Effect of docosahexaenoic acid intake on lipid peroxidation in diabetic rat retina under oxidative stress

Docosahexaenoic acid (DHA) plays an important role in visual function but has a highly oxidation-prone chemical structure. Therefore, we investigated how dietary DHA affects the generation of lipid peroxides in rat retina under oxidative stress in diabetes with/without vitamin E (VE) deficiency. Streptozotocin-induced (50 mg i.p./kg B.W.) diabetic Sprague-Dawley (SD) rats were assigned to four groups: (i) control/VE(+), (ii) DHA/VE(+), (iii) control/VE( - ) and (iv) DHA/VE( - ), and raised for 28 days. We then measured lipid peroxide levels in the retina, serum and liver. With a normal intake of VE, dietary DHA increased only the retinal level of thiobarbituric acid-reactive substances (TBARS) slightly. In contrast, in rats with VE deficiency, dietary DHA increased serum and liver lipid peroxide levels but not in the retina. These results suggest that dietary DHA does not necessarily promote lipid peroxidation in the retina even under high oxidative stress.     

Alterations in the Retinal Dopaminergic Neuronal System in Rats with Streptozotocin-Induced Diabetes

Neurochemical alterations, which may be associated with the development of diabetic retinal dysfunction, were investigated using streptozotocin (STZ)-induced hyperglycemia in rats. Young male Wistar rats, weighing 100-150 g, were made diabetic with daily intraperitoneal injections of STZ (30 mg/kg) for 5 days. This treatment caused a continuous hyperglycemia (400-600 mg/dl) and suppressed gain in body weight. Nine weeks after the STZ treatment, a significant increment in retinal valine and a decline in phenylalanine were noted, while the concentrations of other neuroactive amino acids, such as gamma-aminobutyric acid and aspartic acid, in the retina remained unchanged. On the other hand, the concentration of retinal dopamine (DA) was found to decrease significantly from the third week of hyperglycemia, when [3H]spiperone binding showed a tendency to increase in the retinal particulate fraction. However, the activities of tyrosine hydroxylase and aromatic L-amino acid decarboxylase (AADC) and the uptake of [3H]tyrosine showed no alteration in the retina of diabetic rats. The accumulation rate of 3,4-dihydroxyphenylalanine (DOPA) in vivo in the retina of diabetic rats, measured following the administration of the AADC inhibitor m-hydroxybenzyl-hydrazine (100 mg/kg i.p.), was also unchanged. Although [3H]DA uptake by retinal tissue was similar in control and diabetic animals, the spontaneous efflux of [3H]DA from the retina was found to be significantly accelerated in STZ-treated animals. In addition, the release of preloaded [3H]DA, elicited by repeated photic stimulation, was significantly attenuated in retina from diabetic rats. These results suggest that an accelerated efflux of DA, possibly leading to the depletion of DA from the retinal DA system, may account for early retinal dysfunctions known to occur in diabetic subjects.     

Retinal glutamate in diabetes and effect of antioxidants

Diabetes results in various biochemical abnormalities in the retina, but which of these abnormalities are critical in the development of retinopathy is not known. The aim of this study is to examine the effect of antioxidant supplementation on diabetes-induced alterations of retinal glutamate, and to explore the inter-relationship between alterations of retinal glutamate, oxidative stress, and nitric oxide (NO) in diabetes. Glutamate was measured in the retina at 2 months of diabetes in rats receiving diets supplemented with or without a mixture of antioxidants containing ascorbic acid, Trolox, DL alpha-tocopherol acetate, N-acetyl cysteine, beta-carotene and selenium. The relationship between glutamate, oxidative stress and NO was evaluated using both bovine retinal endothelial cells and normal rat retina. In diabetes, retinal glutamate was elevated by 40, thiobarbituric acid-reactive substances (TBARS) by 100, and NO by 70%, respectively. Administration of antioxidants inhibited the diabetes-induced increases in glutamate, TBARS and NO. Incubation of bovine retinal endothelial cells or normal rat retina with glutamate significantly increased TBARS and NO, and addition of either antioxidant (N-acetyl cysteine) or a NO synthase inhibitor prevented the glutamate-induced elevation in oxidative stress and NO. Incubation of retina with a glutamate agonist, likewise elevated oxidative stress and NO, and memantine inhibited such elevations. Thus, the alterations of retinal glutamate, oxidative stress and NO appear to be inter-related in diabetes, and antioxidant therapy may be a suitable approach to determine the roles of these abnormalities in the development of diabetic retinopathy.     

Antioxidant role of Umbelliferone in STZ-diabetic rats

The objective of the study was to investigate the role of Umbelliferone (UMB) on lipid peroxidation, nonenzymic and enzymic antioxidants in the plasma and liver of streptozotocin (STZ)-induced diabetic rats. Adult male albino rats of Wistar strain, weighing 180-200 g, were induced diabetes by administration of STZ (40 mg/kg b.wt.) intraperitoneally. The normal and diabetic rats were treated with UMB (30 mg/kg b.wt.) dissolved in 10% dimethyl sulfoxide (DMSO) for 45 days. Diabetic rats had an elevation in the levels of lipid peroxidation markers (thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides (HP) and conjugated dienes (CD)), and a reduction in nonenzymic antioxidants (vitamin C and reduced glutathione (GSH) except vitamin E in the plasma and liver, and enzymic antioxidants (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in the liver. Decreased level of beta-carotene and increased level of ceruloplasmin (Cp) were observed in the plasma of diabetic rats. Treatment with UMB and glibenclamide brought back lipid peroxidation markers, nonenzymic and enzymic antioxidants to near normalcy. Since UMB treatment decreases lipid peroxidation markers and enhances antioxidants' status it can be considered as a potent antioxidant.     

Changes in Lipid Peroxide Level in Retinal Pigment Epithelial Cells In Vitro Upon Addition of Linoleic Acids or Linoleic Acid Hydroperoxides Under Varying Concentrations of Oxygen

We previously observed the presence of autofluorescent lipofuscin or its like in retinal pigment epithelial (RPE) cells, which were incubated with linoleic acid hydroperoxides (LHP). We studied the effect of oxygen on the level of lipid peroxides in RPE cells in the presence of linoleic acids (LA) or LHP. The level of lipid peroxides in these cells was determined by use of the thiobarbituric acid-reactive substance (TBARS), which responded to oxygen concentrations qualitatively, and a linear regression analysis. Multiple linear regression analysis disclosed that treatment with LA for 24 hr resulted in detectable increase in the level of TBARS in the cells, whereas treatment with LA or LHP for 48 hr caused detectable decrease. Stepwise linear regression analysis showed that the level of TBARS decreased in an oxygen-tension dependent manner in the cells incubated with LA for 48 hr. Thus, it was shown that short-term incubation with LA increased the level of TBARS in the cells and that LA decreased its level in an oxygen-tension dependent manner. For these results, the postulation was made that the prolonged auto-oxidation of LA caused production of lipofuscin-like materials, a complex of lipid peroxides and proteins that were insoluble in SDS and acetic acid solution.     

Effects of leptin on oxidative stress in healthy and Streptozotocin-induced diabetic rats

Aims/hypothesis It is generally accepted that oxidative stress is responsible for etiology and complications of diabetes. During uncontrolled Type 1 diabetes, plasma leptin levels rapidly fall. However, it is not known whether diabetes-induced hypoleptinemia has any role in oxidative stress related to uncontrolled Type I diabetes. The present study was designed to examine the effects of leptin treatment on plasma lipid peroxidation and reduced glutathion of normal and streptozotocin(STZ)-induced diabetic rats. Methods Diabetes was induced by single injection of Streptozotocin (55 mg/kg bw). One week after induction of diabetes, rats began 5-day treatment protocol of leptin injections of (0.1 mg/kg bw i.p.) or same volume vehicle. At the end of the 5th day, rats were sacrificed by cardiac puncture under anesthesia and their plasma was taken for plasma leptin, malondialdehyde, and reduced glutathione measurements. Results Plasma leptin levels decreased in STZ-induced diabetic rats while plasma glucose, TBARS, and GSH levels increased. Plasma leptin levels were not affected with leptin treatment in both diabetic and non-diabetic rats. The elevation in plasma TBARS associated with STZ diabetes decreased with leptin treatment. Leptin also increased plasma GSH levels in diabetic rats. In non-diabetic rats, treatment with leptin did not change plasma TBARS and GSH levels. Conclusions/interpretations In conclusion, leptin treatment is able to attenuate lipid peroxidation in STZ-diabetic rats, in the onset of diabetes, by increasing the GSH levels without affecting hyperglycemia and hypoleptinemia.     

Ameliorative potential of S-allyl cysteine on oxidative stress in STZ induced diabetic rats

Increased oxidative stress and impaired antioxidant defense mechanism are important factors in the pathogenesis and progression of diabetes mellitus and other oxidant-related diseases. The present study was undertaken to evaluate the possible protective effects of S-allyl cysteine (SAC) against oxidative stress in streptozotocin (STZ) induced diabetic rats. SAC was administered orally for 45 days to control and STZ induced diabetic rats. The effects of SAC on glucose, plasma insulin, thiobarbituric acid reactive substances (TBARS), hydroperoxide, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH), oxidized glutathione (GSSG) and GSH/GSSG ratio were studied. The levels of glucose, TBARS, hydroperoxide, and GSSG were increased significantly whereas the levels of plasma insulin, reduced glutathione, GSH/GSSG ratio, superoxide dismutase, catalase and GPx were decreased in STZ induced diabetic rats. Administration of SAC to diabetic rats showed a decrease in plasma glucose, TBARS, hydroperoxide and GSSG. In addition, the levels of plasma insulin, superoxide dismutase, catalase, GPx and reduced glutathione (GSH) were increased in SAC treated diabetic rats. The above findings were supported by histological observations of the liver and kidney. The antioxidant effect of SAC was compared with glyclazide, a well-known antioxidant and antihyperglycemic drug. The present study indicates that the SAC possesses a significant favorable effect on antioxidant defense system in addition to its antidiabetic effect.     

早期糖尿病大鼠晶状体和视网膜水通道蛋白4表达的研究

目的观察早期糖尿病大鼠晶状体、视网膜水通道蛋白4(aquaporin 4,AQP-4)表达的变化,探讨糖尿病大鼠眼组织水代谢改变的机制。方法 SD大鼠分为正常对照组和糖尿病组。制作糖尿病大鼠模型,于第4、8周取材,用免疫组化法和计算机图像分析系统半定量分析各组大鼠晶状体、视网膜AQP-4表达的变化。结果正常及糖尿病大鼠AQP-4在晶状体上皮均无表达。AQP-4在视网膜上有表达,正常大鼠主要表达于视网膜视杆视锥层、节细胞层和神经纤维层,糖尿病大鼠从内界膜延伸至视细胞层整个视网膜厚度均可见AQP-4阳性表达,特别是在神经节细胞层毛细血管内皮和神经纤维层阳性表达更明显。糖尿病大鼠视网膜组织AQP-4阳性表达标随周龄的延长而增强。结论糖尿病大鼠视网膜AQP-4的表达较正常组增强,提示水通道蛋白的表达增加是糖尿病早期发生视网膜水肿的机制之一。     

血小板反应素1在STZ诱导大鼠糖尿病性视网膜病变的表达研究

探讨TSP1表达在糖尿病视网膜病变中的作用和机制,为治疗和预防糖尿病视网膜病变提供新的实验和理论依据。用链脲佐菌素(STZ)腹腔注射建立糖尿病模型8周后,采用免疫组织化学、RT-PCR及实时荧光定量PCR法,分析TSP1在早期链尿佐菌素诱导的糖尿病SD大鼠视网膜中的表达。结果显示在早期糖尿病大鼠的视网膜表面血管、神经节细胞层、内外核层中均有明显的TSP1表达,糖尿病视网膜组TSP1 mRNA表达要高于对照组,其中实时荧光定量PCR CT值的结果显示糖尿病组TSP1 mRNA表达量较对照组要高约3．48倍,二组间差别有显著性意义(P<0．01),提示TSP1在视网膜组织中的表达与糖尿病视网膜病变的发生密不可分,TSP1表达的增加可能在糖尿病视网膜病变的发生和发展中起重要作用。     

Des(1–3)IGF-1 Treatment Normalizes Type 1 IGF Receptor and Phospho-Akt (Thr 308) Immunoreactivity in Predegenerative Retina of Diabetic Rats

Little is known about interventions that may prevent predegenerative changes in the diabetic retina. This study tested the hypothesis that immediate, systemic treatment with an insulin-like growth factor (IGF)-1 analog can prevent abnormal accumulations of type 1 IGF receptor, and phospho-Akt (Thr 308) immunoreactivity in predegenerative retinas of streptozotocin (STZ) diabetic rats. Type 1 IGF receptor immunoreactivity increased approximately 3-fold in both inner nuclear layer (INL) and ganglion cell layer (GCL) in retinas from STZ rats versus nondiabetic controls. Phospho-Akt (Thr 308) immunoreactivity increased 5-fold in GCL and 8-fold in INL of STZ rat retinas. In all cases, immunoreactive cells were significantly reduced in STZ des(1–3)IGF-1–treated versus STZ rats. Preliminary results suggested that vascular endothelial growth factor (VEGF) levels may also be reduced. Hyperglycemia/ failure of weight gain in diabetic rats continued despite systemic des(1–3)IGF-1. These data show that an IGF-1 analog can prevent early retinal biochemical abnormalities implicated in the progression of diabetic retinopathy, despite ongoing hyperglycemia.     

Impact of umbelliferone on erythrocyte redox status in STZ-diabetic rats

Oxidative stress is currently hypothesized to be a mechanism underlying diabetes. The present study was designed to evaluate the effect of umbelliferone (UMB), a derivative of coumarin, on erythrocyte lipid peroxidation, antioxidants, and lipid profile in normal and streptozotocin (STZ) diabetic rats. Diabetes was induced in adult male albino rats of Wistar strain, weighing 180 to 200 g, by the administration of STZ (40 mg/kg/b-wt) intraperitonially. The normal and diabetic rats were treated with UMB in 10 percent dimethyl sulfoxide (DMSO) dissolved in water for 45 days. The diabetic rats had elevated levels of blood glucose and lipid peroxidation markers such as thiobarbituric acid reactive substances (TBARS), conjugated dienes (CD), and lipid hydroperoxide (HP) and decreased levels of nonenzymatic antioxidants (Vitamin C and reduced glutathione [GSH]), elevated levels of vitamin E, and elevated levels of enzymatic antioxidants (superoxide dismutase [SOD], catalase [CAT], glutathione peroxidase [GPx]), elevated glucose-6-phosphate dehydrogenase activity, and altered lipid profile (cholesterol and phospholipids) in erythrocytes. These changes were reversed by treatment with UMB. Thus, our results indicate that the administration of UMB shows promising potential for the restoration of normal blood glucose levels, erythrocyte lipid peroxidation, antioxidants, and lipid profile in STZ-diabetic.     

Effects of vitamin A and insulin on the antioxidative state of diabetic rat heart: a comparison study with combination treatment

Because elevated oxidative stress may exacerbate cardiovascular complications of diabetes mellitus, the current study aimed to investigate the effects of treatment with either vitamin A, an antioxidant, or with insulin on lipid peroxidation products and antioxidant enzyme activities of diabetic rat heart. Also to evaluate whether a combination of vitamin A and insulin exerts more beneficial effects than treatment with each agent alone. Rats were made diabetic with a single injection of streptozotocin (STZ, 55 mg kg(-1) i.p.). Two days after STZ-injection, one group of diabetic rats was treated with vitamin A (retinol acetate, 30 mg kg(-1) day(-1) i.o.) for 12 weeks. A second group of diabetic rats was untreated for 6 weeks and then treated for another 6 weeks with insulin (8-10 IU rat(-1) day(-1) s.c.). Both therapies were applied to another group of diabetic rats for assessment of combined therapy with vitamin A plus insulin. Hearts from 12-week untreated diabetic animals showed about a four-fold increase in the level of thiobarbituric acid reactive substances (TBARS), indicative of increased lipid peroxidation. This was accompanied by approximately 100% increase in both catalase and glutathione peroxidase (GSHPx) enzyme activities. Therapy with insulin alone caused a small but significant improvement in plasma TBARS as well as GSHPx activities, but no significant change in plasma catalase in diabetic animals. Diabetes-induced disturbance in TBARS was almost completely prevented by vitamin A therapy. Although, a similar degree of activities for GSHPx was determined in diabetic animals treated with each agent alone, combination therapy was found to be more effective than single therapies in the recovery of GSHPx of diabetic heart. In contrast to insulin single therapy, vitamin A alone significantly prevented an increase in catalase activity of diabetic heart, and a combination of these agents did not supply any further benefit. Superoxide dismutase (SOD) activity was not found significantly different among the experimental groups. STZ-diabetes also resulted in less plasma retinol and retinol-binding protein (RBP), which was significantly improved by insulin single therapy while vitamin A used alone, failed to increase plasma retinol and RBP levels of diabetic animals. Our findings suggest that single therapy with insulin is unable to preclude oxidative reactions in diabetic heart to the same extent as obtained by vitamin A therapy alone, in spite of allowing recovery of normal growth rate and improved vitamin A metabolism in diabetic rats. A combination of insulin with vitamin A may provide more benefits than use of either agent alone in the treatment of general characteristics of diabetes and the maintenance of antioxidant defence of diabetic heart and thus in the reduction of peroxidative stress-induced cardiac injury.     

Outer retinal anoxia during dark adaptation is not a general property of mammalian retinas

The purpose of this study was to measure the intraretinal oxygen distribution across the retina under conditions, which maximise outer retinal oxygen consumption. In particular, we looked for evidence of increased oxygen delivery from the choroid and the deep retinal capillary layer, and whether or not this was sufficient to avoid the development of intraretinal anoxia. Under dark-adapted conditions the photoreceptors need additional energy, at least part of which is derived from increased oxidative metabolism. In earlier studies in the cat retina it was revealed that dark adaptation could render some regions of the outer retina anoxic. The present study of the in vivo oxygen distribution across the rat retina in light and dark found no evidence of outer retinal anoxia in the dark. This was despite a mean increase of 52.6+/-11.4% (n=7) in outer retinal oxygen consumption in the dark. The mean value for the minimum outer retinal PO(2) in the dark was 5.2+1.2 mmHg. Oxygen delivery from both the choroid and the deep retinal capillary layer increased in the dark (P<0.01, and P<0.001, respectively). It is argued that the ability of the deep capillary layer to compensate for changes in oxygen demand in the outer retina is an important element in the maintenance of homeostasis in the retina. This is in addition to the role of the deep capillary layer in supplying oxygen to the highly consuming plexiform layers within the inner retina. These findings in the rat retina also demonstrate that intraretinal anoxia in the dark, is not, as implied by earlier work in the cat, a general feature of mammalian retinas.     

The effects of aminoguanidine on retinopathy in STZ-induced diabetic rats

BackgroundThe accumulation of advanced glycated end products (AGEs) in retinal blood vessels is one of the major etiological factors contributing to diabetic retinopathy. Aminoguanidine (AG) is one of the most extensively used inhibitors of AGEs formation. The aim of this study was to investigate whether AG could protect the development of diabetic retinopathy through inhibition of AGEs.MethodsRat diabetes was induced by intraperitoneal injection with streptozotocin (STZ). AG was given to rats in drinking water. Retina was extracted 3 and 6 months following STZ and AG administration. Immunochemistry and transmission electron microscope were used to detect the expression of AGEs and retina morphology.ResultsExtensive staining of AGEs was detected in retinal blood vessels of 3- and 6-month diabetic rats, while no significant staining was found in the control non-diabetic retina or AG treated groups. Pericyte loss, endothelial cell proliferation, increased ratio of endothelial cells/pericytes, acellular capillaries and capillary occlusion were observed in the retina of 6-month diabetic rats. The increased electron density of retinal capillary basement membrane, mitochondrial swelling in pericytes and endothelial cells were also found in 6-month diabetic rats. The 3-month diabetic rats and the AG-treated rats did not have similar morphological changes compared to control group. The AGEs staining in AG-treated rats was still weakly positive.ConclusionsAGEs plays pivotal roles in diabetic retinopathy. AGE deposition occurs prior to retinal microvasculature changes. AG could prevent the onset and development of diabetic retinopathy through inhibition of AGEs.     

Lipid peroxidation products in human subretinal fluid

The concentrations of thiobarbituric acid reacting substances (TBARS) and proteins in the subretinal fluid (SF) of patients undergoing retinal detachment surgery have been determined. We have tried to establish the correlations between these biochemical and other clinical features of these patients: evolution time of the retinal detachment, age, degree of myopia, and macular affection. Caucasian patients, 19 men and 19 women (57.42 ±12.85 average age, interval 20–80) were randomly selected for this study. SF samples were obtained by puncture after scleral indentation. TBARS and protein concentrations were determined by the corresponding colorimetric assays. A linear correlation exists between TBARS and protein contents in these samples. No correlation could be established between evolution time of the retinal detachment and TBARS content in SF. TBARS in SF increases with increasing age in nonmyopic patients. In the samples of myopic patients the correlation was established between TBARS content and degree of myopia. The group of patients with more than 10 dioptres show a significant higher TBARS concentration in SF than any of the other groups studied. It can be concluded that lipid peroxidation products in SF originate, at least partially, from rod outer segments, and that lipid peroxidation is a process that might play a role in the pathogenesis of retinal detachment, specially in myopic patients.     

Egg yolk improves lipid profile, lipid peroxidation and retinal abnormalities in a murine model of genetic hypercholesterolemia

Carotenoids are believed to inhibit oxidative stress. We investigated the protective effect of lutein and egg yolk supplementation on systemic and retinal alterations in apolipoprotein E-deficient (apoE-/-) mice, an experimental model of hypercholesterolemia and cardiovascular disease. Three-month-old wild-type and apoE-/- mice received one of the following: vehicle, lutein (0.09 mg/kg per day) or egg yolk (0.8 g/kg per day), by gastroesophageal cannula for 3 months. Total cholesterol (TC), triacylglycerol (TG) and lipid peroxidation (TBARS) were measured in plasma. TBARS levels were also determined in retinal homogenates. Ultrastructural morphology was analyzed by electron microscopy. ApoE-/- mice, with increased TC and TG concentrations, had higher systemic (P<.05) and retinal (P<.01) levels of lipid peroxidation than wild-type strains. Electron microscopy showed ultrastructural alterations (basal laminar deposits, open intercellular junctions, increased cytoplasmic vacuoles) in the retinas from apoE-/- mice. Egg yolk significantly reduced plasma TG (P<.05) and, without changes in TC, decreased plasma lipid peroxidation (P<.05). Lutein supplementation marginally affected the parameters. Less severe retinal ultrastructural alterations were observed in apoE-/- mice receiving either egg yolk or lutein. In the apoE-/- mouse model, egg yolk improved the lipid profile and reduced systemic lipid peroxidation (P<.05). While lutein and egg yolk did not seem to reduce retinal lipid peroxidation, a reduction in retinal ultrastructural alterations was observed.     

Immunohistochemical distribution of basic fibroblast growth factor in experimental retinal ischaemia and reperfusion in the rat

Summary It has been proposed that basic fibroblast growth factor (basic FGF) mediates the neovascular response in a variety of conditions, including diabetic retinopathy and branch retinal vein occlusion. To test the hypothesis that basic FGF was released from retinal stores as a result of retinal ischaemia, transient retinal ischaemia was induced, followed by 48 h of reperfusion, in the rat by combined central retinal vasculature and optic nerve ligation. The immunolocalization of basic FGF was studied in the retina. We found that basic FGF in the normal retina is present around the deeper retinal vessels and in the neuronal tissue of the outer plexiform layer. In the eyes that had ischaemia followed by reperfusion, there was moderate cellular oedema with retinal swelling, and mitoses in the inner nuclear and plexiform layers. There were no changes evident at the immunohistochemical level either in the intensity or distribution of stores of basic FGF. We conclude from these data that stores of basic FGF are not altered dramatically under the conditions of transient experimental ischaemia and reperfusion in the rat, despite the presence of cellular proliferation.     

米诺环素对糖尿病大鼠视网膜神经细胞凋亡的影响

目的：观察米诺环素对糖尿病大鼠视网膜神经细胞的凋亡的影响,研究米诺环素对糖尿病视网膜神经保护作用,对米诺环素在糖尿病视网膜疾病中抑制神经细胞凋亡提供理论支持。方法：选择健康成年雄性SD大鼠30只,随机分成正常对照组、糖尿病模型组和米诺环素治疗组,每组10只。腹腔内注射链脲佐菌素（STZ）诱发大鼠糖尿病。米诺环素治疗纽给予米诺环素腹腔注射（45mg／kg）,共注射10d,模型组和对照组腹腔注射等体积生理盐水,于给药后8周的3组动物处死,冰浴下取其视网膜组织,随后制备视网膜石蜡切片,采用末端脱氧核糖核酸介导生物素化脱氧尿嘧啶缺口末端标记L（TUNEL）法进行凋亡细胞原位标记,行视网膜神经细胞凋亡计数,对所有数据进行统计学分析。实验结果拟用均数和标准差（x±s）表示,P〈0．05为差异有统计学意义。结果 相同观察时相,与阴性对照组比较,模型对照组大鼠视网膜TUNEL阳性细胞显著增多（P〈0．01）,在米诺环素组,大鼠视网膜TUNEL阳性细胞数比模型对照组明显减少,差异有统计学意义（P〈0．01）。结论：米诺环素能有效降低糖尿病大鼠视网膜神经细胞的凋亡,对糖尿病视网膜神经细胞有保护作用。     

Changes in the function and ultrastructure of vessels in the rat model of multiple low dose streptozotocin-induced diabetes

In this study we investigated functional changes in the femoral artery and ultrastructural alterations in mesenteric vessels and capillaries in the rat model of multiple low dose streptozotocin (STZ)-induced diabetes. Participation of oxidative stress in this model of diabetes was established by studying the effect of the pyridoindole antioxidant stobadine (STB) on diabetes-induced impairment. Experimental diabetes was induced by i.v. bolus of STZ (20 mg/kg) given for three consecutive days to male rats. At the 12(th) week following STZ administration, the animals revealed typical signs of diabetes, such as polyphagia, polydypsia and polyuria. There was no weight gain in the diabetic groups throughout the experiment. No exitus occurred in any group. Diabetes was characterised with high levels of plasma glucose, no significant changes in lipid metabolism, decreased serum levels of glutathione, increased serum levels of the lysosomal enzyme N-acetyl-beta-D-glucosaminidase (NAGA), injured endothelial relaxant capacity of the femoral artery and alterations in ultrastructure of mesenteric arteries and capillaries. Antioxidant STB in the dose of 25 mg/kg body weight i.p. (5 times per week) did not influence glucose levels, however, it mitigated biochemical, functional and ultrastructural changes induced by diabetes, suggesting a role of reactive oxygen species in diabetes-induced tissue damage.     

Strain-Independent Increases of Crystallin Proteins in the Retina of Type 1 Diabetic Rats

Diabetic retinopathy is the leading cause of vision loss in working-age individuals in the United States and is expected to continue growing with the increased prevalence of diabetes. Streptozotocin-induced hyperglycemia in rats is the most commonly used model for diabetic retinopathy. Previous studies have shown that this model can lead to different inflammatory changes in the retina depending on the strain of rat. Our previous work has shown that crystallin proteins, including members of the alpha- and beta/gamma-crystallin subfamilies, are upregulated in the STZ rat retina. Crystallin proteins have been implicated in a number of cellular processes, such as neuroprotection, non-native protein folding and vascular remodeling. In this current study, we have demonstrated that unlike other strain-dependent changes, such as inflammatory cytokines and growth factor levels, in the STZ rat, the protein upregulation of crystallins is consistent across the Brown Norway, Long-Evans and Sprague-Dawley rat strains in the context of diabetes. Taken together, these data illustrate the potential critical role played by crystallins, and especially alpha-crystallins, in the retina in the context of diabetes.