Free interstitial levels of metformin in the liver of healthy and diabetic Wistar rats

Abstract In the present study, free interstitial levels reached by metformin in the liver were investigated in control and diabetic rats by microdialysis. Firstly, a bioanalytical method using an HPLC-UV system to determine the drug concentration in microdialysis samples was validated. The blood glucose levels and biochemical parameters were investigated in control and diabetic animals. Following that, both groups received a dose of 50 mg/kg of metformin iv bolus and the free interstitial levels reached in the liver were assessed by microdialysis. The method was validated according to FDA guidelines being suitable to quantify free concentrations of metformin in the liver of control and diabetics rats. Free exposure to metformin was similar in control and diabetic animals: AUC0-∞ 118.50 ± 40.18 vs 112.93 ± 50.25 µg.h/mL, respectively. The half-life in tissue was similar to that described in the literature for plasma. Hence diabetes induced by streptozotocin after administration of nicotinamide in our study did not damage the renal and hepatic function of the animals. The levels reached in the liver were 1.6 times higher than the free plasma concentrations, demonstrating higher liver penetration of metformin. This is the first investigation in liver interstitial concentration of metformin in control and diabetic rats

Determination of the optimal diabetes duration for bone regeneration experiments in an alloxan-induced diabetic rabbit calvarial defect model

PURPOSE: The purpose of this study was to evaluate the optimal diabetes duration for bone regeneration experiments in an alloxan monohydrate (ALX)–induced diabetic rabbit calvarial defect model by evaluating the association between diabetes duration and bone healing capacity. METHODS: Twenty-four New Zealand white rabbits were used. Twenty-two rabbits were injected with 100 mg/kg of ALX to induce experimental diabetes. These rabbits were divided into 4 groups, including a control group and groups with diabetes durations of 1 week (group 1), 2 weeks (group 2), and 4 weeks (group 3). Calvarial defects were created at 1, 2, and 4 weeks after ALX injection and in the control rabbits. Cone-beam computed tomography (CBCT) scanning was performed on the day of surgery and at 2 and 4 weeks after surgery. The rabbits were sacrificed 4 weeks after surgery, followed by histological and immunofluorescence analysis. RESULTS: The diabetic state of all diabetic rabbits was well-maintained throughout the experiment. Reconstructed 3-dimensional CBCT imaging showed more rapid and prominent bone regeneration in the control group than in the experimental groups. Histological staining showed notable bone regeneration in the control group, in contrast to scarce bone formation in the experimental groups. The appearance and immunoreactivity of receptor activator of nuclear factor-kappa B and osteoprotegerin did not show notable differences among the groups. CONCLUSION: ALX administration at 100 mg/kg successfully induced experimental diabetes in rabbits. The effect of diabetes on bone healing was evident when the interval between diabetes induction and the intervention was ≥1 week.

Changes in hepatic metabolic enzyme activities and biliary excretion of 4-nitrophenol in streptozotocin induced diabetic rats

Abstract Activity of hepatic metabolic enzymes of glucuronidation and sulfation of 4-nitrophenol (PNP) and biliary excretion of its glucuronide (PNP-G) and sulfate (PNP-S) conjugates have been investigated in control and streptozotocin (STZ)-induced diabetic rats. 500 µM PNP solution was luminally perfused in a cannulated jejunal loop for 90 minutes. It was found that biliary excretion of PNP-G was significantly decreased in the diabetic rats. This effect of STZ could be completely reversed by administration of rapid-acting insulin. Activity of hepatic UDP-glucuronyltransferase and ß-glucuronidase was also depressed by the STZ pretreatment. Administration of insulin antagonized the inhibitory action of STZ on UDP-glucuronyltransferase, but the reduced activity of ß-glucuronidase was not reversed. Biliary excretion of PNP-S was also depressed in the diabetic rats. Whereas, different effects of insulin administration were observed. Namely, the lower biliary excretion rate of PNP-S was not changed after administration of insulin. Activity of the sulfotransferase and the arylsulfatase enzymes was not altered either by STZ pretreatment or by insulin administration. Biliary excretion of PNP was also significantly depressed by STZ and this depression was not changed after insulin administration. The results call attention to hepatobiliary circulation of low molecular weight xenobiotics and their glucuronide and sulfate conjugates

Avaliação das citocinas TNF-α, RANKL e OPG e do número de osteoclastos no reparo de defeito ósseo em calvária de ratos diabéticos tratados com matriz óssea desmineralizada / Evaluation of cytokines TNF-α, RANKL and OPG and the number of osteoclasts on repair of bone defects in skulls of diabetic rats treated with demineralized bone matrix

Neste trabalho, foi avaliado a participação dos osteoclastos bem como a ação das citocinas RANKL, OPG e TNF-α durante a formação e remodelação óssea em defeitos ósseos de tamanho crítico em ratos normoglicêmicos e diabéticos tratados ou não com a MAOD. Para isso, foram utilizados 250 ratos machos Wistar. Trinta ratos foram utilizados para coleta dos fêmures e tíbias, os quais foram processados para obtenção da MAOD. Os demais 220 ratos foram divididos em Grupo Não Diabétido (CTL, n=110) e Grupo Diabético (DIAB, n= 110) induzido pela aplicação de uma dose única de 47 mg/Kg de massa corporal de estreptozotocina. Um defeito transósseo de 8 mm de diâmetro foi realizado nos ossos parietais dos ratos, sendo que, nos subgrupos CTL MAOD e DIAB MAOD, os defeitos foram preenchidos com MAOD e nos grupos CTL COAG e DIAB COAG apenas com coágulo sanguíneo. Após 0, 7, 14, 21 e 42 dias, as calotas cranianas foram coletadas para determinação da densidade de volume, número de osteoclastos/mm2 na área do defeito, quantificação por imunoistoquimica e expressão do RNAm para as proteínas RANKL, OPG e TNF-α. Os resultados para volume do tecido ósseo neoformado foi maior nos grupos CTL COAG e CTL MAOD, bem como no grupo DIAB MAOD quando comparado com DIAB COAG (CTL MAOD > CTL COAG e DIAB MAOD > DIAB COAG). O número de osteoclastos nos grupos CTL aumentaram significantemente (3,69 osteoclasto/mm2), enquanto que nos grupos MAOD aumentaram gradualmente até os 42 dias (2,8 osteoclasto/mm2). Os resultados para imunomarcação mostraram que a MAOD promove 1,28 vezes maior expressão de OPG, bem como de TNF-α tanto no grupo CTL (1,59 vezes) como no DIAB (1,76 vezes). Os resultados para expressão do RNAm para OPG mostrou que a média dos valores do grupo COAG comparado com a do grupo MAOD foi 1,91 vezes maior no grupo COAG. Já os valores para expressão de RANKL permaneceram constantes no grupo DIAB MAOD, com aumento significativo de 2,57 vezes aos 42 dias, sendo 4,3 vezes maior, quando comparado com a média dos outros grupos no mesmo período. Conclui-se que nos animais normoglicemicos, o tratamento com a MAOD aumenta a expressão de OPG, RANKL e TNF-α, assim como a atividade osteoclástica, promovendo reabsorção da MAOD e formação de tecido ósseo, enquanto que nos animais diabéticos, a atividade osteoclástica foi reduzida, sem alteração nos níveis de OPG e RANKL, reduzindo a reabsorção da MAOD e consequentemente da formação óssea.(AU)

Participation of osteoclasts was evaluated in reabsorption process of demineralized allogenic bone matrix (DABM) as well as the activity of cytokines RANKL, OPG and TNF- α during formation and bone remodeling in critial size defect of normoglycemic and diabetic rats treated or not with DABM. Therefore, 250 male Wistar rats were used. Thirty rats had femurs and tibias collected and processed to obtain DABM. 220 rats were divided into control group (CTL, n=110) and diabetic group (DIAB, n= 110) injected by a single dose of 47 mg/Kg of body weight streptozotocin. Were made 8mm bone defect on skulls of rats, in subgroups CTL DABM and DIAB DABM, defects were filled with DABM and subgroups CTL CLOT and DIAB CLOT were filled with blood clot. After 0, 7, 14, 21 and 42 days, the skulls were collected to determine the volume density, number of osteoclasts/mm2 into defects area, quantification by immunohistochemistry and RNAm expression of RANKL, OPG and TNF-α cytokines. The results of volume density of newly formed bone was higher in CTL CLOT and CTL DABM, as well as in DIAB DABM compared to DIAB CLOT (CTL DABM > CTL CLOT and DIAB DABM > DIAB CLOT). The number of osteoclasts in CTL groups increased to 3,69 osteoclasts/mm2, while in subgroups treated with DABM gradually increased up until 42 days (2,8 osteoclasts/mm2). Immunohistochemistry showed that DABM promotes an increase of 1.28-fold of OPG expression, as well as TNF-a expression in CTL group (1.59-fold) and DIAB group (1.76-fold). The results of RNAm expression of OPG showed that the average values of the CLOT subgroup compared to the average values of DABM subgroup was 1.91- fold higher in CLOT subgroup. The values of RANKL RNAm expression increase 2.57-fold at 42 days, being 4.3-fold higher than the average os the other groups in the same period. In conclusion, in the normoglicemic animals (CTL group), the treatment with DABM increase the expression of OPG, RANKL and TNF-α as the activity of osteoclasts, leading to DABM resorption and bone tissue formation, while in diabetic animals, the osteoclast activity was reduced, without changes in the leves of OPG and RANKL, decreasing DABM resorption and bone formation.(AU)

Spray-dried extracts from Syzygium cumini seeds: physicochemical and biological evaluation

Aqueous extract from seeds of Syzygium cumini (L.) Skeels, Myrtaceae, obtained by dynamic maceration was spray-dried and characterized by its physico-chemical and antihyperglycaemic action. The extract showed to possess high amount of polyphenols, significant in vitro free radical scavenger activity using the DPPH method and an antihyperglycaemic effect in alloxan-induced experimental diabetes. S. cumini spray-dried extracts were obtained using silicon dioxide and cassava starch as adjuvants. The powders showed acceptable flowability, compactability, and low hygroscopicity at 43% relative humidity. Besides, the spray-dried extracts showed in vivo antihyperglycaemic and in vitro scavenger activity comparable to the lyophilized extract. Thus, experimental data indicates that the extract from S. cumini has a relevant activity and that spray-drying could be adequately used to perform the technological processing of S. cumini fluid extracts.

Effect of Traditional Chinese Medicine Complex for diabetes (TCM-D™) on experimentally induced diabetic mice

Background: We previously evaluated the biochemical changes induced by the local product TCM for diabetes (TCM-D™) on blood glucose levels and other biochemical changes in normal mice fed orally with the recommended human dose (30 ml/kg daily) and ten times this dose for eight weeks. TCM-D™ is an aqueous extract of the roots of Trichosanthes kirilowii Maxim, Paeonia lactiflora Pall, Glycyrrhiza uranlensis Fisch. and Panax ginseng Meyer (red) combined at the dry weight proportions of 36%, 28%, 18% and 18% respectively. The study showed that at these dosages the blood glucose levels as well as the body weights in treated mice were significantly reduced when compared with pretreatment values and control animals. The present study evaluated the effect of the extract in a mouse model of Type 1 diabetes mellitus. Methods: TCM-D™ extract was prepared as a 10x concentrate and given orally at 0.3 ml/100 g and 1.5 ml/100 g to mice which were experimentally induced diabetic with intraperitoneal injections of streptozotocin (5 mg/100g) in sodium citrate (pH 4.5). Control diabetic mice were dosed with extract diluent (distilled water). Results: At the doses studied the compound did not show any significant lowering of the glucose levels in a mouse model of Type 1 diabetes. There were significant increases in the alanine aminotransferase (ALT) and creatinine levels which were most likely due to the treatment with the compound. There were no significant changes in the aspartate aminotransferase (AST) and blood urea levels due to the treatment. Neither was there any significant effect on the weight of the treated animals due to the treatment. Conclusions: It is concluded that TCM-D™ did not have any significant blood glucose lowering effect on streptozotocin induced diabetic mice when fed orally at 1-5 times the recommended human dose. Further work is needed to determine if the extract has any significant effect in a mouse model with Type 2 diabetes mellitus.

Transmission electron microscopy studies of the vestibulocochlear nerve in chronic diabetic rats / Estudios de microscopía electrónica de transmisión del nervio vestibulococlear en ratas diabéticas crónicas

It is widely described in the literature that diabetic patients present hearing impairment. Despite the histological alterations of the internal ear structures in these patients as well as in experimental models of diabetes, to the best of our knowledge, an histological evaluation of the vestibulocochlear nerve have not been performed. In the present study, ultrastructural alterations are described and compared between a spinal nerves and a cranial nerve in rats with chronic induced diabetes. Male Wistar rats (n = 12), fed with standard diet from the animal care facility at 42 days of age were used. Induced diabetic animals (n=6) were fasted for 12 hours prior to being injected intraperitoneally with streptozotocin (STZ - 60mg/kg) in a single dose. Control animals (n=6) received (0.01 mol/l citrate buffer, pH 4.5) vehicle alone. Ten weeks after STZ injection the animals were perfused intracardially with Karnovsky solution. Right and left vestibulocochlear nerves were dissected and histologically processed for epoxy resin embedding. Samples were imaged with the transmission electron microscope. Large myelinated fibers with morphological signs of axonal atrophy in the vestibulocochlear nerves were readily observed. These results suggest that chronic STZ-induced diabetes in rats caused alterations in the myelinated fibers and Schwann cells, compatible to the classic diabetes signs and symptoms. Morphological alterations of the vestibulocochlear nerve in diabetes is described for the first time and contributes information for a better understanding of why there are changes in hearing observed in diabetic patients.

Se ha descrito ampliamente en la literatura que los pacientes diabéticos presentan discapacidad auditiva. En estos pacientes, a pesar de las alteraciones histológicas de las estructuras del oído interno, así como en modelos experimentales de diabetes, que mejoran nuestro conocimiento, la evaluación histológica del nervio vestibulococlear no ha sido realizada. Se describen y comparan las alteraciones ultraestructurales entre un nervio espinal y uno craneal en ratas con diabetes crónica inducida. Fueron utilizadas 12 ratas Wistar machos, de 42 días de edad, alimentadas con dieta estándar. Los animales diabéticos inducidos (n = 6) se mantuvieron en ayuno por 12 horas antes de ser inyectados por vía intraperitoneal con estreptozotocina (STZ - 60mg/kg) en una sola dosis. Los animales control (n = 6) sólo recibieron inyección de 0.01 mol/l buffer, citrato pH 4,5. Diez semanas después de la inyección de STZ, los animales fueron perfundidos intracardiacamente con solución de Karnovsky. Los nervios vestibulococlear derecho e izquierdo fueron disecados y procesados histológicamente para ser incluidos en resina epoxy. Las muestras fueron estudiadas con microscopio electrónico de transmisión. Fueron observadas fácilmente, grandes fibras mielinizadas con signos morfológicos de atrofia axonal en los nervios vestibulococlear. Estos resultados sugieren que la diabetes crónica inducida por STZ en ratas causó alteraciones en las fibras mielínicas y células del neurilema, compatible, con los signos y síntomas clásicos de la diabetes. Alteraciones morfológicas del nervio vestibulococlear en la diabetes son descritas por primera vez, lo que aporta información para una mejor comprensión de por qué hay cambios en la audición en los pacientes diabéticos.

Effect of Tongluo recipe on CD4+ CD25+ regulatory T cells in the thymus of experimental diabetic rats / 第二军医大学学报

Objective: To observe the effect of Tongluo recipe(TLR) on CD4+ CD25+ regulatory T cell in the thymus of experimental diabetic rats. Methods: Totally 112 SD rats were randomly divided into three groups: control group, diabetes mellitus (DM) group, and TLR treatment group. Rat diabetic model was induced by intraperitoneal injection of streptozotocin (60 mg/kg). The TLR treatment group was further divided into two subgroups: prevention group was given intragastric TLR (0.4 g·kg-1·d-1) for 12 weeks starting from the day of streptozotocin injection (TL-1 group), and the treatment group was given intragastric TLR (0.4 g· kg -1·d-1) for 12 weeks starting from 12 weeks after streptozotocin injection (TL-2). The animals were sacrificed at different time points (The control and DM group: 4, 8, 12, 16, 20, and 24 weeks; TLR group: 12 and 24 weeks after administration of TLR). Flow cytometry, H-E staining, and immunochemistry were performed to investigate the change of CD4+ CD25+ regulatory T cell ratio in CD4+ T cells, the thymus morphology, and the expression of Foxp3 protein in the thymus. Results: Compared with the normal control group, diabetic group had a gradually shrunk thymus, a decreased ratio of CD4+ CD25+ regulatory T cells in CD4+ T cells, and a decreased expression of Foxp3. The ratios of CD4+ CD25+ regulatory T cells and the thymus indices in the two TLR groups were significantly higher than those in the diabetic model group(P<0.05). Conclusion: Tongluo recipe can greatly increase the ratio of CD4+ CD25+ regulatory T cells, thymus index, and Foxp3 expression in the thymus, and may protect the thymus CD4+ CD25 + regulatory T cells in experimental diabetic rats.

Avaliação do reparo de defeito ósseo em calvária de ratos diabéticos tratados com matriz óssea desmineralizada / Evaluation of repair of bone defects in skulls of diabetic rats treated with demineralized bone matrix

O objetivo deste trabalho foi avaliar as atividades osteoindutoras e osteocondutoras da matriz alogênica óssea desmineralizada (MAOD) frente à diabetes no reparo de defeito de tamanho crítico em calvárias de ratos diabéticos. Para isso, 100 ratos machos Wistar foram divididos em 2 grupos: no grupo diabético (DIAB, n=50) foi injetado 47 mg/Kg de massa corporal de estreptozotocina, enquanto que no grupo controle (CTL, n=50) foi injetado solução fisiológica a 0,9%. A MAOD foi obtida de 50 ratos, cujo fêmur e tíbia foram retirados, desmineralizados com HCl a 0,6M por 24 horas, particulados em 1-2mm³, neutralizados com soro fisiológico e armazenados em álcool. Após a anestesia, foram realizados defeitos ósseos de 8 mm nas calvárias dos animais, sendo os grupos CTL COAG (n=25) e DIAB COAG (n=25) preenchidos com coágulo e os grupos CTL MAOD (n=25) e DIAB MAOD (n=25) preenchidos com MAOD. Após os períodos de 0, 7, 14, 21 e 42 dias, as calvárias foram coletadas. A análise radiográfica mostrou que houve formação de ilhas radiodensas no interior dos defeitos nos grupos CTL e DIAB tratados com MAOD, enquanto que nos grupos tratados com coágulo houve formação de áreas mais radiodensas somente nas bordas do defeito, corroborando com os resultados morfológicos, que mostraram nos grupos tratados com coágulo que o reparo ósseo teve início nas bordas do defeito, enquanto que nos grupos tratados com MAOD, a neoformação óssea ocorreu também nas áreas de reabsorção nas partículas de MAOD. De acordo com os resultados morfométricos, o volume de tecido ósseo aumentou gradativamente em todos os grupos, porém, esse aumento foi maior nos grupos CTL em relação aos seus respectivos tratamentos nos grupos DIAB (CTL COAG > DIAB COAG e CTL MAOD > DIAB MAOD) e maior quando comparados os grupos tratados com MAOD versus os respectivos grupos tratados com COAG (CTL MAOD > CTL COAG e DIAB MAOD > DIAB COAG). Assim, ao término de 42 dias, o volume de tecido ósseo no grupo CTL MAOD foi...

The aim of this work was to evaluate the osteoinductive and osteoconductive activities of demineralized allogeneic bone matrix (DABM) against diabetes in repairing critical size defects in diabetic rats skulls. Therefore, 100 male Wistar rats were shered into two groups: in the diabetic group (DIAB, n=50) was 47 mg/Kg of body weight streptozotocin, while in the control group (CTL, n=50) was injected saline 0.9%. The DABM was obteined using 50 rats which were removed their femur and tibia bones, demineralized in 0.6 N HCl during 24 hours, cut into 1-2mm³ pieces, neutralized in saline and stored in alcohol. After anesthesia, were made 8 mm bone defects on skulls of rats, being the CTL CLOT group (n=25) and DIAB CLOT group (n=25) filled with blood clot and the CTL DABM group (n=25) and DIAB DABM group (n=25) filled with DABM. After 0, 7, 14, 21 and 42 days, the skulls were collected. The radiographic analysis showed radiodense islets inside the defects filled with DABM in CTL and DIAB groups, while groups filled with blood clot showed radiodense areas near the defect border, which is in agreement to the morphologic results, that had showed the begining of bone healing was near the defects border in groups filled with blood clot, while groups filled with DABM showed new bone formation also in resorption DABM areas. According to morphometric results, the volume of bone tissue had increased in all groups, however, this increase was more accentuated in CTL groups when compared to DIAB groups with respected treatments (CTL CLOT > DIAB CLOT and CTL DABM > DIAB DABM) and bigger when groups treated with DABM are compared to respestive groups treated with CLOT (CTL DABM > CTL CLOT e DIAB DABM > DIAB CLOT). Thereby, at the end of 42 days, the CTL DABM bone tissue volume was 3.24 greater than the other groups, the CTL CLOT and DIAB DABM groups didnt show any significant differenceand the DIAB DABM was 1,81 greater than DIAB CLOT. From these results, the conclusion...

The diabetes-induced functional and distributional changes of the alpha 1-adrenoceptor of the abdominal aorta and distal mesenteric artery from streptozotocin-induced diabetic rats / 대한마취과학회지

BACKGROUND: The aim of this study was to evaluate the effect of diabetes on the function and distribution of vascular alpha1-adrenoceptors in the abdominal aorta and distal mesenteric artery from streptozotocin (STZ)-induced diabetic rats at the level of the alpha1-adrenoceptor subtypes. METHODS: Diabetes was induced by a single intravenous injection of STZ (60 mg/kg) in 8 week-old male Sprague-Dawley rats (n = 11). Age-matched normal rats (n = 14) were used as a control group. Four weeks after STZ injection, the tilting-induced change of the mean arterial pressure was recorded. The alpha1-adrenoceptor subtypes mediating the contractions of the distal mesenteric artery and abdominal aorta were investigated using the agonist phenylephrine and subtype-selective antagonists that included prazocin, 5-methylurapidil and BMY 7378. The expressions of the alpha1-adrenoceptor subtypes of each artery were examined by immunofluorescence staining using the subtype selective antibodies. RESULTS: The recovery of the mean arterial pressure was delayed after positional change in the diabetic rats. Compared with that of the normal rats, the contractile response to phenylephrine was increased in the abdominal aortas and it was decreased in the distal mesenteric arteries in the diabetic rats. In addition, compared with the normal rats, the fluorescent intensity of all the alpha1-adrenoceptor subtypes was increased in the abdominal aortas and it was decreased in the mesenteric arteries of the diabetic rats. CONCLUSIONS: Diabetes increased the contractility of the abdominal aorta in response to phenylephrine, yet diabetes decreased that of the mesenteric arteries in the STZ-induced diabetic rats. Those results are mainly based on the overall change of the alpha1-adrenoceptor, and not on the change of the specific alpha1-adrenoceptor subtypes.

A microcirculação da bolsa da bochecha do hamster sob a influência do diabetes mellitus experimental induzido por estreptozotocina: aspectos morfofuncionais / Microcirculation of hamster cheek pouch under the influence of experimental Diabetes Mellitus induced by streptozotocin: morpho-functional aspects

O Diabetes Mellitus é uma doença metabólica crônica com múltiplos fatores etiológicos (genético, viral e imunológico) que condiciona deficiência absoluta ou relativa de insulina, causando persistência de níveis elevados de glicose no sangue. Atualmente, o Diabetes Mellitus é considerado um importante problema de saúde devido a sua prevalência e alta morbimortalidade. Sua importância clínica resulta essencialmente de suas graves complicações, especialmente as microvasculares. A hiperglicemia crônica ou intermitente tem sido identificada como o fator indutor de lesão endotelial, sendo este, o agente desencadeante das complicações microvasculares. As células endoteliais, por serem influenciadas pela força hemodinâmica local, respondem com a transdução de sinais (mecanotrans dução), as quais podem ser responsáveis pelo início de processos patológicos na parede dos vasos. Desta forma, o objetivo deste estudo foi analisar a microcirculação da bolsa da bochecha do hamster sob a influência do Diabetes Mellitus tipo 1 experimental induzido por estreptozotocina, avaliando seus aspectos morfofuncionais aos 6 e 15 dias de evolução da doença. As características morfológicas de arteríolas e vênulas foram estimadas por medidas do diâmetro do lúmen e da espessura da parede; pela densidade de volume e de área destes vasos na bolsa da bochecha; pela análise imunohistoquímica da expressão de actina, talina, alfa-actina de músculo liso, vimentina, laminina e colágeno IV através da microscopia de luz com a utilização de um sistema semiquantitativo baseado em uma escala de intensidade de imunomarcação; e por microscopia eletrônica de transmissão. Também foi avaliado o relaxamento dependente do endotélio, medido pela variação do diâmetro do lúmen antes e após a aplicação de acetilcolina e a permeabilidade de vênulas pós-capilares à histamina, determinada pelo número de pontos de extravasamento plasmático. Nossos resultados mostraram que arteríolas e vênulas...

Diabetes Mellitus is a chronic metabolic disease with multiple etiologic factors (genetic, viral and immunological) that results in absolute or relative insulin deficiency, causing persistent elevated blood glucose levels. Nowadays, Diabetes Mellitus is considered as an important health concern due to its increasing prevalence and high morbimortality. Its clinical importance comes from the complications, especially harm inductor factor, being this the first outcome of microvascular complications. Endothelial cells, under local hemodynamic strength, produce signal transduction (mechanotransduction), which can be responsible for the beginning of patholgical events in vessels wall. In this regard, the objective of this study was to analyze hamster cheek pouch microcirculation under the influence of type 1 diabetes mellitus induced by streptozotocin, evaluationg its morpho-functional aspects at 6 and 15 days of diseases evolution. Morphological characteristics of arterioles and venules were estimated by the measurement of lumen diameter and wall thickness; the volume density and area of these vessels from cheek pouch; immunohistochemistry of the expression of actin, talin, smooth muscle alpha-actin, vimentin, laminin and type IV collagen through light microscopy with the utilization of a semi-quantitative score system based on the intensity of the immunostaining; and transmission electron microscopy. It was also evaluated the endothelium dependent relaxation, measured by the variation of lumen diameter before and after acetylcholine administration and post-capillary venules permeability to histamine, determined by number of points of plasma extravasation. Our results reveal that arterioles and venules do not show differences between the groups concerning wall thickness, luminal diameter, density per area and volume density. Vascular permeability, after 2 minutes of histamine administration, was reduced significantly in diabetic groups...

Repeated oral treatment with polysaccharide sulfate reduces insulin resistance and dyslipidemia in diabetic dyslipidemic rat model / 药学学报

Polysaccharide sulfate (PSS) is a new type of antiatherosclerotic medicine for its effects of anticoagulation, anti-thrombosis and modulation of dyslipidemia. However, it is still uncertain whether PSS could modulate the diabetic dyslipidemia or not. Here, the rat model of diabetic dyslipidemia was developed and the effects of PSS on glucose and lipid levels were investigated in this animal model. Wistar rats were iv injected with streptozotocin 20 mg·kg-1 after feeding with high fat diet for one and a half month. Then, rats received orally PSS (30, 90, and 180 mg·kg-1) for 1 month. After oral treatment with PSS (90 and 180 mg·kg-1) for 1 month, the levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C) were significantly reduced and the level of high density lipoprotein-cholesterol (HDL-C) increased, compared with diabetic control rats. Moreover, PSS (30, 90, and 180 mg·kg-1) had a tendency to reduce glucose and insulin levels, and significantly increased insulin sensitivity index. Our results suggest that PSS could improve insulin sensitivity and relieve dyslipidemia in diabetic dyslipidemic rats.

Effect of Antioxidants on Myocardial Damage in Streptozotocin-Induced Diabetic Rats

BACKGROUND AND OBJECTIVES: Many diabetic patients suffer from cardiomyopathy, even in the absence of vascular disease. The aim of this study was to see if dietary antioxidant supplementation has an inhibitory effect on the progression of cardiac tissue damage in streptozotocin (STZ)-induced diabetic rats. MATERIALS AND METHODS: Sprague-Dawley male rats (n=60) were used as experimental animals; they were divided into the normal control group and the diabetic group. Eight weeks after STZ injection (65 mg/kg of body weight), the products of lipid peroxidation (malondialdehydes, MDA), and the antioxidant enzyme superoxide dismutase (SOD), and catalase activities were determined in the cardiac tissue homogenates. The cardiac tissues were studied by light microscopy (LM) and electron microscopy (EM), and the tissue lesions were graded by a semiquantitative score. RESULTS: The histologic scores for perivascular fibrosis, interstitial fibrosis and myocardial necrosis according to LM were significantly lower in the combined vitamin C & E treated rats than in the diabetic control rats. The ultrastructural scores for the overall cardiac morphology, mitochondria and myofilaments, according to EM, were significantly lower in the vitamin E treated rats and the combined vitamin C & E treated rats than in the diabetic control rats, even though this was of less magnitude than that in the insulin-treated diabetic rats. CONCLUSION: These results suggest that antioxidants such as vitamin C & E might have a beneficial effect on diabetes as an adjunct therapy against lipid peroxidation and diabetic cardiomyopathy, in addition to the effects of instituting strict measures for controlling the blood glucose.

Suppression of retinal blood vessel leakage by inhibition of vascular endothlial growth factor using soluble-Flt in experimental diabetic rats / 眼科新进展

Objective　Vascular endothelial growth factor (VEGF) can cause potent vascular permeation. More and more findings have demonstrated that VEGF is increased in the diabetic retinopathy. The objective of this study is to investigate whether blocking the VEGF using soluble-Flt can suppress the retinal blood vessel leakage.Methods　Sprague-Dawley rats weighing approximately 200 grams were made diabetic by injection of streptozotocin (STZ). After the animals were diabetic for 1 week, they were intravenously injected with either PBS+50g*L-1Glycerol, interleukin-6 receptor monoclonal antibody (IL-6R), or soluble-Flt. On the eighth day, Evans blue method was used to check the leakage. Animals treated with citrate sodium buffer and STZ treated non-diabetic animals served as the control group. Results　There was no significant difference of the retinal blood vessel leakage between the two control groups (P>0.05). The retinal blood vessel leakage of 8 day diabetic animals was significantly higher than that of total control groups (P<0.05). The leakage of diabetic animals treated with PBS+Glycerol or IL-6R remains the same (P>0.05). But, the leakage of 25 mg*kg-1 soluble-Flt treated diabetic animals was remarkably lower than that before treatment, PBS+Glycerol treated or IL-6R treated diabetic animals (P<0.000 1), and there was no difference compared with total non-diabetic control groups (P>0.05). Conclusion　This study shows that the VEGF plays a very important role in the onset of the diabetic retinopathy and suggests by suppressing the action of VEGF, the retinal blood vessel leakage at the early stage of diabetic retinopathy could be reduced.

Carbohydrates, lipids and lipoproteins and islet changes in diabetes with superimposed myocardial infarction.

Short-term metabolic and concomitant morphologic effects of streptozotocin diabetes on isoproterenol-induced myocardial infarction was studied in Wistar rats, Of particular significance was the observation that myocardial infarction in concert with diabetes brought about a distinctive exacerbation of the severity and complexity of the histopathological lesions. Of all the biochemical parameters, serum glucose and free fatty acids registered maximum elevation and serum lactate and cardiac glycogen levels a maximum reduction. Among the lipoproteins, an inverse relationship was found between high density lipoproteins and low density and very low density lipoproteins; while high density lipoproteins, ratio of high density lipoprotein to low density lipoprotein and the percentage of high density lipoprotein were decreased, there was a significant increase in low density lipoprotein concentration and percentage values of low density and very low density lipoproteins. In diabetes, the Β cell of the endocrine pancreas depicted selective necrosis. Loss of insulin granules and wide-spread necrobiosis of cellular elements of the pancreatic islets were observed, respectively, in myocardial infarction and in diabetes plus myocardial infarction combinations. Pathological evidence of chemical-induced mild toxicity was present in the exocrine parenchyma. Mitotic features and the presence of centroacinar cells in the damaged Langerhans’ islets supposedly formed the basis of regeneration of the tissue in diabetes, with or without vascular complications.

THE INFLUENCE OF EXPERIMENTAL DIABETES ON THE EPITHELIAL CELLS IN RAT THYMIC CORTEX / 解剖学报

Objective To explore the morphologic changes of the epithelial cells in the thymic cortex in diabetes rats, and its possible mechanism of thymocyte apoptosis and thymus involution. Methods The ultrastructure changes of the epithelial cells in the thymic cortex in diabetes rats were observed under electron microscope. Results The type 2 epithelial cells had four kinds of the changes on ultrastructure:1 the endocrine disorder like changes with a large number of the degenerative cystic vacuoles;2 the degenerative like changes with lots of the abnormal secretory vacuoles and myelin like bodies and bulky tonofilament bundles; 3 the phagocyte like changes with a great number of the dense lysosome like granulae and the phagocytosed apoptotic cells and bodies; 4 the apoptosis like changes with the early morphologic changes of apoptosis. The type 3 epithelial cells had two kinds of the changes on ultrastructure:1 degenerative like changes with groups of dense granulae which secrete peptide hormones;2 the proliferative like changes with numerous collage fibrils in the vicivity of their perikarya. In addition, a lot of the apoptotic thymocytes and the reduction of thymocyte number were found in thymic cortex.Conclusion The varied ultrastructure changes of the epithelial cells in rat thymic cortex could be induced by diabetes, which might be the one of major mechanisms on influencing the thymocyte development and causing the thymocyte apoptosis and thymus involution.\;[