糖尿病大鼠阴茎海绵体GSH、GSSG和GSH/GSSG对eNOS表达的影响

目的探究在糖尿病大鼠阴茎海绵体中还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)和GSH/GSSG对内皮型一氧化氮合酶(eNOS)表达的影响。方法 30只SD级8周龄健康雄性大鼠,8只作为正常对照组(C组),剩余22只作为糖尿病实验组(D组);D组大鼠高脂高糖喂养4周后腹腔注射链脲佐菌素(40mg/kg)建DM模型;C组大鼠未作特殊处理。继续喂养8周后,注射阿扑吗啡(apomorphine,APO),观察阴茎勃起情况;所有大鼠测定阴茎海绵体内压/平均颈动脉压(ICP_(max)/MAP)后处死,采集血液、阴茎组织,测定GSH、GSSG、睾酮含量,免疫组化和Western印迹法检测eNOS的表达。结果与C组相比较,D组大鼠阴茎组织中GSH含量、GSH/GSSG比值、eNOS含量明显降低(P0.05),而GSSG的含量明显升高(P0.05)。结论糖尿病大鼠阴茎海绵体中GSH和GSSG、GSH/GSSG影响eNOS的表达,在糖尿病性阴茎勃起功能障碍机制中起重要作用。     

负压吸引对糖尿病性ED大鼠阴茎海绵体组织一氧化氮合酶表达的影响

目的研究负压吸引对糖尿病性勃起功能障碍（ED）大鼠阴茎组织一氧化氮合酶（NOS）表达水平的影响。方法25只实验鼠中随机选取5只为正常对照组（A组）,其余火鼠用链脲左菌素和阿朴吗啡诱导建立Ⅰ型糖尿病性ED大鼠模型。之后把造模成功的糖尿病性ED大鼠随机分成糖尿病ED吸引组（B组）和糖尿病ED非吸引组（C组）。在B组大鼠负压吸引治疗结束后将A、B、C3组大鼠处死并取阴茎组织进行石蜡包埋。采用免疫组织化学方法检测各组大鼠阴茎组织中三种一氧化氮合酶亚型（nNOS、eNOS、iNOS）的表达情况。结果A组大鼠阴茎组织中nNOS蛋白表达水平高于B组和C组（均P〈0.001）;A组和B组大鼠阴茎组织中eNOS蛋白表达水平高于C组（均P〈0.01）;A组iNOS蛋白表达水平低于B组和C组（P〈0.01,P〈0.001）,同时B组iNOS蛋白表达水平低于C组（P〈0.01）;剩余其他各组间的比较差异无统计学意义（P〉0.05）。结论负压吸引可以通过升高阴茎组织中的eNOS和降低iNOS的表达来改善勃起功能。     

α-硫辛酸通过逆转内皮型一氧化氮合成酶过表达改善糖尿病膀胱功能

目的 明确抗氧化治疗方法 对糖尿病膀胱病变的治疗效果,探讨该方法 的作用机制.方法 制备链脲佐菌素(STZ)-糖尿病大鼠模型;通过尿流动力学检查方法 明确正常对照组、糖尿病组和治疗组的膀胱功能状态,观察α-LA对大鼠排尿功能的治疗效果;应用逆转录-聚合酶链反应(RT-PCR)和Western blot方法 检测α-LA治疗前后各组大鼠膀胱组织中内皮型一氧化氮合成酶(eNOS)及其代谢标志物3-硝基酪氨酸(3-NT)的表达水平.结果 糖尿病大鼠的最大膀胱容量、单次排尿量、残余尿量均呈时间依赖性显著增加(P＜0.01);膀胱排尿率亦显著下降(P＜0.05),经仅α-LA治疗后,上述指标显著改善.α-LA治疗后,糖尿病大鼠膀胱组织中显著增加的eNOS、3-NT表达亦明显下降.结论 α-LA可能通过抑制eNOS过表达减少氧化应激、改善糖尿病膀胱病变.     

糖尿病大鼠阴茎海绵体组织中一氧化氮合酶亚型表达的研究

目的:通过观察糖尿病性雄性大鼠阴茎海绵体组织中一氧化氮合酶亚型(nNOS、iNOS、eNOS)表达和勃起功能的变化,以及应用胰岛素、α-硫辛酸(LA)干预对其的影响,进一步探讨糖尿病性ED发病机制,并为临床治疗提供新的思路。方法:将50只成年雄性SD大鼠分为4组:A组为正常对照组(10只),B组为糖尿病无干预组(13只),C组为糖尿病胰岛素干预组(12只),D组为糖尿病胰岛素+LA干预组(15只),采用腹腔注射链脲佐菌素法制作糖尿病模型。8周后各组大鼠通过注射阿朴吗啡后评价勃起功能并取阴茎海绵体组织用免疫组化EnV i-sion法观察海绵体组织中nNOS、iNOS、eNOS表达的变化。结果:A组大鼠勃起功能正常,勃起率100%;与A组相比,B组、C组、D组勃起功能水平显著降低(P<0.05),勃起率:B组28.6%,C组62.5%,D组80.9%,其海绵体组织中nNOS、eNOS表达显著降低,A组大鼠海绵体组织每视野nNOS、eNOS阳性神经纤维数为86.7、9.6,B组为36.5、3.3,C组为52.7、5.7,D组为71.4、7.4,而iNOS表达显著增加(P<0.05),A组为6.9,B组为43.6,C组为36.2,D组为19.3;与B、C组相比,D组勃起功能显著上升,海绵体组织中nNOS、eNOS表达均显著上升,iNOS显著下降(P<0.05)。结论:糖尿病严重影响阴茎勃起功能和nNOS、iNOS、eNOS的表达,高血糖是其发病基础之一,而LA对其有显著疗效,推测可能与其抗氧化作用机制有关。     

伊木萨克片对糖尿病性ED大鼠阴茎组织中eNoS和nNoS的影响

目的 研究维药伊木萨克片对DM性ED大鼠阴茎海绵体组织中eNOS和nNOS蛋白表达水平的影响.方法 取雄性SD大鼠70只,从中随机取10只为正常对照组,余60只以链脲佐菌素诱导建立DM模型后,行阿朴吗啡阴茎勃起实验筛选DM性ED模型,发生ED者随机分为DM性ED组、伊木萨克片组、胰岛素组、伊木萨克片 胰岛素(联用)组,未成DM者为STZ组.各组给药6周后,采用免疫组化SP法检测各组大鼠阴茎组织中eNOS、nNOS含量.结果 DM性ED组大鼠阴茎组织中eNOS、nNOS蛋白表达水平显著低于正常对照组(P＜0.01);伊木萨克片组、胰岛素组及联用组大鼠阴茎组织中eNOS、nNOS蛋白表达水平均显著高于DM性ED组(P＜0.01),但伊木萨克片组与胰岛素组显著低于联用组(P＜0.01, P＜0.01;P＜0.05,P＜0.01);STZ组与正常对照组之间的差异则无统计学意义(P＞0.05).结论 维药伊木萨克片可以显著提高DM性ED大鼠阴茎组织中eNOS、nNOS蛋白表达水平,并提示在胰岛素控制血糖的基础上效果可能更佳.     

枸杞多糖对糖尿病大鼠阴茎海绵体氧化应激损伤的保护作用

目的探讨枸杞多糖(LBP)对糖尿病大鼠阴茎海绵体氧化应激损伤的保护作用。方法 SD大鼠腹腔注射链脲佐菌素(STZ)建立糖尿病大鼠模型,随机设立糖尿病组(DM组)、枸杞多糖灌胃组(LBP组),并设立正常对照组(NC组)。8周后通过APO试验检测勃起功能,硫代巴比妥酸法检测阴茎海绵体组织内丙二醛(MDA)含量,黄嘌呤氧化酶法检测阴茎海绵体组织内超氧化物歧化酶(SOD)含量,光镜下观察阴茎海绵体组织Masson染色病理切片中胶原纤维、平滑肌纤维的差异。结果与NC组比较,DM组大鼠勃起次数明显减少(P0.05),阴茎海绵体组织内MDA含量显著增高(P0.05),SOD含量明显减少(P0.01);大鼠阴茎海绵体组织平滑肌部分断裂,数目减少,胶原纤维菲薄,结构破坏,排列紊乱。经灌胃8周后,LBP组与DM组比较:勃起次数增多(P0.05),大鼠阴茎海绵体组织中MDA含量降低(P0.05),SOD含量明显增加(P0.01),光镜下观察阴茎海绵体组织结构改善。结论实验证实,LBP可以减轻糖尿病大鼠阴茎海绵体氧化应激损伤,修复部分组织结构,改善大鼠勃起功能。     

左旋精氨酸对糖尿病大鼠阴茎海绵体氧化应激损伤的保护作用

目的 探讨左旋精氨酸(L-Arg)对糖尿病大鼠阴茎海绵体氧化应激损伤的保护作用.方法 大剂量STZ腹腔注射建立糖尿病大鼠模型,随机分为糖尿病组、L-Arg治疗组,并设正常对照组.治疗8周后用电刺激各组大鼠勃起神经测定海绵体内压评价勃起功能采用黄嘌呤氧化酶法检测阴茎海绵体组织中超氧化物歧化酶(SOD)活性,硫代芭比妥酸法测丙二醛(MDA)含量;光镜下观察Masson染色切片.结果 糖尿病组较正常对照组,阴茎海绵体MDA含量显著增高(P<0.01),SOD活性显著下降(P<0.05),最大海绵体内压(ICP)显著降低(P<0.05);与糖尿病组比较,L-Arg可使海绵体MDA含量明显降低(P<0.05),显著提高其SOD活性及ICP(P<0.05).糖尿组大鼠阴茎组织中胶原纤维含量明显减少,排列紊乱、稀疏,平滑肌减少变薄,出现断裂;L-Arg组大鼠阴茎组织结构可见明显改善.结论 糖尿病大鼠阴茎海绵体存在氧化应激损伤,导致海绵体结构的改变是其勃起水平下降的重要因素,通过补充L-Arg,可以减轻勃起组织的氧化应激损伤,改善组织结构,从而提高勃起能力.     

白藜芦醇对糖尿病心肌病小鼠心肌细胞铁死亡的影响

目的评价白藜芦醇对糖尿病心肌病小鼠心肌细胞铁死亡的影响。方法清洁级健康成年雄性C57BL/6小鼠30只, 8周龄, 体质量22～26 g。采用随机数字表法将其分为3组(n=10):对照组(C组)、糖尿病心肌病组(DCM组)和白藜芦醇组(RSV组)。采用连续5 d腹腔注射新鲜制备的链脲佐菌素40 mg·kg-1·d-1的方法制备小鼠糖尿病心肌病模型。模型制备成功后, RSV组小鼠连续12周经灌胃给予白藜芦醇25 mg·kg-1·d-1, C组和DCM组小鼠给予等体积二甲基亚砜。于第12周末行超声心动图检查心脏结构及心功能情况, 随后处死小鼠收集心肌组织标本, 采用HE染色观察小鼠心肌组织病理学结果, 透射电镜下观察小鼠心肌组织线粒体结构, 采用比色法测定铁、MDA和谷胱甘肽(GSH)含量, Western blot法检测谷胱甘肽还原酶4(GPX4)表达。结果与C组比较, DCM组小鼠左室舒张末期内径(LVDd)和左室收缩末期内径(LVDs)升高, 左室短轴缩短率(LVFS)和左室射血分数(LVEF)降低, 心肌组织铁和MDA含量升高, GSH含量降低, GPX4表达下调(P<0.0...     

白藜芦醇联合二苯乙烯苷通过抗衰老延缓糖尿病肾病进展的研究

目的:探讨白藜芦醇及二苯乙烯苷联合干预对糖尿病肾病进展的影响及其可能机制。方法:本实验通过腹腔注射链脲佐菌素诱导大鼠糖尿病模型,白藜芦醇联合二苯乙烯苷灌胃干预24周,收集血、尿、肾组织标本行肾功能、α-Klotho蛋白、氧化应激等检测。结果:白藜芦醇联合二苯乙烯苷治疗可显著降低糖尿病肾病大鼠尿白蛋白、尿蛋白、血尿素氮及肌酐水平,并且明显降低尿液NGAL、KIM-1水平。通过检测与衰老相关的α-Klotho及氧化应激相关指标发现,上述治疗作用与下调肾脏ROS、MDA含量、抑制尿液8-OHdG及维持血液α-Klotho含量密切相关。结论:白藜芦醇联合二苯乙烯苷可能通过对抗肾脏衰老延缓糖尿病肾病进展。     

丙戊茶碱对糖尿病痛性周围神经病变大鼠脊髓星形胶质细胞活化的影响

目的 评价丙戊茶碱对糖尿病痛性周围神经病变大鼠脊髓星形胶质细胞活化的影响.方法 清洁级SD大鼠40只,月龄2月,雌雄不拘,体重180-200 g,腹腔注射1%链脲佐菌素60 mg/kg以制备大鼠糖尿病痛性周围神经病变模型.取模型制备成功的大鼠20只,随机分为糖尿病组(DM组)和丙戊茶碱组(PP组),每组10只;另取10只同月龄D大鼠作为对照组(NC组).PP组和DM组在注射链脲佐菌素后第28天开始,每天分别腹腔注射丙戊茶碱10 mg/kg和等容量生理盐水,注射1周.于注射链脲佐菌素前(TI)、注射链脲佐菌素后2、7、14、21、28、35 d(T2-7)时取尾静脉血样0.1 ml测定血糖水平;于T1、T3-7,时测定大鼠右后爪机械缩足反应阈值(PWT);T7时处死大鼠,取L4,s脊髓制备切片.采用免疫组化法检测神经胶质纤维酸性蛋白(GFAP)表达.结果 与TI时比较,DM组和PP组L2-7时血糖水平升高,T4-7时PWT降低(P<0.01);与NC组比较,DM组和PP组T2-7时血糖水平升高,T6,7时PWT降低,T7时GFAP表达上调(P<0.01);与DM组比较,T7时PWT升高,GFAP表达下调(P<0.05).结论 丙戊茶碱可能通过抑制脊髓星形胶质细胞活化减轻大鼠糖尿病神经病理性痛.     

一氧化氮在人参皂甙Rb1预处理减轻糖尿病大鼠心肌缺血再灌注损伤中的作用

目的 评价一氧化氮(NO)在人参皂甙Rb1预处理减轻糖尿病大鼠心肌缺血再灌注损伤中的作用.方法 成年雄性SD大鼠,体重220～280g,腹腔注射1%链脲佐菌素-柠檬酸盐缓冲液65 mg/kg制备糖尿病模型.取糖尿病模型制备成功的大鼠40只,随机分为4组(n=10):假手术组(S组)、缺血再灌注组(IR组)、人参皂甙Rb1预处理组(R组)和L-NAME+人参皂甙Rb1预处理组(LR组).IR组、R组和LR组采用结扎左冠状动脉前降支30 min,再灌注120 min的方法制备大鼠心肌缺血再灌注模型;S组只穿线.LR组于缺血前25 min时静脉注射一氧化氮合酶抑制剂N-硝基-L-精氨酸甲酯10 mg/kg;R组和LR组于缺血前10 min时静脉注射人参皂甙Rb1 40mg/kg;S组和IR组给予等容量生理盐水.再灌注120 min时,颈动脉采集血样,测定血清肌酸激酶(CK)和乳酸脱氢酶(LDH)的活性.然后处死大鼠,取心肌组织,计算心肌梗死范围,测定内皮型一氧化氮合酶(eNOS)表达、MDA和NO的含量以及SOD活性,光镜下观察病理结果.结果 与S组比较,IR组、R组和LR组血清CK和LDH的活性升高,心肌梗死范围增大,IR组和LR组心肌eNOS表达下调,MDA含量升高,SOD活性和NO含量降低(P＜0.05);与IR组和LR组比较,R组血清CK和LDH的活性降低,心肌梗死范围减小,心肌eNOS表达上调,MDA含量降低,SOD活性和NO含量升高(P＜0.05);IR组和LR组各指标比较差异无统计学意义(P＞0.05).结论人参皂甙Rb1预处理可通过激活eNOS,促进NO生成,抑制心肌细胞脂质过氧化反应,从而减轻糖尿病大鼠心肌缺血再灌注损伤.     

Hyperglycemia-induced apoptosis in mouse myocardium: mitochondrial cytochrome C-mediated caspase-3 activation pathway

Diabetic cardiomyopathy is related directly to hyperglycemia. Cell death such as apoptosis plays a critical role in cardiac pathogenesis. Whether hyperglycemia induces myocardial apoptosis, leading to diabetic cardiomyopathy, remains unclear. We tested the hypothesis that apoptotic cell death occurs in the diabetic myocardium through mitochondrial cytochrome c-mediated caspase-3 activation pathway. Diabetic mice produced by streptozotocin and H9c2 cardiac myoblast cells exposed to high levels of glucose were used. In the hearts of diabetic mice, apoptotic cell death occurred as detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Correspondingly, caspase-3 activation as determined by enzymatic assay and mitochondrial cytochrome c release detected by Western blotting analysis were observed. Supplementation of insulin inhibited diabetes-induced myocardial apoptosis as well as suppressed hyperglycemia. To explore whether apoptosis in diabetic hearts is related directly to hyperglycemia, we exposed cardiac myoblast H9c2 cells to high levels of glucose (22 and 33 mmol/l) in cultures. Apoptotic cell death was detected by TUNEL assay and DAPI nuclear staining. Caspase-3 activation with a concomitant mitochondrial cytochrome c release was also observed. Apoptosis or activation of caspase-3 was not observed in the cultures exposed to the same concentrations of mannitol. Inhibition of caspase-3 with a specific inhibitor, Ac-DEVD-cmk, suppressed apoptosis induced by high levels of glucose. In addition, reactive oxygen species (ROS) generation was detected in the cells exposed to high levels of glucose. These results suggest that hyperglycemia directly induces apoptotic cell death in the myocardium in vivo. Hyperglycemia-induced myocardial apoptosis is mediated, at least in part, by activation of the cytochrome c-activated caspase-3 pathway, which may be triggered by ROS derived from high levels of glucose.     

Role of endothelin receptors and relationship with nitric oxide synthase in impaired erectile response in diabetic rats

To evaluate the protective role of bosentan (BOS), an endothelin‐1 (ET‐1) receptor antagonist, and to show the changes in rats with experimentally induced diabetic erectile dysfunction (ED), a total of 24 albino Wistar rats were allocated into four groups. Group 1 was the healthy group and Group 2 had diabetes mellitus (DM) induced by intraperitoneal injection of 60 mg kg^?1 streptozotocin (STZ). Following the establishment of DM, Group 3 and Group 4 were treated with oral BOS doses of 50 mg kg^?1 and 100 mg kg^?1, respectively, for 60 days. At the end of the treatment, we evaluated yawning and erection response to apomorphine treatment and then the animals were sacrificed. ET‐1, eNOS, iNOS, tumour necrosis factor (TNF)‐α, ET‐RA and ET‐RB mRNA expressions were analysed in cavernosal tissue. It was observed that yawning and erection response decreased in the diabetic group; however, both of these improved with BOS treatment. While ET‐1, TNF‐α and iNOS gene expressions increased, eNOS, ET‐RA and ET‐RB gene expressions decreased in the DM group compared to the healthy group. DM has a negative impact on cavernosal tissue blood flow through activating vasoconstrictor mediators in cavernosal tissue. BOS regulates significantly eNOS, iNOS and TNF‐α expressions in a dose‐dependent manner.     

Transplantation of endothelial progenitor cells transfected with VEGF165 to restore erectile function in diabetic rats

The present study investigated the effect of transplanting endothelial progenitor cells (EPCs) transfected with the vascular endothelial growth factor gene (VEGF165) into the corpora cavernosa of rats with diabetic erectile dysfunction (ED). A rat model of diabetic ED was constructed via intraperitoneal injection of streptozotocin. After streptozotocin treatment, pre-treated EPCs from each of three groups of rats were transplanted into their corpora cavernosa. Our results, following intracavernosal pressure (ICP) monitoring, showed that ICP increased significantly among rats in the trial group when compared to the results from rats in the blank-plasmid and control groups during basal conditions and electrical stimulation (P<0.01 for both comparisons). Histological examination revealed extensive neovascularisation in the corpora cavernosa of rats in the trial group. Fluorescence microscopy indicated that many of the transplanted EPCs in the trial group survived, differentiated into endothelial cells and integrated into the sites of neovascularisation. Based on the results of this study, we conclude that transplantation of VEGF165-transfected EPCs into the corpora cavernosa of rats with diabetic ED restores erectile function.     

Endothelial nitric oxide synthase uncoupling impairs endothelial progenitor cell mobilization and function in diabetes

Uncoupling of the endothelial nitric oxide synthase (eNOS) resulting in superoxide anion (O(2)(-)) formation instead of nitric oxide (NO) causes diabetic endothelial dysfunction. eNOS regulates mobilization and function of endothelial progenitor cells (EPCs), key regulators of vascular repair. We postulate a role of eNOS uncoupling for reduced number and function of EPC in diabetes. EPC levels in diabetic patients were significantly reduced compared with those of control subjects. EPCs from diabetic patients produced excessive O(2)(-) and showed impaired migratory capacity compared with nondiabetic control subjects. NOS inhibition with N(G)-nitro-l-arginine attenuated O(2)(-) production and normalized functional capacity of EPCs from diabetic patients. Glucose-mediated EPC dysfunction was protein kinase C dependent, associated with reduced intracellular BH(4) (tetrahydrobiopterin) concentrations, and reversible after exogenous BH(4) treatment. Activation of NADPH oxidases played an additional but minor role in glucose-mediated EPC dysfunction. In rats with streptozotocin-induced diabetes, circulating EPCs were reduced to 39 +/- 5% of controls and associated with uncoupled eNOS in bone marrow. Our results identify uncoupling of eNOS in diabetic bone marrow, glucose-treated EPCs, and EPCs from diabetic patients resulting in eNOS-mediated O(2)(-) production. Subsequent reduction of EPC levels and impairment of EPC function likely contributes to the pathogenesis of vascular disease in diabetes.     

褪黑素抗氧化治疗糖尿病大鼠阴茎勃起功能障碍的研究

目的 观察褪黑素对糖尿病大鼠阴茎勃起功能的影响,探讨氧化应激在糖尿病性勃起功能障碍发病机制中的作用.方法 一次性腹腔注射STZ建立糖尿病大鼠模型,随机分为糖尿病组、褪黑素(MT)治疗组以及对照组.8周后通过电刺激各组大鼠勃起神经来检测海绵体内压,评价勃起功能;采用硫代芭比妥酸法检测阴茎海绵体组织中丙二醛(MDA)含量,黄嘌呤氧化酶法测超氧化物氧化酶(SOD)活性;免疫组化染色半定量分析各组大鼠阴茎海绵体中平滑肌及内皮的含量.结果 与正常对照组相比,阴茎海绵体组织中MDA含量显著增加(P＜0.01),SOD活性降低(P＜0.05),最大海绵体内压(ICP)亦显著降低(P＜0.05);与糖尿病组相比,MT组大鼠海绵体MDA含量明显降低(P＜0.05),其SOD活性和ICP显著升高(P＜0.05);且其海绵体平滑肌及海绵窦内皮细胞含量明显提高.结论 MT可通过改善组织中氧化应激水平,促进阴茎海绵体平滑肌和内皮组织修复,提高勃起功能;抗氧化治疗可能为糖尿病性勃起功能障碍的防治提供新的策略.     

缺血性糖尿病足部溃疡皮肤周边组织Vδ1T细胞IGF-1表达与MDA和SOD生成的研究

目的:探讨缺血性糖尿病足部溃疡皮肤Vδ1T细胞中胰岛素样生长因子-1(insulin-like growth factor,IGF-1)的表达及其临床意义及溃疡皮肤周边组织丙二醛(malonal dehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)生成的研究。方法:双酶标免疫荧光标记Vδ1T细胞及IG F-1蛋白的表达,以Olympus Fluo View FV10i激光共聚焦显微镜观察和摄像,并用IMAGE-PROPLUS 6.0分析IGF-1的表达。硫代巴比妥酸法测定MDA,黄嘌呤氧化酶法测定。实验分组:2008年10月～2010年年11月在湖南省郴州市第一人民医院住院和门诊的20例患者自愿捐献的标本,分为两组,每组10人,缺血性糖尿病足部溃疡组:男5例,女5例;正常足部皮肤对照组:正常对照组:男6例,女4例。结果:缺血性糖尿病足部溃疡组Vδ1T细胞IGF-1的表达较正常对照组减少,差异有统计学意义(P<0.001),溃疡周边皮肤组织MDA生成较正常对照组增加(P<0.001),而SOD较正常组减少(P<0.001)。结论:缺血性糖尿病足部溃疡皮肤Vδ1T细胞中IGF-1的表达减少,溃疡周边组织MDA生成增多,SOD生成减少,提示Vδ1T细胞中IGF-1的减少可能与缺血性糖尿病足部溃疡难以愈合有关,并且IGF-1减少与溃疡周边组织氧化应激有关。     

还原型谷胱甘肽对糖尿病大鼠阴茎海绵体氧化损伤的影响

目的探讨氧化损伤在糖尿病性ED发病机制中的作用,以及还原型谷胱甘肽（GSH）对氧化损伤的干预作用,为临床治疗提供新的手段。方法成年雄性SD大鼠50只,随机分为4组：A组为对照组（n=10）,B组为糖尿病无干预组（n=13）,C组为糖尿病胰岛素干预组（n=13）和D组为糖尿病胰岛素＋GSH干预组（n=14）。采用腹腔注射STZ法制作糖尿病模型并进行筛选。8周后取大鼠阴茎海绵体组织进行组织匀浆,采用黄嘌呤氧化法测定阴茎海绵体组织中超氧化物岐化酶（SOD）活性;用TBA法测定丙二醛（MDA）含量;RT—PCR法检测胞外型SOD基因（SODEX基因）的表达;Western-blot法检测CuZnSOD蛋白含量。结果A组大鼠海绵体组织基本无过氧化产物蓄积。与A组相比,B、C、D组的SOD活性显著下降（P〈0．05）,MDA含量均显著升高（P〈0．05）,B、C、D组SODEX基因的表达、CuZnSOD蛋白含量显著下降（P〈0．05）;与B组相比,C、D组的SOD活性显著升高（P〈0．05）,MDA含量显著下降（P〈0．05）,SODEx基因的表达及CuZnSOD蛋白含量显著升高（P〈0．05）。与C组相比,D组SODEX基因的表达及CuZnSOD蛋白含量显著升高（P〈0．05）。结论糖尿病性雄性大鼠阴茎海绵体组织自由基代谢明显障碍,脂质过氧化产物增加,同时氧自由基清除能力下降;而抗氧化剂GSH的应用能部分减弱氧化损伤。提示氧化损伤可能在糖尿病性ED的发生发展中起一定作用。本研究为糖尿病性ED的预防和治疗提供新的策略。     

Role of Irbesartan on cardiac endothelial - to - mesenchymal transition in diabetic rats

Objective To explore the effect of irbesartan on cardiac endothelial-mesenchymal transition (EndMT) in diabetic rats. Methods The model of diabetic rat was induced by intraperitoneal injection with streptozotocin (STZ, 35 mg/kg) in spontaneous hypertensive rats (SHR). Diabetic rats were divided into diabetic group and the Irbesartan treated group. The pathological changes were investigated by fluorescence microscope and electron microscope. The EndMT was studied in human aortic endothelial cells (HAEC) exposure to high glucose. The concentration of angiotensin II in the supernatant was detected by radioimmunoassay. Immunofluorescence staining was performed to detect the co - localization of CD31 and FSP1. Results The significant myocardial fibrosis was presented in the diabetic group. Endothelial protrusions were prominent feature in myocardial microvascular of diabetic rat compared with the control group rats. Double staining of HAEC showed co-localization of CD31 and FSP1, which was decreased by the treatment of Irbesartan (P＜0.05). When HAEC was exposed to high glucose, it showed some cells acquired spindle-shaped morphology and lost CD31 staining, and FSP1 and α - SMA protein expression levels were markedly upregulated, which attenuated by the treatment of Irbesartan. Conclusion Irbesartan might prevent diabetes from myocardial fibrosis via inhibition of EndMT in diabetic rats.     

Co‐Transplantation of Bone Marrow‐Derived Endothelial Progenitor Cells Improves Revascularization and Organization in Islet Grafts

Bone marrow‐derived early endothelial progenitor cells (BM‐EPCs) are a clinical tool for enhancing revascularization. However, the therapeutic efficacy of co‐transplantation of BM‐EPC with islets has not been investigated. In this study, marginal mass islets were co‐transplanted with or without BM‐EPCs under the kidney capsules of syngeneic streptozotocin‐induced diabetic mice. Using green fluorescent protein transgenic (GFP‐Tg) mice as BM‐EPC and islet donors or recipients, the role of EPCs in revascularization was assessed for graft morphology, vascular density and fate of EPCs by immunohistochemistry. Islet‐EPC co‐transplantation improved the outcome of islet transplantation as measured by glucose tolerance, serum insulin level and diabetes reversal rate, compared with transplantation of islets alone. Between groups, the morphology of islet grafts showed significant differences in size and composition of grafted endocrine tissues. Significantly more vessel density derived from donors and recipients was detected with islet‐EPC co‐transplantation. Abundant GFP‐Tg mice‐derived BM‐EPCs (GFP‐EPCs) were observed in or around islet grafts and incorporated into CD31‐positive capillaries. Remaining GFP‐EPCs expressed VEGF. In conclusion, co‐transplantation of islets with BM‐EPCs could improve the outcome of marginal mass islet transplantation by promoting revascularization and preserving islet morphology.