Methods to Reduce the Hypoglycemic Mortality of Alloxan in Diabetic Rabbit Model

Objective: To explore an intervention method to reduce the mortality of alloxan diabetes model, and to preliminarily analyze the mechanism of alloxan induced animal death. Methods: Healthy New Zealand rabbits were randomly divided into injection group, control group, experimental group and blank group. The single injection group was injected with 100 mg/kg alloxan once. The control group was given 5% glucose solution and 100 mg/kg alloxan was injected in two times. The experimental group was given 5% glucose solution orally, 100 mg/kg alloxan, 7 mL 0.9% NaCl intravenously and 5 mL 5% glucose intraperitoneally immediately, and blood glucose was continuously monitored, 10 mL 5% glucose intravenously and 10 mL 5% glucose intraperitoneally every 4 h in the hypoglycemic stage. The blank group does nothing. Liver and kidney tissues at different time periods were stained with HE and organ index was evaluated. Results: 1) A single injection of 100 mg/kg alloxan without any intervention resulted in 100% mortality. Before modeling, oral administration of 5% glucose solution, divided into two injections of 100 mg/kg alloxan, mortality reached 100%;A single injection of 100 mg/kg alloxan and continuous intervention of normal saline and glucose for 20 h can significantly reduce the mortality of alloxan induced diabetic rabbit model. 2) Liver and kidney tissues were damaged in different degrees at different time periods, and liver and kidney indexes were significantly increased after alloxan injection compared with the normal group, with statistical significance (P > 0.05). Conclusion: 1) Every 4 hours of hypoglycemia, 10 ml 5% glucose was injected intravenously 10 ml 5% glucose intraperitoneally. It can reduce the death rate of alloxan diabetic rabbit model and shorten the time of blood glucose measurement. 2) After the injection of alloxan, acute lesions of liver and kidney may occur in different degrees, or one of the causes of acute death of experimental animals.

STZ与ALLOXAN诱导大鼠糖尿病模型胰腺的组织学观察

目的 比较STZ和ALLOXAN诱导糖尿病的效果 ,为实验性糖尿病模型的建立提供实验依据。方法 对 6 0只SD大鼠分别采用一次性腹腔注射STZ和ALLOXAN建立大鼠糖尿病模型 ,并监测血糖、尿糖的变化以及对胰腺组织进行免疫学染色观察。结果 注射化学药物 4 8小时后血糖值达到成模标准者 ,STZ为 96 6 % ,ALLOXAN为 93 3% ;对两种模型大鼠的胰岛进行组织学及免疫染色观察 ,二者无明显区别。结论 ALLOXAN在腹腔注射诱导大鼠糖尿病模型上可以替代STZ ,尤其适用于大批模型的建立及胰岛再生功能的研究。

Alloxan糖尿病模型复制中的多因素作用

用Ａｌｌｏｘａｎ复制大鼠糖尿病模型受诸多因素的影响。本实验结果提示：动物饲料成分；腹腔注射Ａｌｌｏｘａｎ剂量、次数；动物体重；个体差异均可对模型复制的稳定性产生影响，并与血糖变化及动物死亡数有密切关系。

Adrenocortical system activity in alloxan-resistant and alloxan-susceptible Wistar rats

In the dynamics of the disease development, diuresis and glycosuria increase in alloxan-susceptible rats, while in alloxan-resistant rats the increase in the values of these indices is expressed to a lesser extent, and they begin to decrease by day 8 of the disease. In alloxan-susceptible rats, the mass index of adrenal gland is increased, and that of thymus is decreased and corticosterone concentration in blood, adrenal gland and urine as well as alanine and aspartate aminotransferase activities in liver are increased;in alloxan-resistant rats the values of these indices do not differ from those of rats of the control group.

Alloxan诱发糖尿病实验大鼠微量元素谱的多元分析

为确认Ａｌｌｏｘａｎ诱发糖尿病鼠与微量元素的关连，用多元分析研究了糖尿病鼠样品中微量元素，并引出对糖尿病的新见解。用ＩＣＰ－ＡＥＳ测定了Ａｌｌｏｘａｎ诱发糖尿病鼠和对照鼠的骨、心、肾、脾、肺、肝、血和毛中１９种微量和宏量元素（Ａｌ、Ａｓ、Ｂ、Ｂａ、Ｃｄ、Ｃｏ、Ｃｒ、Ｍｇ、Ｍｎ、Ｎｉ、Ｐｂ、Ｓｅ、Ｚｎ、Ｆｅ、Ｃｕ、Ｃａ、Ｓｒ、Ｔｉ、Ｖ），结果在糖尿病鼠与对照组的微量元素间呈现一些显著差异。多元分析找到一条共同相关链（Ｍｎ－Ｎｉ－Ｃｕ－Ｓｒ－Ｔｉ），血、发样相同，与样品种类无关，此链提供了关于机体的某些特殊信息。经非线性映射，各组映射象图显示出糖尿病和对照组的样品点分布在图上不同区域。

Antihyperglycemic and antioxidant effects of Solanum xanthocarpum leaves(field grown & in vitro raised) extracts on alloxan induced diabetic rats

Objective:To investigate antidiabetic efficacy of the extract of field grown and in vitro raised leaves of Solanum xanthocarpum(S.xanthocarpum) against alloxan induced diabetic rats. Methods:The antidiabetic activity of the crude methanol extracts of the field grown and in vitro raised leaves of S.xanthocarpum at different concentrations(100-200 mg/kg bw) was tested against alloxan induced diabetic rats.The antidiabetic efficacy was validated through various biochemical parameters and the antioxidant effect was also determined.The phytochemical analyses of field grown S.xanthocarpum and in vitro rasied 5.xanthocarpum leaves were done by estimating their chlorophyll,carotenoids,total sugar,protein,amino acid and minerals contents. Results:The results revealed that the methanol extracts of both the leaves(field grown and in vitro raised) of S.xanthocarpum was efficient anti hyperglycemic agents at a concentration of 200 mg/kg bw and posses potent antioxidant activity.However,the extracts of in vitro rasied S.xanthocarpum raised leaves exhibit higher efficacy than the field grown leaves in all tested concentrations.Proximal composition and mineral analysis of S.xanthocarpum revealed higher concentration of contents in in vitro rasied S.xanthocarpum than field grown S.xanthocarpum. Conclusions:From the results it can be concluded that the leaves extracts of S.xanthocarpum can be a potential candidate in treating the hyperglycemic conditions and suits to be an agent to reduce oxidative stress.

Antidiabetic activity of methanolic bark extract of Alhizia odoratissima Benth.in alloxan induced diabetic albino mice

Objective:To evaluate the antidiabetic potential of methanolic extract of Albizia odoratissima Benth.bark in alloxan induced diabetic mice.Methods:Group-Ⅰ(normal control) mice received only basal diet without any treatment.In Group-Ⅱ(Diabetic control) mice,diabetes was induced by alloxan(150 mg/kg i.p.) and received only Tween 80.5%v/v in normal saline. Group-Ⅲand Group-Ⅳmice received metformin(10 mg/kg) and gliclazide(10 mg/kg) as standard drugs.Group-ⅤandⅥmice received methanolic bark extract of Albizia odoratissima at doses of 250 and 500 mg/kg body weight p.o.,respectively.Results:The results of the study indicates that Albizia odoratissima bark extract significantly(P【0.01) reduced the blood sugar level.The bark extract also significantly reduced the levels of serum cholesterol,triglycerides, serum glutamic-oxaloacetic transaminase,serum glutamic-pyruvic transaminase,alkaline phosphatase and decreases level of total proteins in alloxan induced diabetic mice.Conclusions: Methanolic extract of Albizia odoratissima has protective effects on the protection of vital tissues(pancreas,kidney,liver,heart and spleen),thereby reducing the causation of diabetes in experimental animals.

Evaluation of protective effect of cactus pear seed oil(Opuntia ficusincida L. MILL.) against alloxan-induced diabetes in mice

Objective: To evaluate the in vitro antioxidant power of cactus pear seed oil [Opuntia ficusincida L. MILL.(CPSO)] and its protective effect against chemically induced diabetes mellitus in mice. Methods: The in vitro antioxidant effect of CPSO was evaluated using 2,2-diphenyl-1-picrylhydrazyl(DPPH) scavenging assay. The preventive effect was conducted on Swiss albino mice treated with CPSO(2 m L/kg, per os), before and after a single intraperitoneal alloxan administration(100 mg/kg). Survival rate, body weight and fasting blood glucose were measured and histopathological analysis of pancreas was performed to evaluate alloxaninduced tissue injuries. Results: CPSO exhibited an antioxidant effect in DPPH scavenging assay. Moreover, the administration of CPSO(2 m L/kg) significantly attenuated alloxaninduced death and hyperglycemia(P<0.001) in treated mice. Morphometric study of pancreas revealed that CPSO significantly protected islets of langerhans against alloxan induced-tissue alterations. Conclusions: Based on theses results, CPSO can prevent alloxan-induced-diabetes by quenching free radicals produced by alloxan and inhibiting tissue injuries in pancreatic β-cells.

Pathological changes in the cellular structures of retina and choroidea in the early stages of alloxan-induced diabetes

AIM To investigate the temporal sequence of pathological changes in the cellular structures of retina and choroidea in the early stages of diabetes in laboratory animals.METHODS Experimental type 1 diabetes was modeled by three intraperitoneal injections of an alloxan solution into 30 male nonlinear rats at 16 wk of age. The 30 th and 60 th days from the final alloxan injection were chosen as the endpoints. Light and electron microscopy and morphometric and immunohistochemical studies were performed on histological slices of eyeballs from experimental animals.RESULTS Diabetic disturbances progressed to 60 d of the experiment. Thus, in the retina, a partial destruction of photoreceptors accompanied by interstitial edema was observed. The morphometric analysis revealed a reduction in the thickness of the retina. A reduction in the number of blood vessels of the choroid with disturbances of the endothelial cells and the vascular walls and a persistent reduction in the number of melanocytes were observed. The number of proliferating Ki-67 positive cells decreased, and the number of macrophages increased with diabetes development.CONCLUSION The starting point in the development of destructive changes involves early reduction in the number of melanocytes of the choroidea and alterations in the retinal pigment epithelium.

Histological changes and antidiabetic activities of Icacina trichantha tuber extract in beta-cells of alloxan induced diabetic rats

Objective:To investigate the antidiabetic,hypolipidaemic activities and histopathological changes ofIcacina trichantha(I.trichantha)tuber extract in alloxan induced diabetic rats.Methods:In the present study,80%methanol extract of I.trichanthatuber was tested on alloxan induced diabetic rats.They were randomly grouped into control(distilled water and glibenclamide)and experimental(200,400 and 600 mg/kg body weight).Diabetes was induced by a single intraperitoneal injection of 160 mg/kg body weight of alloxan.Blood glucose levels weremeasured using blood glucose test strips with AccuCheck Advantage II glucometer at 1,3,6,and 24 h on the first day and 1 h after treatment on Day 7,14 and 21.Blood samples were collected and centrifuged to separate serum for estimation of lipid profile and other biochemical parameters.Histopathological changes in diabetic rats pancreas were also studied after extract treatment.Results:Daily oral administration ofI.trichanthatuber extract(200,400,and 600 mg/kg bodyweight)and glibenclamide(2 mg/kg)showed beneficial effects on blood glucose level(P<0.01)as well as improving liver,kidney functions and hyperlipidaemia due to diabetes.The extract had a favourable effect on the histopathological changes of the pancreas in alloxan induced diabetes.Conclusions:I.trichanthatuber extracts posses antidiabetic activities as well as improve liver and renal profile and total lipids levels.I.trichanthatuber extracts also have favourable effects to inhibit the histopathological changes of the pancreas in alloxan induced diabetes.

Evaluation of antihyperglycemic activity of methanolic Tecomaria capensis Thunb.(Bignoniaceae) leaves extract in alloxan induced hyperglycemic rats

Objective:To evaluate the antihyperglycemic activity of Tecomaria aipensis(T.capensis) Thunb. (Bignoniaceae) methanolic leaves extract(TCLE) using blood glucose level in normal fasted rats, glucose tolerance test and alloxan induced hyperglycemia models.Methods:TCLE(100,300, 1 000 and 2 000 mg/kg body wt.) was given to rats orally to observe acute toxicity,and observed for 14 d.TCLE 200 and 400 mg/kg,and glibenclamide 0.6 mg/kg were given orally in all models. Results:Results demonstrated that the no mortality was reported even after 14 d.This indicates that the methanol extract is safe up to a single dose of 2 000 mg/kg body weight.TCLE(200 and 400 mg/kg p.o.) exhibited remarkable blood glucose lowering effect in blood glucose level in normal fasted rats,glucose tolerance and alloxan induced hyperglycemia model.Cholesterol and triglyceride also decreased in alloxan induced hyperglycemia model.Condosions:The results of this study exhibites that methanol extract of T.capensis possesses antihypergycemic activity and it may prove to be effective for the treatment of hyperglycemia.

Improvement of Dissolution Rate of Gliclazide Using Solid Dispersions with Aerosil 380 and Its Effect on Alloxan Induced Diabetic Rats

The main objective of this research is to conduct a comprehensive study for enhancing the aqueous solubility of poorly water soluble gliclazide using hydrophilic fumed silica particles (Aerosil&#174;380) and evaluating the influence of silica on drug release profile and pharmacological activity on alloxan induced diabetic rats. Solid dispersions (SD’s) of gliclazide were prepared using solvent evaporation method. The dissolution profiles and solid state characterization of the SD’s prepared were all evaluated. The dissolution rate of gliclazide in the SD’s with fumed silica (weight ratio, 1:1) was approximately 38%, which is about 10 fold higher than that of the pure drug after 30 min. After forming the SD’s, gliclazide changed into an amorphous state, which can infer from differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD). Fourier transform infrared spectroscopy (FTIR) also revealed the formation of weak hydrogen bonding through the interactions between the secondary amine groups of gliclazide and silanol groups of silica particles in the SD’s. The rapid dissolution rate from the SD’s might be attributed to the amorphization of drug, improved specific surface area and wettability than the original drug crystals. Further, we investigated the antidiabetic effects of SD’s of gliclazide in alloxan induced diabetic rats. The SD’s of gliclazide decrease the blood glucose level 64% whereas the conventional gliclazide decreases only 37% in diabetic rats. Lipid profiles, kidney and liver functions are remarkably improved in diabetic rat treated with SD’s of gliclazide than that of conventional gliclazide. These results suggest that SD’s of gliclazide have much more bioavailability and hence are more pharmacologically active than that of conventional gliclazide form.

Comparative study between the effect of <i>Momordica charantia</i>(wild and hybrid variety) on hypoglycemic and hypolipidemic activity of alloxan induced type 2 diabetic long-evans rats

This study was undertaken to evaluate the hypoglycemic and hypolipidemic effect of Momordica charantia (wild and hybrid variety) powder on alloxan induced type 2 diabetic male Long-Evans rats. Oral feeding of the M. charantia powder slightly decreased serum total cholesterol, triglyceride levels and LDL-cholesterol compared with wild, hybrid and standard drug. M. charantia wild variety showed more significant (p M. charantia did not show any significant effect on HDL-cholesterol and liver glycogen. Thus, results of the study prove that the wild variety of M. charantia fruit have potent antidiabetic and antilipidemic property.

Effect of <i>Aspergillus awamori</i>-Fermented Burdock Root on Mouse Diabetes Induced by Alloxan—Prevention of Cell Apoptosis

Root of burdock contains high amounts of dietary fibers and polyphenols. To improve the functional properties, the root was fermented with Aspergillus awamori. Effect of the fermented burdock on alloxan-induced mouse diabetes was examined. A diet containing the 5% fermented burdock powers was prepared to examine effect of the burdock diet on alloxan-induced mouse diabetes. Mice fed the burdock diet and the control diet for 14 weeks. Then, alloxan (200 mg/kg of body weight) was administrated to each mouse. After 5 days from the administration, blood glucose assay and glucose tolerance test were carried out. Incidence of hyperglycemia decreased and the glucose metabolism was improved when mice fed the burdock diet. Insulin, C-peptide, biomarkers of oxidative stress in plasma and apoptosis in pancreas were examined and compared to those obtained from mice fed the control diet. It is deduced that alloxan-induced diabetes is caused to lower insulin concentration. The fermented-burdock diet improves the diabetes and prevents apoptosis in the pancreas.

Effect of methanolic extract of Tetrapleura tetraptera(Schum and Thonn)Taub leaves on hyperglycemia and indices of diabetic complications in alloxan-induced diabetic rats

Objective:To investigate the ameliorative role of Tetrapleura tetraplera(Schum and Thonn)Taub(T.tetraplera)leaf in hyperglycemia with associated conditions like oxidative stress,kidney damage and disorders in lipid metabolism.Methods:Five groups of five rats each intraperitoneally received the following treatment schedules for 7 d:untreated normal control,untreated alloxan-diabetic control,diabetic treated with glibenclamide,normal rats treated with extract(50 mg/kg)and diabetic rats treated with the extract.Evaluations were made for fasting blood sugar,body weight changes,malondialdehyde,aspartate aminotransferase,alanine aminotransferase,bilirubin,superoxide dismutase,catalase,lipid profile,packed cell volume,hemoglobin,urea and creatinine in all the rats.Results:Whereas the untreated diabetic rats showed a significant decrease(P<0.05)in packed cell volume,superoxide dismutase,catalase and high-density lipoprotein-cholesterol with a concomitant increase in the levels of malondialdehyde,fasting blood sugar,aspartate aminotransferase,alanine aminotransferase,bilirubin,urea and creatinine,administration of methanolic extract of T.tetraplera leaf or glibenclamide alleviated these altered parameters in the treated rats.Conclusions:Methanolic extract of T.tetraplera leaves possesses a potent capacity for treatment of diabetes and the accompanying complications,including oxidative stress and hyperlipidemia.

Involvement of mitochondrial swelling in cytochrome c release from mitochondria treated with calcium and Alloxan

An early event in the induction of apoptosis is cytochrome c (Cyt c) release from mitochondria. We investigated the involvement of mitochonrial permeability transition (MPT) and mitochondrial swelling in Cyt c release from mitochondria treated with alloxan and/or calcium (Ca2+). When mitochondria were treated with a high concentration of Ca2+. alone or Ca2+. with alloxan (alloxan-Ca2+), the MPT was accompanied by mitochondrial swelling and the release of Cyt c. Cyclosporin A prevented the induction of MPT but only slightly decreased the release of Cyt c. High molecular weight polyethylene glycol almost completely inhibited MPT-dependent osmotic mitochondrial swelling and Cyt c release. However, MPT-independent mitochondrial swelling and Cyt c release induced by exogenous K+. were inhibited by the high molecular weight polyethylene glycol. Ruthenium red strongly decreased the amount of Cyt c released. These results suggest that mitochondrial swelling but not MPT is necessary for Cyt c release induced by Ca2+ alone or alloxan and Ca2+.

Alloxan诱导制作糖尿病小鼠模型的影响因素研究

[目的]探讨Alloxan诱导制作糖尿病小鼠模型的影响因素.[方法]取昆明种雄性小鼠130只,随机选择10只作为对照组,取其余120只利用静脉注射和腹腔注射给予Alloxan方法制作糖尿病模型.静脉注射组分为3个组,分别静脉注射给予Alloxan 25,50,75mg/kg,腹腔注射组亦分为3个组,分别腹腔注射给予Alloxan 150,200,250mg/kg.记录制作模型的成功率和死亡率,观察第4,7d时的小鼠体质量和血糖值变化并进行比较.[结果]静脉注射组和腹腔注射组制作模型成功率平均分别为65%和80%,死亡率分别为35%和20%.实验第4d时,随着Alloxan剂量的增加,小鼠体质量和血糖水平均增高,实验第7d时,随着Alloxan剂量的增加,小鼠体质量降低而血糖水平增高,与对照组比较差异均有统计学意义(P<0.05).[结论]给药途径和给药剂量对Alloxan诱导制作糖尿病小鼠模型均有影响,腹腔注射给药途径制作模型的成功率较高,而Alloxan注射剂量在200mg/kg时制作的模型较稳定.

Effect of Doum Fruit (Hyphaene Thebaica) Extract on Some Biochemical Parameters, Enzyme Activities and Histopathological Changes of Pancreas in Alloxan Induced Diabetic Rats

Medicinal plants have been used in the treatment of some diseases like diabetes. One of them is doum fruit extract. The study was conducted to evaluate effect of doum fruit extract on some biochemical parameters, including enzyme activities and histopathological changes of pancreas in alloxan induced diabetic rats. Animals were allocated into five groups of rats, two groups, several as control (one normal and one diabetic rats) and the other three groups were induced diabetic with doum extract (0.5, 1 and 2 g/kg b&middot;wt) which were given every other day by stomach tube for 6 weeks. Evaluation was made for glucose, insulin, lipid profile, enzyme activities, antioxidant activity and histopathological changes of pancreas. The results showed that doum fruit extract decreased blood glucose, insulin, total cholesterol (TC), triglyceride (TG), total lipid (TL), low-density lipoprotein cholesterol (LDL-C), lactate dyhadreygenase (LDH), alanine and aspartate amino transferase (ALT, AST), alkaline phosphatase (ALP) and Malondialdehyde (MDA) while plasma catalase activity and high density lipoprotein cholesterol (HDL-C) were significantly (p 0.05) increased. Therefore, the results of this study demonstrated that the active compounds present in the doum extract have a positive effect on the biochemical parameters of diabetic rats.

Insulin Release from the Beta Cells in Acatalasemic Mice Is Highly Susceptible to Alloxan-Induced Oxidative Stress

Background: Catalase deficiency (acatalasemia) is sensitive to alloxan, and the administration to acatalasemic mice develops hyperglycemia under mild conditions. However, the mechanism is still poorly understood. Methods: Alloxan was used to induce the oxidative stress and intraperitoneally administered to acatalasemic and normal mice. The blood samples of these mice after 1, 3, 5 and 7 days were examined. The pancreatic islets 7 days after alloxan administration were isolated, and the insulin released under 3 mM and 20 mM glucose was examined. Results: After alloxan administration, increase of oxidative markers in blood and pancreatic apoptosis in acatalasemic mice were observed immediately. Insulin in blood was lowered after 3 days, and the insulin in acatalasemic mice was lower than that in normal mice. Hyperglycemia in the acatalasemic mice was observed after 3 days. The pancreatic islets after 7 days were isolated. A reduction of the insulin released from the islets under glucose stimulation was observed. The stimulation indexes of the normal and acatalasemic mice were 1.4 ± 0.6 and 0.7 ± 0.3, respectively. Conclusions: Alloxan induced a deterioration of glucose-dependent insulin secretion ability from the islets, and the deterioration mostly contributed to hyperglycemia, rather than apoptosis.

Hypoglycemic Effect Study of a Combination of Some Stipulated Spices in Alloxan Induced Diabetic Wistar Albino Rats along with Nutritional Value Evaluation

The main purpose of the study was to reveal the hypoglycemic effect of a combination of some selected spices along with the evaluation of nutritional values. A sum of fourteen different types of spices at different percentages were taken into consideration to propel the research activities forward, viz;ginger, garlic, onion, red chili, turmeric, cumin, cinnamon, clove, coriander, fenugreek, black pepper, nigella, carom seed and cardamom. The healthy experimental wistar albino rats were made diabetic by injecting alloxan monohydrate intraperitoneally a dose of 150 mg/kg body weight. The rats of Group-D and Group-E were treated with the sample, mixed spices at a dose of 100 mg/kg and 200 mg/kg body weight respectively with the lab diet while in case of Group-C, glibenclamide at a dose of 0.5 mg/kg body weight was introduced with the lab diet for a total period of 21 days where as for the same days span, Group-A and Group-B were continued feeding with only lab diet. Data suggest that the mixed spices of both doses applied in Group-D and Group-E had exerted effects significantly (p 0.05) in lowering the blood glucose level. On 22nd day, in Group-D and Group-E, the FBG level was found to be reduced by around 40.66% and 41.18% respectively compared to their respective initial days where in Group-C, the reduction was around 51.90% in comparison to its initial day. Again, in case of 2hPG level measurement, on 22nd day, both the groups, Group-D and Group-E were found with the decreased level of around 19.38% and 39.11% respectively compared to their respective initial days while the Group-C was shown with the reduced of 44.26% in comparison adopting same manner. Furthermore, various analyses confirm the presence of several micro and macro nutrients and high amount of dietary fiber in the spices mix. Thus the result affirms that this combined spice possesses strong hypoglycemic properties.