The chemical constituents and hypoglycemic activity of alcoholic extract of sea buckthorn leaves / 药学学报

The purpose of this research is to identify the chemical constituents of sea buckthorn leaves extract (SBLE) and explore its hypoglycemic biological activity. SBLE was prepared by hot reflux extraction with 65% ethanol, and its chemical composition was analyzed by ultra-high-performance liquid chromatography-photodiode array-mass spectrometry/mass spectrometry (UHPLC-PDA-MS/MS) system. The animal experiments were compliant with ethical principles for animal use and had been approved by the Animal Experiment Ethics Committee of Jinan University. Mice were injected with streptozocin (STZ) to establish a hyperglycemic animal model, and SBLE (1.5 g·kg-1) was administered by gavage for 5 weeks. The fasting blood glucose (FBG) and oral glucose tolerance were detected. Normal mice were given SBLE (1.5 g·kg-1) by intragastric administration for 10 days, and blood was collected from the tail vein to detect the changes in blood glucose within 120 min after sucrose or starch loading. The mucous membrane of the small intestine of mice was taken to detect the activity of α-glucosidase (AG), and the activity of yeast-derived AG incubated with SBLE was evaluated. The glucose uptake by Caco-2 cells treated with SBLE was detected by fluorescence microscopy and cytometry, and the gene expression of sodium-dependent glucose transporter 1 (SGLT1) and glucose transporter 2 (GLUT2) in Caco-2 cells were detected by real-time quantitative PCR (qPCR). A total of 18 compounds were identified, mainly including tannins and flavonoids. SBLE reduced FBG and increased oral glucose tolerance in STZ hyperglycemic mice. SBLE effectively inhibited the increase of blood glucose caused by starch intake in normal mice. SBLE exerted good inhibitory activity on yeast-derived AG (IC50 = 16.94 μg·mL-1) and small intestinal mucosa AG with an inhibition rate of 15.48%. SBLE (25-100 μg·mL-1) dose-dependently inhibited glucose uptake by Caco-2 cells, and SBLE significantly reduced the mRNA level of SGLT1 without changing the expression of GLUT2. In conclusion, the UHPLC characteristic fingerprint of SBLE is established with 18 chemical components identified by mass spectrometry, and SBLE exerts hypoglycemic effect by inhibiting the activity of AG and the absorption of glucose by intestinal epithelial cells.

Chronic treatment with hydroalcoholic extract of Plathymenia reticulata promotes islet hyperplasia and improves glycemic control in diabetic rats / Tratamento crônico com extrato hidroalcoólico de Plathymenia reticulata promove hiperplasia de ilhotas e controle glicêmico em ratos diabéticos

ABSTRACT Objective: To investigate the anti-hyperglycemic effects of Plathymenia reticulata hydroalcoholic extract and related changes in body weight, lipid profile and the pancreas. Methods: Diabetes was induced in 75 adult male Wistar rats via oral gavage of 65mg/Kg of streptozotocin. Rats were allocated to one of 8 groups, as follows: diabetic and control rats treated with water, diabetic and control rats treated with 100mg/kg or 200mg/kg of plant extract, and diabetic and control rats treated with glyburide. Treatment consisted of oral gavage for 30 days. Blood glucose levels and body weight were measured weekly. Animals were sacrificed and lipid profile and pancreatic tissue samples analyzed. Statistical analysis consisted of ANOVA, post-hoc Tukey-Kramer, paired Student's t and χ2 tests; the level of significance was set at 5%. Results: Extract gavage at 100mg/kg led to a decrease in blood glucose levels in diabetic rats in the second, third (198.71±65.27 versus 428.00±15.25) and fourth weeks (253.29±47.37 versus 443.22±42.72), body weight loss (13.22±5.70 versus 109.60±9.95) and lower cholesterol levels (58.75±3.13 versus 80.11±4.01) in control rats. Extract gavage at 200mg/Kg led to a decrease in glucose levels on the fourth week in diabetic rats, body weight loss in the second, third and fourth weeks in control rats, and lower cholesterol levels in diabetic and control rats. Islet hyperplasia (p=0.005) and pancreatic duct dilation (p=0.047) were observed in diabetic and control rats. Conclusion: Plathymenia extract reduced blood glucose levels in diabetic rats, and body weight in control rats, and promoted pancreatic islet hyperplasia in diabetic and control rats.

RESUMO Objetivo: Avaliar o efeito anti-hiperglicêmico do extrato hidroalcoólico de Plathymenia reticulata, alterações no peso, lipídeos e efeito sobre o pâncreas. Métodos: O diabetes foi induzido pela administração de estreptozotocina 65mg/kg, em 75 ratos Wistar adultos machos, divididos em 8 grupos diferentes: ratos diabéticos e controle + água, ratos diabéticos e controle + 100mg/kg ou 200mg/kg de extrato, ratos diabéticos e controle + gliburida. O tratamento foi realizado por gavagem (oral) por 30 dias. Níveis de glicose e peso foram verificados semanalmente. Os animais foram sacrificados, e amostras de lipídeos e do pâncreas foram analisadas. A análise estatística incluiu ANOVA, post-hoc Tukey-Kramer, teste t de Student pareado e teste do χ2, com nível de significância de 5%. Resultados: O extrato 100mg/kg promoveu redução nos níveis de glicose sanguínea em ratos diabéticos na segunda, terceira (198,71±65,27 versus 428,00±15,25) e quarta semanas (253,29±47,37 versus 443,22±42,72), perda de peso (13,22±5,70 versus 109,60±9,95) e diminuição do colesterol (58,75±3,13 versus 80,11±4,01) em ratos controle. Com extrato de 200mg/kg, houve redução dos níveis de glicose na quarta semana, nos ratos diabéticos; de peso na segunda, terceira e quarta semanas, nos ratos controle; e de colesterol nos animais diabéticos e controle. Ocorreram hiperplasia de ilhotas (p=0,005) e dilatação dos ductos pancreáticos (p=0,047) em ratos diabéticos e controles. Conclusão: O extrato de Plathymenia reduziu os níveis de glicose em ratos diabéticos e de peso em ratos controle, além de ter promovido hiperplasia de ilhotas pancreáticas em diabéticos e controles.

Pulmonary oxidative stress in diabetic rats exposed to hyperoxia

Abstract Purpose: To evaluate the pulmonary oxidative stress in diabetic rats exposed to hyperoxia for 90 minutes. Methods: Forty male Wistar rats were divided into four groups, each one containing 10 animals, according to the oxygen concentration to which they were exposed: 21%, 50%, 75% and 100% (hyperoxia). In each group five animals were randomly induced to diabetes by means of at a dose of 55 mg/kg of streptozotocin (STZ). Results: Seventy two hours after diabetes induction, a significant difference was seen in blood glucose in the experimental groups in comparison with the control. In the experimental groups a significant difference was observed in the concentration of malondialdehyde (MDA) in lung tissue and blood plasma (p<0.05), except the 50% group. In the control group, significant differences in the MDA concentration in plasma and lung tissue were also observed (p<0.05), except the 75% group. The MDA concentration in lung tissue in comparison with the diabetic and non-diabetic groups showed a significant difference in the 21% group; however, no difference was seen in the 75 and 100% groups. Conclusion: In diabetic animals high oxygen concentrations (75 and 100%) do not appear to exert deleterious effects on lipid peroxidation in lung tissue.

Study on Glycolipid Metablism of Mice with Diabetes Induced by Peptide Receptor Antagonist Pro3(GIP) / 现代检验医学杂志

Objective To investigate the metabolic effects of glucose dependent insulinotropic peptide receptor antagomst pro3 (GIP) in induced diabetes mice about blood glucose,triglyceride,cholesterol,leptin and fatty issue.Methods 27 C57 mice were randomly divided into normal group and diabetes mice group,and the mice in diabetes group were fed with high fat food and intraperitoneal injected streptozocin.Then 1 mouse that random blood giucose lower than 16.9 mmol/L was deleted in diabetes group.The rest mice in diabetes group were divided into two groups,diabetes control group,pro3 (GIP) group.Pro3 (GIP) group was given drug pro3 (GIP).The bloodglucose and glucose tolerance were measured.After treatment for 6 weeks,all mice were sacrificed and fatty tissues were collected.Results After 6 weeks,the blood glucose of the pro3 (GIP) group was obviously lower than diabetes control group (t=8.43,P＜0.01),and insulin levers in 0,30,60 and 120 min were obviously lower than diabetes control group (t =3.90,2.60,6.88 and 3.33,P＜0.05).There was significant difference between pro3 (GIP) group and diabetes control group about inflammatory cells.Moreover,leptin in pro3 (GIP) group was obviously lower than in diabetes control group (t =5.04,P＜0.01),but triglyceride,cholesterol,and adiponectin had no significant difference between two groups.Conclusion Pro3 (GIP) can significantly reduce blood glucose,insulin level,leptin of diabetes mice,and attenuate the inflammatory cells infiltration in fatty issue.

Therapeutical effect of mesenchymal stem cells overexpressing ICAM-1 in murine type 1 diabetes nephropathy / 军事医学

Objective To explore the curative effects of mesenchymal stem cells(MSC)that overexpress in murine type 1 diabetes nephropathy (DN).Methods Mice were randomly divided into normal control(NC) group,DN group,C3-treated group,C3-MIGR1-treated group and C3-MIGR1-ICAM-1-treated group.Mice were given streptozotocin until the DN model was set up.The murine DN model was treated with murine MSC(C3H10T1/2),transfection empty vector of murine MSCs(C3H10T1/2-MIGR1/MSC) and murine MSCs (C3H10T1/2-ICAM-1/MSC)that overexpressed ICAM-1.After transplantation, the pathological features of kidneys were observed by Masson staining and the number of homing MSC cells to the kidney was calculated on days 1,3,7 by frozen section, while qPCR was used to analyze the expression of signaling molecules for collagen1, TGF-β1 and SMAD2 after treatment with various MSCs.Results Compared with DN group, the renal fibrosis treated with MSCs overexpressing ICAM-1 was significantly decreased by Masson staining.Three and seven days after transplant, the homing cells of MSC in different groups displayed no difference using tissue freezing section method.Furthermore, TGF-β1/SMAD signaling was lowly activated after the treatment with MSCs that overexpressed ICAM-1 compared with model mice(P<0.01).Conclusion MSCs that overexpress ICAM-1 can protect kidneys in the DN model.

Determination of the critical diabetes duration in a streptozotocin-induced diabetic rat calvarial defect model for experimentation regarding bone regeneration

PURPOSE: The purpose of this study was to determine the critical diabetes duration in a streptozotocin (STZ)-induced diabetic rat calvarial defect model for experimentation regarding bone regeneration by evaluating the association between diabetes duration and bone healing capacity through histological and radiographic analyses. METHODS: Experimental diabetes was induced in 50 of 60 rats by an STZ injection. The rats were divided into 5 groups, including a control group (group 1), according to diabetes durations of 0, 2, 4, 6, and 8 weeks, respectively. Eighteen rats survived: 4 in group 1, 4 in group 2, 4 in group 3, 5 in group 4, and 1 in group 5. Calvarial defects were created at 0, 2, 4, 6, and 8 weeks after STZ injection in groups 1–5. Cone-beam computed tomography scanning was performed at baseline and at 5 and 7 weeks after surgery. The rats were sacrificed 7 weeks after surgery, followed by histological evaluation. RESULTS: The voxel gray values (VGVs) of group 1 and group 2 increased, whereas the VGVs of group 3 and group 4 decreased starting 5 weeks after surgery, although this trend did not reach statistical significance between groups. On the reconstructed 3-dimensional images and based on an analysis of histological features, groups 1 and 2 showed apparent bone regeneration, while groups 3–5 showed very limited bone regeneration. CONCLUSIONS: The critical diabetes duration in an STZ-induced diabetic rat calvarial defect model for experimentation regarding bone regeneration was between 2 and 4 weeks. It is suggested that researchers who use STZ-induced diabetic rats wait for more than 2 weeks following diabetes induction before placing implants or conducting bone regeneration studies to allow definite disturbances in bone healing to emerge.

Improvement of erectile dysfunction by the active pepide from urechis unicinctus by high temperature/pressure and ultra - wave assisted lysis in streptozotocin induced diabetic rats

ABSTRACT Introduction: We investigate the effect of active peptide from Urechis unicinctus (UU) by high temperature/pressure and ultra-wave assisted lysis on erectile dysfunction in streptozotocin-induced diabetic rats. Materials and Methods: Forty 12-week-old Sprague-Dawley rats were used in this study. Diabetes was induced by a one-time intraperitoneal injection of streptozotocin (50mg/kg). One week later, the diabetic rats were randomly divided into four groups: normal control, untreated diabetes control, and groups treated with 100 or 500mg/kg/d UU peptide. Rats were fed with UU peptide by intragastric administration for 8 weeks. After 8 weeks, penile hemodynamic function was evaluated in all groups by measuring the intracavernosal pressure after electrostimulating the cavernous nerve. Nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) activities were measured and endothelial nitric oxide synthase (eNOS) and neuronal NOS (nNOS) protein expression was determined by Western blot. Results: Maximum intracavernosal pressure in diabetic control rats decreased significantly compared to normal control rats, and was increased significantly compared to untreated diabetic rats after UU peptide supplementation. Treatment with the higher dose of UU peptide significantly increased the NO and cGMP levels compared with the diabetic control group. Decreased activity and expression eNOS and nNOS were found in the diabetic rats compared with the normal control group. Decreased eNOS and nNOS in diabetic rats were improved by UU peptide administration. Conclusions: Active peptide from UU ameliorates erectile function in a streptozotocin induced diabetic rat model of erectile dysfunction.

Endoplasmic reticulum stress and renal lesion in mice with combination of high-fat diet and streptozotocin-induced diabetes

ABSTRACT PURPOSE : To investigate in the kidney the pathologic changes and expression of GRP78 and CHOP in the Kunming (KM) mice with combination of high-fat diet and streptozotocin-induced diabetes. METHODS : Sixty two male KM mice were randomly divided into a normal control (NC) group (n=20) and a high-fat diet (HFD) group (n=42). After a four-week dietary manipulation, the KM mice in the HFD group were injected intraperitoneally with streptozotocin to induce diabetes. After diabetic models were successfully established, the kidneys were excised and conserved for further test. RESULTS : No significant difference in the body weight was observed after the dietary manipulation (p=0.554). After the streptozotocin was injected, fasting blood glucose levels in the diabetes group (DM) were significantly higher than that in the NC group (p＜0.0001). Glomerular atrophy observed under light microscope in the DM group was more serious compared with the NC group. The expression of GRP78 and CHOP in the kidneys of the mice in the DM group were higher compared with the NC group. CONCLUSION : Renal lesion occurs in the diabetic Kunming mice induced by combination of high-fat diet and low-dose streptozotocin, and endoplasmic reticulum stress and CHOP may contribute to the injury process.

Study on antidiabetic effect of aqueous extract from Pulsatilla chinensis / 中草药

Objective: To study the blood glucose lowering activity of aqueous extract from Pulsatilla chinensis (AEPC) on Streptozocin (STZ)-induced type-2 diabetic rats. Methods: AEPC was obtained through traditional aqueous extraction procedure; Oral glucose tolerance test was employed for the preliminary study of AEPC's blood glucose lowering activity; For further study, STZ-induced neonatal diabetic rat model was used, and the serum SGPT, SGOT, ALP, serum urea, triglycerides, total cholesterol, HDLC, total protein, and serum insulin were tested; The Kunming rats were used for acute toxicity experiments, with the highest dose of 1 000 mg/kg body weight; Insulin release from RIN 5F cells was performed to study the mechanism of AEPC on antidiabetes effect. Results: Oral glucose tolerance test showed AEPC could lower the blood sugar, and the optimal activity was observed at a dose of 20 mg/kg body weight; In neonatal STZ-induced type-2 diabetic rats, treatement with 20 mg/kg AEPC for 20 d resulted in significant blood glucose lowering activity, liver glycogen and serum biological parameters back to normal; The acute toxicity results showed that no obvious toxicity was observed even at 1 000 mg/kg dose; At last, AEPC could promote the secretion of insulin from RIN 5F cells. Conclusion: AEPC shows the anti-diabetes activity on STZ-induced type-2 diabetic rats, which may be the result of its effect on promoting insulin release from pancreatic α-cells.

Effect of maternal diabetes on female offspring / Efeito do diabetes materno na prole feminina

Objective To evaluate the effect of maternal diabetes on the blood pressure and kidney function of female offspring, as well as if such changes exacerbate during pregnancy. Methods Diabetes mellitus was induced in female rats with the administration of streptozotocin in a single dose, one week before mating. During pregnancy, blood pressure was measured through plethysmography. On the 20th day of pregnancy, the animals were placed for 24 hours in metabolic cages to obtain urine samples. After the animals were removed from the cages, blood samples were withdrawn. One month after pregnancy, new blood and urine sample were collected. Kidney function was evaluated through proteinuria, plasma urea, plasma creatinine, creatinine excretion rate, urinary flow, and creatinine clearance. Results The female offspring from diabetic mothers showed an increase in blood pressure, and a decrease in glomerular filtration rate in relation to the control group. Conclusion Hyperglycemia during pregnancy was capable of causing an increase in blood pressure and kidney dysfunction in the female offspring. .

Objetivo Avaliar o efeito do diabetes materno sobre a pressão arterial e a função renal da prole feminina, bem como verificar se as alterações observadas se exacerbam durante a prenhez. Métodos O diabetes mellitus foi induzido em ratas com a administração de estreptozocina em dose única, uma semana antes do cruzamento. Durante a prenhez, foram feitas medidas da pressão arterial por pletismografia. No 20o dia da prenhez, os animais foram colocados durante 24 horas em gaiolas metabólicas para obtenção de amostras de urina. Após a retirada dos animais das gaiolas, foram obtidas amostras de sangue. Um mês após a prenhez, foram obtidas novas amostras de sangue e urina para as determinações. A função renal foi avaliada por meio de proteinúria, ureia plasmática, creatinina plasmática, carga excretada de creatinina, fluxo urinário e clearance de creatinina. Resultados As fêmeas da prole de mães diabéticas apresentaram elevação da pressão arterial e redução do ritmo de filtração glomerular em relação ao grupo controle. Conclusão A hiperglicemia durante a gestação foi capaz de causar elevação da pressão arterial e disfunção renal na prole de sexo feminino. .

MAPK immunoreactivity in streptozotocin-induced diabetic rat testis

PURPOSE: To evaluate the alterations of two mitogen-activated protein kinases (MAPK)s, extracellular signal regulated kinase (ERK) and c-Jun NH2 terminal kinase (JNK), in the testes of male rats with experimental diabetes. METHODS: Twenty males Sprague-Dawley rats were randomly divided into a control group (n=8) and a diabetes group (administration of 40 mg/kg/day streptozotocin (STZ) for five sequential days, n=12). After six weeks, testicular biopsy samples were obtained for light microscopy and immunohistochemical methods. RESULTS: The PCNA (proliferating cell nuclear antigen) index was significantly decreased in the diabetes group (p=0.004) when compared to the control group. Both total (t)-ERK and phosphor (p)-ERK immunoreactivities were significantly decreased in the diabetes group (p=0.004, p<0.001, respectively). The t-JNK immunoreactivity was unchanged in both groups (p=0.125), while p-JNK immunoreactivity was significantly increased in the diabetic group (p=0.002). CONCLUSIONS: The decrease of androgen levels in the course of diabetes may contribute to the decrease of the immunoreactivities of t-ERK and p-ERK. JNK may be activated due to the changes in various cytokines and chemochines that participate in the oxidative stress process of diabetes. Therefore, testicular apoptosis may occur and lead to infertility associated with diabetes. .

Effects of the Diabetic Condition on Grafted Fat Survival: An Experimental Study Using Streptozotocin-Induced Diabetic Rats

BACKGROUND: Autologous fat grafts have been widely used for cosmetic purposes and for soft tissue contour reconstruction. Because diabetes mellitus is one of the major chronic diseases in nearly every country, the requirement for fat grafts in diabetes patients is expected to increase continuously. However, the circulation complications of diabetes are serious and have been shown to involve microvascular problems, impairing ischemia-driven neovascularization in particular. After injection, revascularization is vital to the survival of the grafted fat. In this study, the authors attempted to determine whether the diabetic condition inhibits the survival of injected fat due to impaired neovascularization. METHODS: The rat scalp was used for testing fat graft survival. Forty-four seven-week-old male Sprague-Dawley rats were allocated to a diabetic group or a control group. 1.0 mL of processed fat was injected subcutaneously into the scalp of each rat. The effect of diabetes was evaluated by calculating the volume and the weight of the grafted fat and by histologically analyzing the fat sections. RESULTS: The surviving fat graft volume and weight were considerably smaller in the diabetic group than in the control group (P<0.05), and histological evaluations showed less vascularity, and more cysts, vacuoles, and fibrosis in the diabetic group (P<0.05). Cellular integrity and inflammation were not considerably different in the two groups. CONCLUSIONS: As the final outcome, we found that the presence of diabetes might impair the survival and the quality of fat grafts, as evidenced by lower fat graft weights and volumes and poor histologic graft quality.

Study of the method used to induce the model of type 2 diabetic mellitus rat and the characteristic of the nephropathy / 中国医师杂志

ObjectiveTo discuss the feasibility of the method that can be used to induce the model of type 2 diabetic mellitus rat and explore the characteristic of the nephropathy.The rats were fed with the high sucrose,high fat and high energy feed for a long time and then it was injected with the low dose STZ.Methods30 SD rats were selected and then it was randomly divided them into 2 groups,the control group (10 rats) and the model group (20 rats).The model group was fed with the high calorie feed forl0 weeks to induce insulin resistance and then the rats were induced to type 2 diabetes mellitus by injection of streptozocin (30 mg/kg).The rats of the model group were continually fed with the high calorie feed for 2 months.Before the end of this study,the 24-hours microalbuminuria,serum creatinine and blood urea nitrogen were detected and the periodic acid Schiff staining on the kidney were also measured.ResultsAfter the rats of the model group were established,the levels of the bodyweight,the cholesterol,the triglyceride and the insulin were [ (468.7 ± 8.8 ) g,( 1.92 ± 0.27 ) mmol/L,( 1.32 ± 0.34) mmol/L,(38.81 ± 5.39 ) mU/L ] respectively,all of them were higher than the levels in the normal group,which were [ (436.9 ± 7.4) g,(1.16 ±0.17)mmol/L,(0.8 ±0.18)mmo1/L,(21.43 ±4.19)mU/L],respectively( t =9.755,8.077,4.437,8.902,P ＜ 0.01 ).After injection of STZ for 2 weeks,the levels of the blood glucose,the insulin and the insulin resistance of the diabetes mellitus rats were [ ( 19.31 ± 1.55 ) mmol/L,( 31.31 ± 8.60) mU/L,(26.55 ± 6.33) ] respectively,it was higher than levels inf the normal group,which was[ (5.45 ±0.69) mmol/L,( 19.97 ± 3.26) mU/L,(4.82 ± 0.84) ] ( t =26.383,3.951,10.719,P ＜ 0.01 ).Before the end of this study,the levels of the blood glucose,insulin resistance,24-hours microalbuminuria,serum creatinine,blood urea nitrogen of the diabetes mellitus rats were [ ( 19.27 ± 1.97 ) mmol/L,( 16.70 ±7.51 ),(72.49 ± 8.53 ) mg/24 h,( 74.76 ± 8.38 ) μmol/L,( 19.09 ± 4.21 ) mmol/L],it was higher than the levels in the normal group,which were [ (5.62 ±0.65) mmol/L,(5.45 ± 1、33),( 15.26 ±2.20) mg/24 h,(40.81 ± 1.97) μmol/L,(9.87 ±2.13) mmol/L,t =20.961,4.657,20.623,12.495,6.352,P ＜0.01 ].And the pathological changes of the diabetes mellitus rats kidney tissues were the most serious through themethod of periodic acid-Schiff's staining (PAS).ConclusionsThe model of type 2 diabetic mellitus rat was constructed through the way of feeding the SD rats with high sucrose,high fat and high energy feed for a long time and low dose STZ.The diabetic mellitus rats had the symptom of drinking more,eating more and diuresis,and the character of this model had high levels of albuminuria,serum creatinine,blood urea nitrogen.The incrassated glomerular mesangium,the crescent-shaped focus and the glomerulosclerosis were also observed through the PAS.

Análise da presença de citocinas no periodonto de ratos diabéticos tratados com chá verde / Cytokines expression analyses in periodontium of diabetic rats treated with green tea

As doenças periodontais (DPs) são alterações inflamatórias crônicas que acometem os tecidos de sustentação do órgão dental. A presença do diabetes é refletida em maior severidade e prevalência das DPs tanto em humanos quanto em modelos experimentais. Contudo, os mecanismos biológicos envolvidos no aumento da prevalência e da severidade permanecem pouco conhecidos. Desta forma, o objetivo deste estudo foi avaliar o número de células marcadas por imunohistoquímica para TNF-α, RANKL, OPG, IL-10 e para o fator de transcrição RUNX-2, na doença periodontal experimental decorrente da indução do diabetes em ratos. Além disso, avaliamos os possíveis efeitos do tratamento com chá verde sobre o periodonto dos animais. Inicialmente, os ratos (n=80) foram submetidos à indução do diabetes por administração intraperitoneal de estreptozotocina (50mg/kg) e, juntamente com o grupo controle (n=40), foram subdivididos em animais tratados com chá verde ou com água, acompanhados durante o período de 15, 30, 60 ou 90 dias. Após o sacrifício dentro do tempo determinado para cada grupo, as hemimaxilas coletadas passaram pelos procedimentos de imunohistoquímica. Os resultados revelaram que a presença do diabetes causou perda óssea alveolar, compatível com o desenvolvimento da doença periodontal e resultou em alterações significativas no número de células imunomarcadas para diferentes mediadores do processo inflamatório. Entretanto, o chá verde apresentou efeitos benéficos para o periodonto, alterando a marcação das citocinas envolvidas. Nos animais diabéticos, independente do tratamento, foi observado aumento estatisticamente significativo (p<0,05 ANOVA) no número de células imunomarcadas para TNF-α e RANKL. Inversamente, houve menor marcação para OPG (60 e 90 dias), RUNX-2 (30, 60 e 90 dias) e para IL-10 (30, 60 e 90 dias) nos animais que ingeriram água. Porém, os diabéticos tratados com chá não demonstraram diferenças significativas em relação ao seu respectivo controle...

Periodontal diseases (PD) are chronic inflammatory diseases leading the destruction of connective tissue and alveolar bone supporting the teeth. The establishment of diabetes increases PD prevalence and severity in humans and experimental model. However, biological mechanisms regarding to increase of prevalence and severity remains poorly known. The aim of this study was to evaluate the number of immuno-staining cells to TNF-α, RANKL, OPG, IL-10 and transcription factor RUNX-2 in experimental periodontal disease in diabetic rats. Furthermore, the possible green tea efects were evaluated in periodontiumof the rats. Diabetes was induced in Wistar rats (n=120) by intraperitoneal administration of 50 mg/kg ofstreptozotocin and together with control animals (n=80), the rats were subdivided in water or green tea treated group, that were analyzed at 15, 30, 60 and 90 days after diabetes induction. The animals were sacrificed and the hemimaxillae were removed and submitted to immunohistochemistry procedures. Our data demonstrated that diabetes induction and progression resulted in significant bone loss and alterations in number of immuno-staining cells to different mediators of inflammatory process. However, the green tea showed positive effects in periodontium through inflammation modulation. In diabetic rats, regardless of treatment, we observed an increased number of immuno-staining cells to TNF-α, IL-1b and RANKL (p<0,05 ANOVA). On the other hand, in water treated diabetic rats, there were a decreased number of immuno-staining cells to OPG (60 e 90 days), RUNX-2 (30, 60 e 90 days) and IL-10 (30, 60 e 90 days). However, the green tea treated rats did not showed statistical differences between control and experimental groups in those staining. When we compared both diabetic groups, green tea and water treated, the animals that drank the green tea showed decreased number of immuno-staining cells to TNF-α and RANKL(p<0,05 ANOVA) whereas the number...

Análise da presença de citocinas no periodonto de ratos diabéticos tratados com chá verde / Cytokines expression analyses in periodontium of diabetic rats treated with green tea

As doenças periodontais (DPs) são alterações inflamatórias crônicas que acometem os tecidos de sustentação do órgão dental. A presença do diabetes é refletida em maior severidade e prevalência das DPs tanto em humanos quanto em modelos experimentais. Contudo, os mecanismos biológicos envolvidos no aumento da prevalência e da severidade permanecem pouco conhecidos. Desta forma, o objetivo deste estudo foi avaliar o número de células marcadas por imunohistoquímica para TNF-α, RANKL, OPG, IL-10 e para o fator de transcrição RUNX-2, na doença periodontal experimental decorrente da indução do diabetes em ratos. Além disso, avaliamos os possíveis efeitos do tratamento com chá verde sobre o periodonto dos animais. Inicialmente, os ratos (n=80) foram submetidos à indução do diabetes por administração intraperitoneal de estreptozotocina (50mg/kg) e, juntamente com o grupo controle (n=40), foram subdivididos em animais tratados com chá verde ou com água, acompanhados durante o período de 15, 30, 60 ou 90 dias. Após o sacrifício dentro do tempo determinado para cada grupo, as hemimaxilas coletadas passaram pelos procedimentos de imunohistoquímica. Os resultados revelaram que a presença do diabetes causou perda óssea alveolar, compatível com o desenvolvimento da doença periodontal e resultou em alterações significativas no número de células imunomarcadas para diferentes mediadores do processo inflamatório. Entretanto, o chá verde apresentou efeitos benéficos para o periodonto, alterando a marcação das citocinas envolvidas. Nos animais diabéticos, independente do tratamento, foi observado aumento estatisticamente significativo (p<0,05 ANOVA) no número de células imunomarcadas para TNF-α e RANKL. Inversamente, houve menor marcação para OPG (60 e 90 dias), RUNX-2 (30, 60 e 90 dias) e para IL-10 (30, 60 e 90 dias) nos animais que ingeriram água. Porém, os diabéticos tratados com chá não demonstraram diferenças significativas em relação ao seu respectivo controle...

Periodontal diseases (PD) are chronic inflammatory diseases leading the destruction of connective tissue and alveolar bone supporting the teeth. The establishment of diabetes increases PD prevalence and severity in humans and experimental model. However, biological mechanisms regarding to increase of prevalence and severity remains poorly known. The aim of this study was to evaluate the number of immuno-staining cells to TNF-α, RANKL, OPG, IL-10 and transcription factor RUNX-2 in experimental periodontal disease in diabetic rats. Furthermore, the possible green tea efects were evaluated in periodontiumof the rats. Diabetes was induced in Wistar rats (n=120) by intraperitoneal administration of 50 mg/kg ofstreptozotocin and together with control animals (n=80), the rats were subdivided in water or green tea treated group, that were analyzed at 15, 30, 60 and 90 days after diabetes induction. The animals were sacrificed and the hemimaxillae were removed and submitted to immunohistochemistry procedures. Our data demonstrated that diabetes induction and progression resulted in significant bone loss and alterations in number of immuno-staining cells to different mediators of inflammatory process. However, the green tea showed positive effects in periodontium through inflammation modulation. In diabetic rats, regardless of treatment, we observed an increased number of immuno-staining cells to TNF-α, IL-1b and RANKL (p<0,05 ANOVA). On the other hand, in water treated diabetic rats, there were a decreased number of immuno-staining cells to OPG (60 e 90 days), RUNX-2 (30, 60 e 90 days) and IL-10 (30, 60 e 90 days). However, the green tea treated rats did not showed statistical differences between control and experimental groups in those staining. When we compared both diabetic groups, green tea and water treated, the animals that drank the green tea showed decreased number of immuno-staining cells to TNF-α and RANKL(p<0,05 ANOVA) whereas the number...

Evaluation of placental glycogen storage in mild diabetic rats / Avaliação dos estoques de glicogênio placentário em ratas com diabete moderado

Purpose: To evaluate the placental glycogen storage and fetal development in the pregnancy of neonatally streptozocin-induced diabetic rats and to establish relation with glycemia and insulin levels. Methods: At the birth day, 147 female rats were randomly distributed in two experimental groups: 1) Non-diabetic Group (Control, n=45) - received the vehicle; 2) Diabetic Group (STZ, n=102) - received 100 mg streptozocin/kg in neonatal period. At day 0 of pregnancy, adult female rats were included in the control group when presented glycemia below 120 mg/dL and, in the group STZ with glycemia between 120 and 300 mg/dL. At day 21 of pregnancy, blood samples were collected for glycemia and insulin determination, and placentas withdrawn for placental glycogen determination. The newborns (NB) were classified in small (SGA), appropriate (AGA) and large (LGA) for gestational age. Results: Rats STZ presented higher glycemia at days 0 and 14 of pregnancy. At end of pregnancy, rats STZ showed higher proportion of NB SGA and LGA; reduced rate of NB AGA and unaltered glycemia, insulin and placental glycogen determinations. Conclusion: Mild diabetes altered the maternal glycemia in the early pregnancy, impairing future fetal development, but it caused no alteration on insulin and placental glycogen determination, confirming that this glycemic intensity was insufficient to change glycogen metabolism.

Objetivo: Avaliar os estoques de glicogênio placentário e o desenvolvimento fetal na prenhez de ratas com diabete moderado induzido no período neonatal e relacionar com glicemia e níveis de insulina. Métodos: No dia de nascimento, foram distribuídas aleatoriamente 147 ratas em dois grupos experimentais: 1) Grupo Não-diabético (Controle, n=45) - recebeu o veículo; 2) Grupo Diabético (STZ, n=102) - recebeu 100 mg streptozocin/kg peso corpóreo. No dia 0 de prenhez, foram incluídas ratas controle que apresentassem glicemia baixo de 120 mg/dL e, no grupo STZ, com glicemia entre 120 e 300 mg/dL. No 21º dia de prenhez, amostras de sangue foram coletadas para glicemia e determinação de insulina e as placentas foram retiradas para determinação de glicogênio placentário. Os recém-nascidos (RN) foram classificados em pequeno (PIP), adequado (AIP) e grande (GIP) para idade de prenhez. Resultados: As ratas STZ apresentaram glicemias maiores nos dias 0 e 14 de prenhez. No final da prenhez, as ratas STZ mostraram maior proporção de RN PIP e GIP, taxa reduzida de RN AIP e inalteração em glicemia, insulina e na determinação de glicogênio placentário. Conclusão: O diabete moderado alterou a glicemia materna no início da prenhez, prejudicando o futuro desenvolvimento placentário e fetal, mas não causou nenhuma alteração na determinação de insulina e de glicogênio placentário, confirmando que esta intensidade de glicêmica foi insuficiente para modificar o metabolismo de glicogênio.

Protective effect of Achyranthes bidentata polysaccharides on diabetic mice induced by streptozocin / 中国药理学与毒理学杂志

OBJECTIVE To investigate the potential protective effect of Achyranthes bidentata polysaccharides (ABP) on diabetes mice induced by streptozocin. METHODS Male ICR mice were divided into normal control, diabetes model and ABP 50 and 100 mg·kg~(-1) (ip, once daily for 15 d) treatment groups. On the day before ABP administration and after ABP administration for 8 and 15 d, the blood glucose content was detected with a glucometer and intraperitoneal glucose tolerance test was also conducted. After ABP administration for 15 d, the mice were sacrificed and body weight, heart, liver, spleen and kidneys weights were measured. The serum insulin concentration was determined by radioimmunoassay kit. The serum activities of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and alkaline phosphatase (ALP), and serum concentrations of triglyceride (TG), total cholesterol (TC), calcium and phosphorus were measured by colorimetric method. Enzyme linked immunosorbent assay was employed to detect the concentrations of leptin, adiponectin, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum. RESULTS Compared with mice in normal control group, the body weight and serum insulin concentration decreased and blood glucose increased in diabetic model mice. ABP 50 and 100 mg·kg~(-1) treated mice were able to normalize glucose concentrations better following a glucose tolerance test, and the blood glucose level decreased by 27.4% and 16.3%, respectively, compared with that of diabetic model mice. The relative weights of spleen and kidneys, blood glucose level, serum TG and TC concentrations, and GOT, GPT and ALP activities in mice treated with ABP 50 mg·kg~(-1) were obviously lower than those of diabetes model mice. Serum leptin concentration was also markedly decreased near to normal level. However, serum concentrations of adiponectin, TNF-α and IL-6 were significantly increased comparing with diabetes model mice. ABP 100 mg·kg~(-1) had no obvious effect on serum TG and TC levels, and GPT and ALP activities. Its effects on the other parameters indicated above were similar to those in ABP 50 mg·kg~(-1) group. For the serum concentrations of insulin, calcium and phosphorus, no statistical difference could be observed among the different groups. CONCLUSION ABP possesses protective effect against streptozocin-induced diabetes in mice.

Influence of streptozocin's dose on inductive effect of diabetes in C57BL/6J mice / 中华胰腺病杂志

Objective To investigate the influence of streptozocin (STZ)'s dose on the inductive effect of diabetes in C57BL/6J mice, and investigate the dose-effect relationship and the optimal dose range. Methods 145 C57BL/6J mice were randomly divided into 9 diabetic groups (group A to group 1, n = 15 in each group) and I control group (n = I0) to receive intraperitoneal injection of STZ with the dosages of 30, 60, 80, 100, 120, 150, 180, 210, 240 mg/kg and same amount of buffer solution,respectively. Changes of blood glucose, body weight, survival rate at 45 day and serum insulin level were monitored, and the relationship with STZ doses was analyzed. Pancreas and kidneys of the mice were removed for morphological examination, and immunohistochemistry was used for determination of insulin in pancreas and CD<,68> in kidneys. Results Compared with control group, blood glucose in group C ～G increased significantly; body weigh, insulin level decreased significantly (P < 0.05), and the STZ dose was positively correlated with mean blood glucose (r = -0.984, P < 0.05) and was negatively correlated with mean serum insulin levels (r = 0.994, P <0.05). The diabetes modeling rates in group C ～ G (86.7% ～ 100%) were higher than those of group A and B (0 and 40%, P<0.05). At the 45th day, the survival rates of group C ～G (46.7% ～ 73.3%) were higher than those of group H and 1 (13.3% and 0, P <0.05). There was no obvious injury of pancreas and kidneys in group B, whereas, in group C and G, pancreatic island atrophy and decreased insulin secretion were observed; deposits of extracellular matrix and macrophage increased in the mesangium were also present. Conclusions 80 ～ 180 mg/kg of STZ dose was ideal for establishing diabetes model in C57BL/6J mice. Within this range, the modeling rate and survival rate was higher, and target organs injury was typical. The STZ dose was positively correlated with blood glucose and negatively correlated with serum insulin levels.

Neonatally-induced diabetes: lipid profile outcomes and oxidative stress status in adult rats / Diabete induzido no período neonatal: repercussões no perfil lipídico e avaliação dos marcadores de estresse oxidativo na vida adulta de ratas

BACKGROUND: Experimental models are developed for the purpose of enhancing the understanding of the pathophysiological mechanisms involved in diabetes. Experimental findings lead to the development of treatment strategies to maintain metabolic conditions as close to normal as possible. There are several reports about streptozotocin induced mild diabetes to reproduce type 2 diabetes. However, studies about the interaction among glucose levels, lipid profile, and oxidative stress in these animals remain insufficient. Therefore, this study evaluated these parameters in blood samples from adult Wistar rats treated neonatally with streptozotocin. METHODS: Female newborn Wistar rats received streptozotocin (70 mg/kg, i.p.) on the 5th day of life (n5-STZ). Glycemia was measured in the 3rd and 4th month of life. At the end of the 4th month, blood samples were collected and processed for lipid profile and oxidative stress measurements. RESULTS: Glycemia of n5-STZ rats were significantly higher compared to those of control rats (p<0.05). There was no alteration in levels of total cholesterol, triglycerides, lipid peroxidation (TBARS), SOD activity and GSH-t determination (p>0.05) in the n5-STZ animals when compared to control group. However n5-STZ animals showed a significant decreased HDL-cholesterol rate (p<0.05). CONCLUSION: This streptozotocin-induced diabetes model in rats caused hyperglycemia (120-360mg/dL), characterizing mild diabetes. This glycemic level led to HDL-lipoprotein alteration, which was not sufficient to impair antioxidant enzyme activities or determination of lipid peroxidation in adult life of rats. Further this experimental investigation contributed to the understanding of different results found in other models for mild/moderate diabetes induction in laboratory animals as well as to a better comprehension of the pathophysiological mechanisms of mild diabetes or hyperglycemia in humans.

INTRODUÇÃO: Modelos experimentais são desenvolvidos com propósito de ampliar o entendimento dos mecanismos fisiopatológicos envolvidos no diabete. Os achados experimentais levam ao desenvolvimento de tratamentos alternativos para a manutenção das condições metabólicas normais. Existem vários estudos sobre o diabete induzido por streptozotocin mimetizando o quadro clínico do DM2. No entanto, a interação entre os níveis de glicose, perfil lipídico e estresse oxidativo nestes animais são escassos. Portanto, o objetivo do trabalho foi avaliar estes parâmetros em ratas Wistar adultas com diabete induzido com streptozotocin no período neonatal. MÉTODOS: Fêmeas recém-nascidas receberam streptozotocin (70mg/Kg, ip) no 5º dia de vida (n5-STZ). A glicemia foi medida no terceiro e quarto meses de vida dos animais. No final do quarto mês de vida, amostras de sangue foram coletadas e processadas para a dosagem de lipídios e marcadores de estresse oxidativo. RESULTADOS: A glicemia das ratas do grupo n5-STZ foi significativamente maior comparada às ratas do grupo controle (p<0,05). Não houve alteração nos níveis de colesterol total e triglicérides, peroxidação lipídica (TBARS), atividade da SOD e determinação da GSH-t (p>0,05) nas ratas n5-STZ em relação às ratas do grupo controle. No entanto, houve diminuição significativa no HDL-colesterol (p<0,05). CONCLUSÃO: Este modelo de indução de diabete em ratas causou hiperglicemia (120-360mg/dL), caracterizando o diabete moderado. Essa glicemia levou a alterações no HDL-colesterol, a qual não foi suficiente para prejudicar a atividade das enzimas antioxidantes ou marcadores da peroxidação lipídica na vida adulta. Além disso, esta investigação experimental contribuiu para entender os diferentes resultados encontrados em outros modelos deindução do diabete moderado em animais de laboratório, como também para a melhor compreensão dos mecanismos fisiopatológicos do diabete moderado ou da hiperglicemia em humanos.

Effects of Ketangte 2 on animal model of hyperglycemia / 中成药

AIM:To study the effect of Ketangte2 (KTT2)on reducing blood glucose and its mechanism.METHODS:Adrenalin and streptozocin were used to induce hyperglycemia in two mice models,and then treated them with glybenzcyclamide (50 mg/kg),three different dose of KTT2 and normal saline(NS)(0.1 ml/10 g weight)for 15 days.And other healthy mice were set up as control group,During the experiment,the fast blood glucose(FBG),insulin was measured in different time.The pancreas and islets of diabetic mice induced by STZ were studied by pathologic section.RESULTS:In the adrenalin-induced model group,KIT2 could lighten hyperglycemic reaction,which Was significant different from that of model group(P＜0.05 or 0.01),while KTT2 high,middle dose group could decrease blood glucose in streptozocin-induced hypoglycemic mice obviously(P＜0.01).Histological examination showed that pancreatic island number in pancreas in KTT2 groups increased in comparison with the model group.CONCLUSION:KTT2 can obviously reduce the blood glucose in the two kinds of animal models of hyperglycemia (P＜0.05 or 0.01).Its mechanism may be related to the protective effect on islet β cells,thereby increase the insulin secretion.