Interaction between the polyol pathway and non-enzymatic glycation on aortic smooth muscle cell migration and monocyte adhesion

We investigated for the interaction between the polyol pathway and enhanced non-enzymatic glycation, both implicated in the pathogenesis of diabetic atherosclerosis, in the activation of aortic smooth muscle cell (SMC) function. Mouse aortas and primary cultures of SMCs from wildtype (WT) mice and transgenic (TG) mice expressing human aldose reductase (AR) were studied regarding changes in AR activity, and SMC gene activation, migration and monocyte adhesion, in response to advanced glycation end-product modified BSA (AGE-BSA). Results showed that AGE-BSA increased AR activity in both WT and TG aortas, with greater increments (p < 0.05) in TG aortas which, basally, had elevated AR activity (2.8 fold of WT). These increments were attenuated by zopolrestat, an AR inhibitor. Similar AGE-induced increments in AR activity were observed in primary cultures of aortic SMCs from WT and TG mice (60% and 100%, respectively, P < 0.01). Such increments were accompanied by increases in intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) mRNA levels (both P < 0.05), activation of membrane-associated PKC-beta1 (P < 0.05) as well as increased SMC migration and Tamm-Horsfall protein (THP)-1 monocyte adhesion to SMCs (both p < 0.01), with all changes being significantly greater in TG SMCs (P < 0.05) and suppressible by either zopolrestat or transfection with an AR antisense oligonucleotide. Our findings suggest that the effects of AGEs on SMC activation, migration and monocyte adhesion are mediated partly through the polyol pathway and, possibly, PKC activation. The greater AGE-induced changes in the TG SMCs have provided further support for the dependency of such changes on polyol pathway hyperactivity.     

Cigarette smoke concentrate increases 8-epi-PGF2alpha and TGFbeta1 secretion in rat mesangial cells

Epidemiological studies have shown that cigarette smoke, an oxidant agent, is a risk factor for the development of diabetic nephropathy (DN), in which pathogenesis transforming growth factor beta(1) (TGFbeta(1)) plays a key role. In our experimental model we exposed mesangial cell cultures to cigarette smoke concentrate (CSC) to study the effect of smoking on the pathogenesis of DN. Thus, we analyzed the effect of CSC on TGFbeta(1) and lipid peroxidation (8-epi-PGF(2alpha)) in rat mesangial cells. Furthermore, since the protein kinase C (PKC) pathway appears to be a key factor for the enhanced production of TGFbeta(1), we also analyzed the effect of the selective PKCbeta inhibitor LY379196 on TGFbeta(1) response to CSC. CSC induced an increase of both TGFbeta(1) and 8-epi-PGF(2) compared to basal conditions (5 mM glucose). The CSC-induced increase in TGFbeta(1) secretion was significantly suppressed by LY379196. These data suggest that smoking could increase TGFbeta(1) production, probably due to oxidative stress and PKCbeta activation. This finding supports the concept that smoking is a risk factor for DN development.     

延长糖尿病模型大鼠生存期对糖尿病视网膜病变的影响

目的延长糖尿病模型大鼠生存期,动态观察糖尿病视网膜病变(DR)的形成和发展过程。方法雄性SD大鼠70只,随机分成对照组(20只)和模型组(50只),采用链脲佐菌素(STZ)60 mg/(kg.bw)体重腹腔1次注射造模,分别于69、、12月时处死取眼球,采用视网膜微血管消化铺片技术观察糖尿病视网膜病变的微血管形态学改变。结果糖尿病大鼠DR样病变随着病程的延长病变呈多样性改变,以12月DR出现的小动脉硬化尤为严重。结论糖尿病大鼠生存期的延长对糖尿病视网膜病变的研究有着积极的意义。     

糖尿病视网膜病变的基因治疗

糖尿病视网膜病变（diabetic retinopathy,DR）是糖尿病的主要并发症之一,大量的研究表明DR早期表现为视网膜神经细胞退化性改变,晚期出现新生血管及增殖膜,目前尚没有有效的治疗方法,严重危害患者视力。基因治疗为探索DR的治疗方法提供了新的手段,本文就目前DR的基因治疗研究现状和进展作一综述。     

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。     

C 肤对糖尿病大鼠坐骨神经结构和功能的影响

目的：探讨外源性C肽对Ⅰ型糖尿病大鼠坐骨神经结构及功能的影响。方法：选取Wistar大鼠40只,分为正常对照组（NC组）和糖尿病组（Dia组）,糖尿病组链脲佐菌素诱发大鼠成模后,再随机分为三组：糖尿病组（Dia组）、胰岛素治疗组（In组）和C肽治疗组（CP组）8周后,测定各组大鼠运动、感觉神经传导速度,并对病变大鼠的坐骨神经进行病理定量图像分析及超微结构分析,结果：1．In组、CP肽组与DM组相比：大鼠运动、感觉神经传导速度均明显增加（P〈0．01）;2．腓肠神经纤维的数量和总横截面面积也显著增加（P〈0.01）。3．CP组与In组相比运动、感觉神经传导速度也显著增加（P〈0．01）。4．电镜显示：Dia组有髓神经纤维髓鞘发生分离并有无颗粒囊胞状结构聚集现象,In组有髓神经纤维髓鞘分离现象明显减轻但仍有无颗粒囊胞状结构聚集现象．而CP组有髓神经纤维结构完全接近正常组。结论：C肽在改善糖尿病大鼠的神经结构和功能方面明显优于胰岛素．     

大鼠半乳糖性白内障动物模型制备方法的改良及其机制

目的改良大鼠半乳糖性白内障动物模型制备方法 ,比较改良前后两种方法各自的特点及其机制。方法 60只体重（180±10）g的雄性成年SD大鼠随机分为空白对照组（C）、半乳糖性白内障组1（G1）和半乳糖性白内障组2（G2）。G1组腹腔注射50%半乳糖溶液20mL/（kg·d）,连续30d;G2组第1周、第2周分别腹腔注射50%半乳糖溶液10、15mL/（kg·d）,第3周开始增至20mL/（kg·d）,直至第30d。应用裂隙灯观察晶体混浊情况,按大鼠晶状体混浊度评分标准进行分级和记录,并分别观察和比较不同制备方法的成模时间、模型成功率、模型死亡率和晶状体宏观形态变化以及SOD、MDA含量的变化。结果 G1和G2组模型死亡率分别为50%和10%,方法改良后死亡率显著降低;模型成功率分别为50%和90%,方法改良后成功率显著提高;两组模型晶状体混浊度一般为Ⅱ级或Ⅲ级;与C组比较,G1和G2组SOD、MDA含量的变化均有极显著差异（P﹤0.01）,而G2与G1组比较,差异不显著。结论半乳糖性白内障动物模型制备方法改良后,保留了改良前模型制备方法的优点,克服了改良前模型成活率低、死亡率高的缺点,是一种更为安全、可靠的模型制备方法 。     

链脲佐菌素诱导长爪沙鼠Ⅰ型糖尿病模型的实验研究

目的探讨链脲佐菌素(STZ)诱导长爪沙鼠Ⅰ型糖尿病模型的可能性,并观察模型动物早期肾脏损害情况。方法雄性长爪沙鼠96只,随机分为正常对照组(NC组)、模型组1(DM1组)、模型组2(DM2组),DM1及DM2组沙鼠分别一次性腹腔注射100 mg/kg、200 mg/kg STZ,NC组注射等量柠檬酸盐缓冲溶液。注射STZ后1、2、4、6周末,分别监测沙鼠一般情况,血糖、胰岛素等血清学指标和尿液指标,并处死沙鼠进行胰腺和肾脏组织的病理学检查。结果注射STZ 24 h后,DM2组及DM1组部分沙鼠逐渐出现典型的"三多一少"症状,随着病程的发展,DM2组沙鼠持续高血糖,DM1组沙鼠血糖值与NC组差异有显著性(P0.05),但有下降趋势;DM2组沙鼠胰岛素显著性降低(P0.05),其他血清学指标及尿液指标均显著性升高(P0.05),DM1组沙鼠各指标差异无显著性。DM2组沙鼠及DM1组少数沙鼠胰腺组织中可见胰岛β细胞减少、空泡样变性等变化,DM2组沙鼠肾脏组织中出现肾小球基质增多,毛细血管襻扩张等病变,DM1组沙鼠肾脏组织未见明显变化。结论 STZ 200 mg/kg可成功诱导长爪沙鼠Ⅰ型糖尿病模型,在病程早期沙鼠肾脏结构和功能已经发生改变。     

Lens‐anomalies and other ophthalmic findings in a group of closely‐related angola lions (Panthera leo bleyenberghi)

After the diagnosis of bilateral, immature, nuclear, and posterior cortical cataracts in one Angola lioness, and because of the possible implications of the cataracts for a breeding program, complete ophthalmic examinations on a group of related adult Angola lions and their offspring were carried out. Five adult lions, ranging in age from 1.5–5.5 years, and five lion cubs were studied clinically. The examination included slit‐lamp biomicroscopy, indirect ophthalmoscopy, and photography. The eyes of three of the offspring were submitted for histopathologic examination and examined by light microscopy. The most significant findings were cataracts of various stages, which were observed in four adult lions and one male cub. Mild lenticular abnormalities were noted in the histopathologic examination of the lion cubs' eyes. Additional ophthalmic findings, of lesser clinical consequence, were also noted. This breeding program would benefit from further investigation by animal nutritionists and geneticists, and the animals in this group should undergo periodic ophthalmologic examinations. Zoo Biol 0:1–7, 2006. © 2006 Wiley‐Liss, Inc.     

Upregulation of Glyoxalase I fails to Normalize Methylglyoxal Levels: A Possible Mechanism for Biochemical Changes in Diabetic Mouse Lenses

Glyoxalase I is the first enzyme in a two-enzyme glyoxalase system that metabolizes physiological methylglyoxal (MGO). MGO reacts with proteins to form irreversible adducts that may lead to crosslinking and aggregation of lens proteins in diabetes. This study examined the effect of hyperglycemia on glyoxalase I activity and its mRNA content in mouse lens epithelial cells (mLE cells) and in diabetic mouse lenses and investigated the relationship between GSH and MGO in organ cultured lenses. mLE cells cultured with 25 mM D-glucose (high glucose) showed an upregulation of glyoxalase I activity and a higher content of glyoxalase I mRNA when compared with either cells cultured with 5 mM glucose (control) or with 20 mM L-glucose + 5 mM D-glucose. MGO concentration was significantly elevated in cells cultured with high D-glucose, but not in L-glucose. GSH levels were lower in cells incubated with high glucose compared to control cells. Glyoxalase I activity and mRNA levels were elevated in diabetic lenses compared to non-diabetic control mouse lenses. MGO levels in diabetic lenses were higher than in control lenses. Incubation of lenses with buthionine sulfoximine (BSO) resulted in a dramatic decline in GSH but the MGO levels were similar to lenses incubated without BSO. Our data suggest that in mouse lenses MGO accumulation may occur independent of GSH concentration and in diabetes there is an upregulation of glyoxalase I, but this upregulation is inadequate to normalize MGO levels, which could lead to MGO retention and chemical modification of proteins.