect of Azadirachta excelsa leaves extract and quercetin treatment on the learning and memory impairments in relation with insulin and amylin levels in the brain of streptozotocin-induced diabetic rats

Azadirachta excelsa increases the secretion of plasma insulin with subsequent reduction of fasting blood glucose. However, its protective effect towards brain amylin and insulin levels as well as the memory and learning functions remain elusive. Thus, this study aimed to ascertain the effect of A. excelsa and quercetin consumptions on the brain amylin and insulin concentrations as well as memory and learning abilities of the streptozotocin (STZ)-induced diabetic rats. Diabetic condition was induced in the male Sprague Dawley rats by the intraperitoneal injection of 60 mg/kg body weight (bwt) STZ. The experimental animals were divided into normal control (saline), diabetic control (saline), Metformin-treated diabetic (1000 mg/kg bwt) (positive control), A. excelsa-treated diabetic (250 mg/kg bwt) and Quercetin-treated diabetic (40 mg/kg bwt) rats. All treatments were administered orally by oral gavage once daily throughout the 8 weeks of the treatment period. Amylin and insulin concentrations were determined by using the commercial rat amylin and insulin immunoassay kits, while learning and memory consolidation parameters were conducted using Morris Water Maze (MWM) paradigm. Treatment with A. excelsa had significantly (p < 0.05) improved the concentration of both brain amylin and insulin. In MWM test, the administration of A. excelsa and quercetin significantly (p < 0.05) attenuated the learning and memory impairments in STZ-induced diabetic rats. These analytical results provide evidence that A. excelsa can improve learning and memory impairments due to diabetes by increasing the amylin and insulin levels. These also suggest that, quercetin might only improve the learning and memory impairment, but not be the one that contributed to the improvement in the brain amylin and insulin concentrations.

Influence of quercetin, naringenin and berberine on glucose transporters and insulin signalling molecules in brain of streptozotocin-induced diabetic rats

Quercetin, naringenin, and berberine are plant bioactives that can cross the blood-brain barrier and offer neuroprotection. In the present study, we looked into the effect of them on expression of various glucose transporters and key components of brain insulin signalling, namely, insulin receptor substrate 1 (IRS 1), phosphatidyl inositol 3 kinase (PI3K), Akt 1 and low-density lipoprotein receptor-related protein 1 (LRP1) in brain of control, diabetic and bioactive-treated rats by Western blot. Amongst the bioactives tested, quercetin was more potent and restored LRP1 and brain insulin signalling components as well as glucose transporters such as GLUTs 1, 2, 3 and 4 in diabetic animals. On the other hand, berberine and naringenin supplementation to diabetic animals improved brain IRS 1 levels and restored GLUT 1 and GLUT 3 expression without significant effect on PI3K and Akt 1 activation and GLUT 4 levels. From the present study, we conclude that quercetin, naringenin, and berberine can differentially act through insulin-dependent and -independent mechanisms thereby altering glucose homeostasis in the brain during experimental diabetes and bring about the beneficial effect.

Neuroprotective effects of quercetin on memory and anxiogenic-like behavior in diabetic rats: Role of ectonucleotidases and acetylcholinesterase activities.

The present study investigated the protective effect of quercetin (Querc) on memory, anxiety-like behavior and impairment of ectonucleotidases and acetylcholinesterase (AChE) activities in brain of streptozotocin-induced diabetic rats (STZ-diabetes). The type 1 diabetes mellitus was induced by an intraperitoneal injection of 70mg/kg of streptozotocin (STZ), diluted in 0.1M sodium-citrate buffer (pH 4.5). Querc was dissolved in 25% ethanol and administered by gavage at the doses of 5, 25 and 50mg/kg once a day during 40days. The animals were distributed in eight groups of ten animals as follows: vehicle, Querc 5mg/kg, Querc 25mg/kg, Querc 50mg/kg, diabetes, diabetes plus Querc 5mg/kg, diabetes plus Querc 25mg/kg and diabetes plus Querc 50mg/kg. Querc was able to prevent the impairment of memory and the anxiogenic-like behavior induced by STZ-diabetes. In addition, Querc prevents the decrease in the NTPDase and increase in the adenosine deaminase (ADA) activities in SN from cerebral cortex of STZ-diabetes. STZ-diabetes increased the AChE activity in SN from cerebral cortex and hippocampus. Querc 50mg/kg was more effective to prevent the increase in AChE activity in the brain of STZ-diabetes. Querc also prevented an increase in the malondialdehyde levels in all the brain structures. In conclusion, the present findings showed that Querc could prevent the impairment of the enzymes that regulate the purinergic and cholinergic extracellular signaling and improve the memory and anxiety-like behavior induced by STZ-diabetes.

Resveratrol regulates oxidative biomarkers and antioxidant enzymes in the brain of streptozotocin-induced diabetic rats

Context: Oxidative stress has been implicated in the progression of pathogenesis in diabetes mellitus and leads to a variety of deformations in the central nervous system. Recent studies have provided several insights on therapeutic uses of resveratrol in diabetic complications.Objective: The present study determines if resveratrol ameliorates oxidative stress and molecular changes in the brain frontal cortex of streptozotocin-induced diabetic rats.Materials and methods: Rats were divided into four groups: control, diabetic, resveratrol-treated control, and resveratrol-treated diabetic. After diabetes induction, resveratrol (20 mg/kg) was given intraperitoneally once daily for 4 weeks. In addition to enzymatic activities, gene and protein expression of brain antioxidant enzymes were utilized by qRT-PCR and Western blot, respectively.Results: The results indicated a significant elevation in total oxidant species (1.22-fold) and malonedialdehyde (1.38-fold) contents in diabetic rat brain cortex tissues. In addition, significant augmentation in the activities of catalase (1.38-fold) and superoxide dismutase (3-fold) was witnessed with the gene and protein expression levels reflecting a transcriptional regulation. Resveratrol treatment significantly normalized diabetic malonedialdehyde and oxidized glutathione levels and strengthens the action of all antioxidant enzymes. Recovery of the diabetes-associated changes reflects the reduction of oxidative conditions by resveratrol and reveals the decrease in the requirement for the activation of antioxidant defense systems in the brain tissues of diabetic rats.Discussion and conclusion: Potent antioxidant and neuroprotective properties of resveratrol against diabetes-induced oxidative damage were demonstrated and the results support the conduct of new studies searching for the molecular mechanism of diabetes-induced changes in brain tissues.

Neuroprotective effect of ginger in the brain of streptozotocin-induced diabetic rats

Diabetes mellitus results in neuronal damage caused by increased intracellular glucose leading to oxidative stress. Recent evidence revealed the potential of ginger for reducing diabetes-induced oxidative stress markers. The aim of this study is to investigate, for the first time, whether the antioxidant properties of ginger has beneficial effects on the structural brain damage associated with diabetes. We investigated the observable neurodegenerative changes in the frontal cortex, dentate gyrus, and cerebellum after 4, 6, and 8 weeks of streptozotocin (STZ)-induced diabetes in rats and the effect(s) of ginger (500 mg/kg/day). Sections of frontal cortex, dentate gyrus, and cerebellum were stained with hematoxylin and eosin and examined using light microscopy. In addition, quantitative immunohistochemical assessments of the expression of inducible NO synthase (iNOS), tumor necrosis factor (TNF)-α, caspase-3, glial fibrillary acidic protein (GFAP), acetylcholinesterase (AChE), and Ki67 were performed. Our results revealed a protective role of ginger on the diabetic brain via reducing oxidative stress, apoptosis, and inflammation. In addition, this study revealed that the beneficial effect of ginger was also mediated by modulating the astroglial response to the injury, reducing AChE expression, and improving neurogenesis. These results represent a new insight into the beneficial effects of ginger on the structural alterations of diabetic brain and suggest that ginger might be a potential therapeutic strategy for the treatment of diabetic-induced damage in brain.

Flavonoid, morin inhibits oxidative stress, inflammation and enhances neurotrophic support in the brain of streptozotocin-induced diabetic rats

Diabetes-induced damages in brain are known as diabetic encephalopathy, which is well characterized by cellular, molecular and functional changes in the brain of diabetic subjects and rodents. However, little is known about the mechanism of damages and the therapeutic strategies in ameliorating those damages in the diabetic brain. In this study, we utilized a flavonoid, morin which is emerging as a potent drug against a wide range of free radical-mediated as well as neurodegenerative diseases. Morin (15 and 30 mg/kg body weight/day) was orally administered to two different groups of rats after 1 week of diabetes induction, and continued for five consecutive weeks. Two other untreated groups of diabetic and non-diabetic rats were used to compare with drug-treated groups. After drug treatments, cerebral cortex of the brain harvested and analyzed for different factors. Morin supplementation especially at high dose increased the levels of insulin, reduced glutathione, superoxide dismutase and catalase activities, and decreased fasting glucose and thiobarbituric acid reactive substances in the diabetic brain compared to untreated diabetic rats (P < 0.05). Morin also significantly decreased the level of inflammatory markers (TNFα, IL1β, IL-6) in the diabetic brain compared to untreated diabetic rats. Furthermore, the drug influenced an increase in the level of neurotrophic factors (BDNF, NGF and IGF-1) in the diabetic brain compared to untreated diabetic rats (P < 0.05). Thus, our results indicate a beneficial effect of morin by decreasing oxidative stress, inflammation and increasing the neurotrophic support in the diabetic brain, which may ameliorate diabetic encephalopathy.

Rosmarinic acid prevents lipid peroxidation and increase in acetylcholinesterase activity in brain of streptozotocin‐induced diabetic rats

We investigated the efficacy of rosmarinic acid (RA) in preventing lipid peroxidation and increased activity of acetylcholinesterase (AChE) in the brain of streptozotocin-induced diabetic rats. The animals were divided into six groups (n = 8): control, ethanol, RA 10 mg/kg, diabetic, diabetic/ethanol and diabetic/RA 10 mg/kg. After 21 days of treatment with RA, the cerebral structures (striatum, cortex and hippocampus) were removed for experimental assays. The results demonstrated that the treatment with RA (10 mg/kg) significantly reduced the level of lipid peroxidation in hippocampus (28%), cortex (38%) and striatum (47%) of diabetic rats when compared with the control. In addition, it was found that hyperglycaemia caused significant increased in the activity of AChE in hippocampus (58%), cortex (46%) and striatum (30%) in comparison with the control. On the other hand, the treatment with RA reversed this effect to the level of control after 3 weeks. In conclusion, the present findings showed that treatment with RA prevents the lipid peroxidation and consequently the increase in AChE activity in diabetic rats, demonstrating that this compound can modulate cholinergic neurotransmission and prevent damage oxidative in brain in the diabetic state. Thus, we can suggest that RA could be a promising compound in the complementary therapy in diabetes.

Erratum to: Impaired amyloid β-degrading enzymes in brain of streptozotocin-induced diabetic rats

Erratum to: Impaired amyloid β-degrading enzymes in brain of streptozotocin-induced diabetic rats

Impaired amyloid β-degrading enzymes in brain of streptozotocin-induced diabetic rats

Enzymes that degrade the amyloid β-peptide (Aβ) are important regulators of cerebral Aβ levels. High level of Aβ was found in the brain of diabetic patients and diabetic animals. Aim of the study was to investigate whether activities of Aβ-degrading enzymes neprilysin (NEP), endothelin-converting enzyme 1 (ECE-1) and insulin-degrading enzyme (IDE) were impaired in the brain of diabetic rats. Diabetes was induced in rats by ip administration of 65 mg/kg streptozotocin. The temporal cortex and hippocampus were obtained for activity and mRNA level assays of the three enzymes on the 35th day after induction. ECE-1 activity was significantly decreased both in the hippocampus and cortex of diabetic rats, while for IDE significantly lower activity occurred only in the cortex. NEP activity was slightly decreased in both brain regions. The hippocampus of diabetic rats showed significant decrease in mRNA levels of NEP and ECE-1 and moderate increase in IDE mRNA level. The cortex of diabetic rats showed slight decrease in mRNA levels of the three enzymes. The results indicated that the three Aβ-degrading enzymes were damaged to different extents in the brain of diabetic rats, and impairment of ECE-1 and IDE partly contributed to the elevated Aβ(1-40) levels in brain of diabetic rats.

Increased amyloid β-peptide (1–40) level in brain of streptozotocin-induced diabetic rats

The aims of the study were to investigate whether the level of amyloid β-peptide (Aβ) (1–40) was increased in brain of diabetic rats and whether the increase was associated with dysfunction of P-glycoprotein at the blood–brain barrier. A diabetes-like condition was induced by single administration of 65 mg/kg streptozotocin via i.p. injection. Aβ (1–40) levels in brain of the diabetic rats were measured using an enzyme linked immunosorbent assay (ELISA) kit. The in vivo brain-to-blood efflux and blood-to-brain influx transport of [ 125I]-labeled human amyloid-β-peptide (hAβ) (1–40) were measured using the brain efflux index and brain permeability coefficient-surface area product, respectively. [ 14C]inulin served as a reference compound. The results showed that Aβ (1–40) levels significantly increased in temporal cortex and hippocampus of the diabetic rats. The brain remaining percentage of [ 125I]hAβ (1–40) in diabetic rats significantly increased at 30 min after intracerebral microinjection, accompanied by decrease of the brain efflux index. Pretreatment of P-glycoprotein inhibitors verapamil or cyclosporin A significantly increased the brain remaining percentage of [ 125I]hAβ (1–40). The brain permeability coefficient-surface area product of [ 125I]hAβ (1–40) was increased in diabetic rats, accompanied by increased Aβ (1–40) levels in plasma. The present study demonstrated that a diabetic state could increase Aβ (1–40) levels in brain, which might be explained, at least in part, by the decline in brain-to-blood efflux of Aβ (1–40) due to deficient cerebral P-glycoprotein function in diabetic rats.

Functional Induction of P-glycoprotein in the Blood-Brain Barrier of Streptozotocin-Induced Diabetic Rats: Evidence for the Involvement of Nuclear Factor-κB, a Nitrosative Stress-Sensitive Transcription Factor, in the Regulation

The objective of this study was to investigate the transport kinetics of cyclosporin A, a well known substrate for P-glycoprotein (P-gp), across the blood-brain barrier (BBB), and the expression of the transporter in the brain of streptozotocin-induced diabetic rats. The in vivo transport clearance of cyclosporin A was significantly reduced in diabetic rats compared with that in the control. The decreased transport was associated with the increased level of mRNA and the protein for P-glycoprotein in the rat brain. The functional activity of the efflux transporter in mouse brain capillary endothelial (MBEC4) cells, an in vitro model of the BBB, was also stimulated when slow nitric oxide (NO)-releasing donors were present, whereas the stimulation was absent in the case of rapid NO-releasing donors (e.g., S-nitroso-N-acetyl-dl-penicillamine and diethylenetriamine). The stimulatory effect was highest for sodium nitroprusside (SNP) and the functional induction associated with the increased mRNA and protein level of the transporter. The pretreatment of the cell with SNP along with ascorbate, methylene blue, or superoxide dismutase attenuated the induction of function and expression for P-glycoprotein, suggesting that the reaction product between superoxide and NO is involved in the induction of function and expression. The level of nuclear translocation of nuclear factor-kappaB (NF-kappaB) and DNA binding activity of nuclear extracts to the NF-kappaB consensus oligonucleotide was increased in MBEC4 cells pretreated with SNP. Taken together, these observations suggest that nitrosative stress leads to the up-regulation of the message for the efflux transporter and, ultimately, to the enhanced function, probably via a NF-kappaB-dependent mechanism.

Differential effect of insulin treatment on decreased levels of beta-defensins and Toll-like receptors in diabetic rats

Many infections are associated with diabetes, as the ability of the body to fight pathogens is impaired. Recently, altered levels of defensins and polymorphism in Toll-like receptors (TLRs), two crucial components of the innate immune system, have been associated with diabetes. To study the functionality of the innate immune system during diabetes, we measured the expression levels of rat beta-defensin 1 (rBD-1), rBD-2, rTLR2, and rTLR4 in the kidney, lung, liver, and brain of streptozotocin-induced diabetic rats with and without insulin treatment. In the kidneys of diabetic rats, lower levels of rBD-1, rTLR2, and rTLR4 were found. These low levels could be restored with insulin treatment. In contrast, rBD-2 was highly induced in the lungs of diabetic rats, most likely, as a result of a pro-inflammatory response. In the liver and brain of diabetic rats, lower levels of TLRs and/or defensins could not be restored with insulin treatment. Our findings suggest for the first time that high blood glucose and/or insulin deficiency abrogates the expression level of the innate immune system components, defensins and TLRs, explaining the recurrent infections during diabetes. These findings may lay the grounds for future design of novel treatments that will induce defensins and TLRs and, as a result, bolster the immune system in diabetic patients.

Ameliorative effects of dietary caloric restriction on oxidative stress and inflammation in the brain of streptozotocin-induced diabetic rats

Background There is a strong association between oxidative stress and inflammation in the pathologies of diabetes. Recent evidence suggests that these phenomena could impair brain function. We investigated the potential role of dietary caloric restriction in ameliorating the effects of both oxidative stress and inflammation in the brain of streptozotocin-induced type 2 diabetic rats. Methods Forty male Wistar rats were subjected to 30% caloric restriction (20 animals) and ad libitum feeding (20 animals) for 9 weeks before the induction of diabetes in 20 animals (10 from each group) by intraperitoneal injection of 35 mg/kg body weight streptozotocin. Results Caloric restriction was able to significantly ( p > 0.05) reduce triglyceride, ROS, IL6, TNF-α and body weights in diabetic rats. However, no significant differences were obtained in the antioxidant enzyme (SOD, CAT and GPx) activities except in GPx where caloric restriction increased the levels in both non-diabetic and diabetic rats. Conclusion Caloric restriction was found to ameliorate the oxidative and inflammatory effects of diabetes in the brain. Non-diabetic rats feeding ad libitum were found to have increased levels of oxidative stress and inflammatory biomarkers and these could, in part, be due to their increased body weights.

Effects of vitamin E on oxidative stress and membrane fluidity in brain of streptozotocin-induced diabetic rats

Background: Diabetics and experimental animal models exhibit high oxidative stress due to persistent and chronic hyperglycemia, thereby deplete the activity of the antioxidative defense system and thereby promote the generation of free radicals. The current study examined the effects of vitamin E on oxidative stress and membrane fluidity in the brain of diabetes-induced rats. Methods: Sprague–Dawley male rats were randomly assigned to normal and streptozotocin (STZ)-induced diabetic groups. The diabetic groups were fed a vitamin E-free diet, 40 mg vitamin E/kg diet, or 400 mg vitamin E/kg diet. Diabetes was induced with STZ after 3 weeks of the experimental diet, then the rats were sacrificed 9 days later to determine the oxidative stress and cell membrane fluidity in the brain. Results: Dietary vitamin E strengthened the antioxidative defense system with an increased activity of the antioxidant enzymes, such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) and increased vitamin E content, in the brain of the diabetes-induced exeperimental rats. Accordingly, vitamin E was found to reduce the accumulation of reactive oxygen species (ROS), such as superoxide radical decrease the generation of oxidative damage substances, such as the carbonyl value, increase the membrane fluidity lowered by oxidative damage, and significantly improve the lipid composition. Conclusions: Vitamin E was found to be excellent for strengthening the antioxidative defense system, reducing the generation of ROS and damaging oxidative substances, and maintaining membrane fluidity in the brain of diabetes-induced rats.

Effect of rosiglitazone on the differential expression of diabetes-associated proteins in pancreatic islets of C57Bl/6 lep/lep mice.

The insulin sensitizer drug, rosiglitazone, has been shown to have a protective effect on pancreatic islet cell structure and function in animal models of type 2 diabetes. The identification of new molecular targets associated both with islet cell dysfunction and protection is a crucial research goal. In the present study, a proteomics approach has been used to identify such targets. Obese C57Bl/6J lep/lep mice and lean littermates were given the insulin sensitizer drug BRL49653, rosiglitazone. It normalized the impaired glucose tolerance in lep/lep mice but had no significant effect on glucose tolerance in the lean mice. Pancreatic islet polypeptides were arrayed by a two-dimensional gel electrophoresis system that separated more than 2500 individual spots. Three overexpressed and six underexpressed proteins were significant (p < 0.05) between lep/lep and lean mice, and four were modulated significantly (p < 0.05) by the rosiglitazone treatment of the obese mice. The identity of these differentially expressed proteins was made using mass spectrometric analysis and provided evidence that differential expression of actin-binding proteins may be an important aspect of defective islet function. Rosiglitazone increased carboxypeptidase B expression in both lep/lep and normal mice suggesting that this might be an independent effect of rosiglitazone that contributes to improved insulin processing.

Changes of superoxide dismutase gene expression and activity in the brain of streptozotocin-induced diabetic rats

In an attempt to discover the changes of superoxide dismutase (SOD) in the brain of diabetic state, we investigated the level of SOD in streptozotocin-induced diabetic rats (STZ-diabetic rats) using enzyme activity assay, Northern blotting analysis of mRNA levels and Western blotting of enzyme amount. Five discrete brain regions, cerebrocortex, hypothalamus, hippocampus, the remaining non-cortex cerebrum (NCC area) and cerebellum, were examined in STZ-diabetic rats to compare with age-matched normal rats. Higher levels of Mn-SOD including the activity, mRNA expression and immunoblot of enzyme were found in all areas of the brain from STZ-diabetic rats as compared with that in Wistar rats. Except in the cerebellum, similar changes of Cu,Zn-SOD were found in the brain of STZ-diabetic rats. These results indicate an increase of SOD both gene expression and activity in the brain of STZ-diabetic rats. This alteration of SOD may be one of the important factors for the vulnerability of the brain to oxygen free radicals or may be related to the pathophysiology of diabetes.

Structural and functional consequences of increased tubulin glycosylation in diabetes mellitus.

The extent of in vitro nonenzymatic glycosylation of purified rat brain tubulin was dependent on time and glucose concentration. Tubulin glycosylation profoundly inhibited GTP-dependent tubulin polymerization. Electron microscopy and NaDodSO4/polyacrylamide gel electrophoresis showed that glycosylated tubulin forms high molecular weight amorphous aggregates that are not disrupted by detergents or reducing agents. The amount of covalently bound NaB3H4-reducible sugars in tubulin recovered from brain of streptozotocin-induced diabetic rats was dramatically increased as compared with tubulin recovered from normal rat brain. Moreover, tubulin recovered from diabetic rat brain exhibited less GTP-induced polymerization than tubulin from nondiabetic controls. The possible implications of these data for diabetic neuropathy are discussed.