Alloxan-induced Mice Research Articles

Protective effect of vitamin E against alloxan-induced mouse hyperglycemia

BackgroundAlloxan induces oxidative stress and hyperglycemia in animal models. Acatalasemic (catalase deficiency) mice are susceptible to alloxan-induced hyperglycemia. As the incidence of hyperglycemia induced by alloxan was reportedly improved when mice were fed a vitamin E supplemented diet, this protective effect was examined. MethodsAcatalasemic and normal mice fed a vitamin E supplemented diet were treated with alloxan. The pancreas were examined with microscopy. We also isolated pancreatic islets of normal mice treated with alloxan. The glucose stimulated insulin secretion was examined. ResultsVitamin E powerfully ameliorated the increase in apoptosis. Vitamin E increases insulin amounts secreted from pancreatic cells, but does not ameliorate the regulation of the glucose stimulated insulin secretion. ConclusionsIt is suggested that the difference in the mice fed vitamin E supplemented diet is due to an increase of insulin secretion and that vitamin E supplementation may have a role in helping to slow the stages of diabetes mellitus.

Timosaponin B-II ameliorates diabetic nephropathy via TXNI P, mTOR, and NF-κB signaling pathways in alloxan-induced mice

BackgroundMany synthesized drugs with clinical severe side effects have been used for diabetic nephropathy (DN) treatment. Therefore, it is urgent and necessary to identify natural and safe agents to remedy DN. Timosaponin B-II (TB-II), a major steroidal saponin constituent in Anemarrhena asphodeloides Bunge, exhibits various activities, including anti-inflammatory and hypoglycemic functions. However, the anti-DN effects and potential mechanism(s) of TB-II have not been previously reported.PurposeTo investigate the effect of TB-II on DN in alloxan-induced diabetic mice.MethodsTB-II was isolated and purified from A. asphodeloides Bunge using macroporous adsorption resin and preparative high-performance liquid chromatography. The effect of TB-II on DN was evaluated in alloxan-induced diabetic mice using an assay kit and immunohistochemical determination in vivo. The expression of mammalian target of rapamycin (mTOR), thioredoxin-interacting protein (TXNIP), and nuclear transcription factor-κB (NF-κB) signaling pathways was also measured using Western blot analysis.ResultsTB-II significantly decreased the blood glucose levels and ameliorated renal histopathological injury in alloxan-induced diabetic mice. In addition, TB-II remarkably decreased the levels of renal function biochemical factors, such as kidney index, blood urea nitrogen, serum creatinine, urinary uric acid, urine creatinine, and urine protein, and it reduced lipid metabolism levels of total cholesterol and triglycerides and the levels of inflammatory cytokines interleukin-6 and tumor necrosis factor-α in alloxan-induced mice. Furthermore, TB-II inhibited the expression of mTOR, TXNIP, and NF-κB.ConclusionThe results revealed that TB-II plays an important role in DN via TXNIP, mTOR, and NF-κB signaling pathways. Overall, TB-II exhibited a prominently ameliorative effect on alloxan-induced DN.

Effect of Burdock Root and the Fermented Product on Alloxan-Induced Mouse Hyperglycemia

<p><strong>Backgrounds:</strong> We reported that feeding 5% <em>Asperagillus awamori-</em>fermented burdock root diet was effective in preventing mouse hyperglycemia caused by alloxan.</p> <p><strong>Methods:</strong> Diets containing 5% burdock roots were prepared from raw and <em>Asperagillus awamori-</em>fermented burdock root powders. Acatalasemic mice<strong>,</strong> having a quite low catalase activity in blood, and normal mice were fed these diets for 14 weeks, separately. Then, alloxan (200 mg/ kg of body weight) or PBS was intraperitoneally administrated to each mouse. After 5 day from the administration, blood glucose assay and glucose tolerance test were carried out, and then insulin, C-peptide and lipid peroxide in plasma were examined.</p> <p><strong>Results:</strong> Incidences of hyperglycemia in normal mice fed control, raw and fermented burdock root diets were 25, 20 and 11 %, respectively, and these in acatalasemic mice<strong> </strong>were 73, 80 and 27%. Insulin and C-peptide in plasma of mice fed raw burdock root diet or control diet were low compared to mice fed the fermented diet.</p> <p><strong>Conclusions:</strong> Intake of raw burdock root does not suppress the alloxan-induced hyperglycemia but the fermented burdock root does. It is suggested that <em>Asperagillus awamori</em> plays an important role for the prevention.</p>

Hypoglyceamic Effect of Methanolic Extract of Pergularia daemia in Alloxan-Induced Diabetic Mice

Objective: The aim of the present study was to investigate the antidiabetic potential of Pergularia daemiamethanolic extract (PD) on alloxan-induced diabetic mice.Methods: Diabetes was induced using alloxan (150mg/kg). Different doses of the methanolic extract of P. daemia were prepared and administered orally. The alloxan-induced diabetic mice were treated with 200, 400 and 1000 mg/kg body weight (b.w.) methanolic extract of Pergularia daemia and the blood glucose levels were measured over 4 hrs.Results: All doses of the extract (200, 400 and 1000 mg/kg b.w) significantly (p<0.0001) lowered fasting blood glucose level after treatment. The hypoglycaemic effect of the methanolic extract was also compared with that of an oral dose of glibenclamide under the same conditions. The extract showed significant (p<0.05) hypoglycemic effect in alloxan-induced diabetic mice. The glucose half-life (t1/2G) (p<0.0001), Mean Glucose Residence Time (MRTG) (p<0.0001), and the area under glucose concentration time curve (AUC0-4hG) (p<0.05) were high in control group compared to the alloxan-induced mice treated with P. daemia extract.Conclusion: The results of this study indicate that methanolic extract of the plant of Pegularia daemiapossess hypoglycaemic activity.

Effect of vitamin E on alloxan-induced mouse diabetes

ObjectivesAlloxan generates hydrogen peroxide in the body, and a small amount of alloxan administered to acatalasemic mice results in diabetes. D-α-Tocopherol (vitamin E) is an antioxidant which helps prevent excess oxidation in the body. In this study, we examined the effect of vitamin E on diabetes caused by alloxan administration in mice. MethodsMice were maintained on a vitamin E-deprived diet and supplemented diet, respectively, for 14weeks. Alloxan was then intraperitoneally administered, and blood glucose, glucose tolerance and the insulin level in mouse blood were examined. ResultsHyperglycemia was observed in the mice maintained on the vitamin E-deprived diet. The incidence of hyperglycemia in the mice maintained on the vitamin E-deprived diet was significantly higher than that in the mice maintained on the supplemented diet. The abnormal glucose metabolism caused by alloxan administration was ameliorated by the vitamin E-supplemented diet. ConclusionsIt is deduced that vitamin E can prevent a decrease of insulin concentration in the blood in this mouse model.

Methanolic Root Extract of Rauwolfia serpentina Benth Improves the Glycemic, Antiatherogenic, and Cardioprotective Indices in Alloxan-Induced Diabetic Mice.

The aim of the study was to evaluate the phytochemistry and the effect of methanolic root extract (MREt) of Rauwolfia serpentina on alloxan-induced diabetic Wister male mice. Mice were divided in control (distilled water at 1 mL/kg) and alloxan-induced diabetic mice which subdivided into diabetic (distilled water at 1 mL/kg), negative (0.05% dimethyl sulfoxide at 1 mL/kg), positive (glibenclamide at 5 mg/kg) controls, and three test groups (MREt at 10, 30, and 60 mg/kg). All treatments were given orally for 14 days. Qualitatively MREt showed the presence of alkaloids, carbohydrates, flavonoids, glycosides, cardiac glycosides, phlobatannins, resins, saponins, steroids, tannins, and triterpenoids, while quantitatively extract was rich in total phenols. The flavonoids, saponins and alkaloids were also determined in root powder. MREt found effective in improving the body weights, glucose and insulin levels, insulin/glucose ratio, glycosylated and total hemoglobin in test groups as compared to diabetic control. Similarly, significantly decreased levels of total cholesterol, triglycerides, low-density lipoprotein (LDL-c), and very low-density lipoprotein (VLDL-c) cholesterols were found in test groups. Significant lipolysis with improved glycogenesis was also found in liver tissues of all test groups. ALT levels were found normal in all groups. Thus, MREt improves the glycemic, antiatherogenic, coronary risk, and cardioprotective indices in alloxan-induced diabetic mice.

Protective effect of Pedicularis decora Franch root extracts on oxidative stress and hepatic injury in alloxan-induced diabetic mice

Pedicularis decora (P. decora) has been known to have antioxidative effect, but the effect of its successive extracts has not been well studied. Our objective was to investigate the effect of n-butanol and water extracts of P. decora Franch root on the oxidative stress and hepatic injury in alloxan-induced diabetic mice. Diabetes was induced in ICR mice by a single alloxan injection (150 mg kg−1 b. w., i.p.). After diabetes was induced, the mice were treated with n-butanol and water extracts of P. decora Franch root, α-tocopherol and saline for three weeks. Treatment with α-tocopherol, n-butanol and water extract of P. decora Franch root caused considerable gain in body and liver weight, glucose, liver glycogen and serum insulin concentration. The two extracts of P. decora Franch and α-tocopherol were found to show hepatoprotective effect by lowering down the contents of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), cholesterol and triglyceride and elevating HDL cholesterol in serum in alloxan induced diabetic mice. Treatment with the two extracts of P. decora Franch and α-tocopherol significantly lowered oxidative stress biomarkers such as lipid peroxidation (LPO) levels in serum and in liver tissue, nitric oxide (NO) levels in serum and catalase (CAT) activities in liver tissue against alloxan treated mice. Reduced glutathione (GSH) and hepatic enzymes like superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-s-transferase (GST) as well as serum total antioxidant activity (TAA) were significantly increased by treatment with the two extracts and α-tocopherol, against alloxan treated mice. Based on these results, it was observed that α-tocopherol, n-butanol and water extract of P. decora Franch root administration could counteract alloxan induced the diabetic symbols and inhibit oxidative stress and hepatic injury. Additionally, n-butanol extract was more pronounced on the parameters studied of the oxidative stress and hepatic injury. The n-butanol extract is more protective than water extract suggests that n-butanol extract may better ameliorate the harmful effects in alloxan-induced mice. Key words: Pedicularis decora Franch, n-butanol extract, water extract, hepatic injury, oxidative stress.

HYPOGLYCEMIC EFFECT OF SEMIPURIFIED PEPTIDES FROM MOMORDICA CHARANTIA L. VAR. ABBREVIATA SER. IN ALLOXAN-INDUCED DIABETIC MICE

ABSTRACT Hypoglycemic effect of semipurified peptides from an aqueous extract of Momordica charantia L. var. abbreviata Ser. (MCV) in alloxan-induced diabetic mice was investigated. MC2-1, a semipurified peptide fraction from MCV produced a significant hypoglycemic effect in diabetic mice when compared to the diabetic control group. In a long-term study, mice were treated with MC2-1 and glibenclamide, respectively, once a day, for 28 days. Repeated oral administration of MC2-1 reduced significantly the fasting blood glucose level during the 28 days of treatment. Furthermore, MC2-1 significantly reduced hepatic malondialdehyde level, glycogen content, serum cholesterol, triglyceride and low-density lipoprotein cholesterol levels. At the same time, MC2-1 markedly increased high-density lipoprotein cholesterol and serum insulin levels after 28 days of treatment in diabetic mice. Loss in body weight was also prevented in diabetic mice. Hence, MC2-1 can be incorporated as a supplement in health-care food, drugs and/or combined with other hypoglycemic drugs. PRACTICAL APPLICATIONS The present study showed that peptide fraction MC2-1 obtained from an aqueous Momordica charantia L. var. abbreviata Ser. (MCV) extract has a strong hypoglycemic effect in alloxan-induced mice. MC2-1 administered at a dose of 20 mg/kg has a similar or even stronger hypoglycemic effect as that produced by the MCV aqueous extract administered at a dose of 500 mg/kg. The study revealed that MC2-1 is the main fraction with hypoglycemic effect in MCV aqueous extract. Through the removal of compounds with little or no hypoglycemic effect, the dosage needed for diabetes prevention and control is significantly reduced. At the same time, the compounds present in the aqueous extract, which could affect blood glucose level lowering effect, are minimized. MC2-1 can be added as supplement in health-care food, drugs or combined with other hypoglycemic drugs. In addition, the present study revealed the mechanism of action of the hypoglycemic fraction.

Isolation and intracellular localization of insulin-like proteins from leaves of Bauhinia variegata

Evidence based on immunological cross-reactivity and anti-diabetic properties has suggested the presence of insulin-like peptides in plants. The objective of the present study was to investigate the presence of insulin-like proteins in the leaves of Bauhinia variegata ("pata-de-vaca", "mororó"), a plant widely utilized in popular medicine as an anti-diabetic agent. We show that an insulin-like protein was present in the leaves of this plant. A chloroplast protein with a molecular mass similar to that of bovine insulin was extracted from 2-mm thick 15% SDS-PAGE gels and fractionated with a 2 x 24 cm Sephadex G-50 column. The activity of this insulin-like protein (0.48 mg/mL) on serum glucose levels of four-week-old Swiss albino (CF1) diabetic mice was similar to that of commercial swine insulin used as control. Further characterization of this molecule by reverse-phase hydrophobic HPLC chromatographic analysis as well as its antidiabetic activity on alloxan-induced mice showed that it has insulin-like properties. Immunolocalization of the insulin-like protein in the leaves of B. variegata was performed by transmission electron microscopy using a polyclonal anti-insulin human antibody. Localization in the leaf blades revealed that the insulin-like protein is present mainly in chloroplasts where it is also found associated with crystals which may be calcium oxalate. The presence of an insulin-like protein in chloroplasts may indicate its involvement in carbohydrate metabolism. This finding has strengthened our previous results and suggests that insulin-signaling pathways have been conserved through evolution.

Neopterin as an endogenous antioxidant

The in vitro potency of neopterin (NP) as an antioxidant and its in vivo activity to suppress alloxan-induced diabetes were investigated. The reduced form of neopterin, 5,6,7,8-tetrahydroneopterin (NPH-4), showed an extremely high superoxide anion radical scavenging activity in two assay systems, i.e. xanthine/xanthine oxidase- and macrophage/phorbol myrislate acetate (PMA)-reaction systems. NPH-4 also inhibited the oxidation of linoleic acid about as effectively as uric acid. Furthermore, NPH-4 and NP effectively suppressed alloxan-induced mouse diabetes. These results suggest that pteridines play an important role as endogenous antioxidant.

血清胸腺因子（ＦＴＳ）によるマウス・アロキサン糖尿病に対する抑制効果とリポソームによる薬効制御

The suppressive effects of facteur thymique serique (FTS) on alloxan-induced mouse diabetes were investigated in a detailed manner. The elevation of blood glucose was suppressed at a dose of 1 μg to 100 μg per animal in a biophasic manner. The timing of FTS injection was effective only within 5 min before alloxan treatment. The administration of and-FTS antibody prevented the suppressive effect of FTS on alloxan-induced diabetes. The clearance of FTS from blood circulation was significantly fast with a calculated initial half life of 2 to 3 min following i.v injection. The frequent administration of FTS would be necessary due to the rapid clearance of it from circulation. So, a potential sustained-release drug delivery system was developed by using liposomes. Liposomes containing gangliosides GM1 (GM1-LP) were effective in extending the timing of FTS pretreatment. These results suggest that liposomal FTS might be applicable as a therapeutic agent for diabetes.

Isolation and hypoglycaemic activity of panaxans A, B, C, D and E, glycans of Panax ginseng roots.

An aqueous methanol/water extract of the Oriental crude drug “ninjin” (ginseng), PANAX GINSENG roots from Korea, showed a remarkable hypoglycaemic activity on administration to mice. Fractionation of the extract, along with the pharmacological assays, led to isolation of five glycans, panaxans A, B, C, D and E. These constituents exhibited significant hypoglycaemic actions in normal and alloxan-induced hyperglycaemic mice.