内皮素和一氧化氮及其在糖尿病肾病中的作用

糖尿病肾病是导致终末期肾衰的常见病因，早期以肾小球高滤过，小肥大，基膜增厚为特征最终导致肾小球硬化，在诸多介导糖尿病肾损伤的因素中，内皮素和一氧化氮被认为是重要的因子。内皮素通过其生长因子作用促进肾细胞增殖及胞外基质蛋白的合成，促进肾小球硬化，一氮化氮则通过扩张肾入球小动脉，在介导糖尿病肾病早期肾小球高滤过中起了重要作用，本文就这些方面的进展进行综述。     

内皮素和一氧化氮及其在糖尿病肾病中的作用

糖尿病肾病是导致终末期肾衰的常见病因 ,早期以肾小球高滤过 ,小球肥大 ,基膜增厚为特征最终导致肾小球硬化 ,在诸多介导糖尿病肾损伤的因素中 ,内皮素和一氧化氮被认为是重要的因子。内皮素通过其生长因子作用促进肾细胞增殖及胞外基质蛋白的合成 ,促进肾小球硬化。一氮化氮则通过扩张肾入球小动脉 ,在介导糖尿病肾病早期肾小球高滤过中起了重要作用 ,本文就这些方面的进展进行综述     

一氧化氮与糖尿病肾病

糖尿病肾产现是糖尿病致残，致死的主要病因之一，但上前尚未明确其发病机制。本文综述了一氧化氮对入，出血球小动脉及系膜细胞的作用；ＮＯ水平升高引起肾小球高灌注、高滤过，与ＤＮ密切相关。     

一氧化氮与糖尿病肾病

糖尿病内源性ＮＯ变化可能是一个双向过程且对ＤＮ的发生发展具有重要影响，ＮＯ的升高参与了肾小球高滤过的机制，而ＮＯ的下降则加剧了肾小球病理形态学改变。ＮＯ的动态变化机制与氧自由基、ＡＧＥ、ＡＲ及ＰＫＣ有关。     

一氧化氮与糖尿病肾病

糖尿病肾病(DN)是糖尿病致残、致死的主要病因之一,但目前尚未明确其发病机制。本文综述了一氧化氮(NO)对人、出球小动脉及系膜细胞的作用;NO水平升高引起肾小球高灌注、高滤过,与DN密切相关。     

一氧化氮与糖尿病肾病

糖尿病内源性ＮＯ变性可能是一个双向过程且对ＤＮ的发生发展具有重要影响，ＮＯ的升高参与了肾小球高滤过的机制，而ＮＯ的下降则加剧了肾小球病理形态学改变。ＮＯ的动态变化机制与氧自由基，ＡＧＥ，ＡＲ及ＰＫＣ的有关。     

内皮素在糖尿病肾病中的作用研究

糖尿病肾病时具有血浆、肾脏局部及尿液内皮素（ET）含量变化。ET通过干扰正常血糖代谢、影响体内其他细胞因子与生长因子以及改变肾脏血流动力、调节水盐代谢等机理参考糖尿病肾病的发生与进展。     

血红蛋白在鉴别糖尿病肾病与非糖尿病肾脏疾病中的作用

目的探讨血红蛋白在鉴别糖尿病肾病(DN)与非糖尿病肾脏疾病(NDRD)中的临床意义。 方法对2004年1月至2012年4月在解放军总医院肾脏病科行肾活检、病理诊断明确且病史资料完整的66例DN、78例NDRD患者进行回顾性分析,比较两组临床指标及合并症的差异,采用多因素Logistic回归分析方法明确DN的独立危险因素;比较DN及NDRD组不同CKD分期的血红蛋白水平差异,明确其贫血性质及贫血相关的独立危险因素。 结果DN组糖尿病罹病时间、肾病罹病时间、平均动脉压、血清肌酐、尿素氮、24 h尿蛋白定量水平及合并糖尿病视网膜病变、心血管病、贫血的比例均高于NDRD组,差异具有统计学意义(P<0.05);多因素分析显示糖尿病罹病时间长(OR＝1.012,95%CI 1.005～1.019)、合并糖尿病视网膜病变(OR＝ 4.265,95%CI 1.616～11.255)、血红蛋白水平低(OR＝ 0.952,95%CI 0.929～0.976)及合并心血管病(OR＝2.875,95%CI 1.089～7.593)是DN的独立危险因素;CKD1～3期DN组及NDRD组的血红蛋白水平均存在显著性差异(P<0.05),该差异在CKD 4～5期消失;DN及NDRD组共有60例诊断为贫血,均为正细胞正色素性贫血;多因素Logistic回归分析显示白蛋白降低(OR＝0.928,95%CI 0.879～0.980)、血清肌酐升高(OR＝1.011,95%CI 1.004～1.019)、病理诊断为DN(OR＝6.213,95%CI 2.690～14.347)是贫血的独立危险因素。 结论血红蛋白与DN显著独立相关,早期对血红蛋白水平的监测可能为临床鉴别DN与NDRD提供新线索。     

川芎嗪联合氨胍对糖尿病大鼠肾脏一氧化氮的影响

目的：探讨川芎嗪联合氨胍对糖尿病肾脏病变的保护作用及其机制。方法：用链脲佐菌素制作糖尿病大鼠模型，分为正常对照组、糖尿病模型组、川芎嗪治疗组、氨胍治疗组和川芎嗪联合氨胍治疗组，于第12周测定各组大鼠肾组织一氧化氮合酶的活性和一氧化氮含量。结果：川芎嗪联合氨胍治疗纪、川芎嗪治疗组、氨胍治疗组大鼠肾组织NOS活性显高于模型组(P＜0．01)，NO的含量增加(P＜0．01)。川芎嗪治疗组、氨胍治疗组NOS活性低于正常对照组(P＜0．01)，NO的合量降低(P＜0．01)；川芎嗪合氨基胍治疗组与正常组相比无差异。结论：川芎嗪联合氨基胍能够增强糖尿病大鼠肾组织NOS活性，增加NO的含量，从而对糖尿病性肾病起一定治疗作用。     

一氧化氮在门静脉高压症发病中的作用

目的　探讨一氧化氮在门静脉高压症发病中的作用。　方法　 75例门静脉高压症患者 ,术中胃网膜静脉插管测定门静脉压力 ,检测外周动静脉和门静脉血中内毒素和NO2 -/NO3-的含量。　结果　 ( 1)门静脉高压症患者的血中内毒素和NO2 -/NO3-的水平 [( 0 2 4 9± 0 112 )Eu/ml和( 5 5 9± 2 6 2 ) μmol/L]均显著高于对照组 ,且门静脉血中水平最高。 ( 2 )门静脉高压症患者门静脉压力[( 3 5 5± 4 4 )cmH2 O]与门静脉血NO2 -/NO3-的水平呈显著正相关 (n =2 5 ,r =0 5 5 ,P <0 0 1) ,二者在术后的变化量也呈正相关 (r =0 5 7,P <0 0 5 )。 ( 3 )门静脉高压症患者白蛋白水平与NO2 -/NO3-呈负相关 (n =75 ,r=- 0 3 5 ,P <0 0 1) ,且有腹水组的NO2 -/NO3-水平 [( 72 4± 2 0 3 ) μmol/L]较无腹水组 [( 5 0 3± 2 1 0 ) μmol/L]为高。　 结论　门静脉高压症患者血中内毒素和NO的水平升高 ,后者可能参与了门静脉压力的异常升高且与肝功能损害有关。     

Role of nitric oxide in diabetic nephropathy

Diabetic nephropathy is the leading cause of end-stage renal disease in the Western hemisphere. Endothelial dysfunction is the central pathophysiologic denominator for all cardiovascular complications of diabetes including nephropathy. Abnormalities of nitric oxide (NO) production modulate renal structure and function in diabetes but, despite the vast literature, major gaps exist in our understanding in this field because the published studies mostly are confusing and contradictory. In this review, we attempt to review the existing literature, discuss the controversies, and reach some general conclusions as to the role of NO production in the diabetic kidney. The complex metabolic milieu in diabetes triggers several pathophysiologic mechanisms that simultaneously stimulate and suppress NO production. The net effect on renal NO production depends on the mechanisms that prevail in a given stage of the disease. Based on the current evidence, it is reasonable to conclude that early nephropathy in diabetes is associated with increased intrarenal NO production mediated primarily by constitutively released NO (endothelial nitric oxide synthase [eNOS] and neuronal nitric oxide synthase [nNOS]). The enhanced NO production may contribute to hyperfiltration and microalbuminuria that characterizes early diabetic nephropathy. On the other hand, a majority of the studies indicate that advanced nephropathy leading to severe proteinuria, declining renal function, and hypertension is associated with a state of progressive NO deficiency. Several factors including hyperglycemia, advanced glycosylation end products, increased oxidant stress, as well as activation of protein kinase C and transforming growth factor (TGF)-beta contribute to decreased NO production and/or availability. These effects are mediated through multiple mechanisms such as glucose quenching, and inhibition and/or posttranslational modification of NOS activity of both endothelial and inducible isoforms. Finally, genetic polymorphisms of the NOS enzyme also may play a role in the NO abnormalities that contribute to the development and progression of diabetic nephropathy.     

Chronic diabetic nephropathy: role of inducible nitric oxide synthase

Nitric oxide (NO) is a multifunctional mediator that has been implicated in the short-term hemodynamic alterations that occur in acute streptozocin (STZ)-induced diabetes. We investigated the role of NO produced by inducible nitric oxide synthase (iNOS) in chronic STZ diabetic nephropathy. Diabetes was induced in C57BL/6 and iNOS knockout (KO) mice with two intraperitoneal injections of STZ, 100 mg/kg. Animals were maintained without insulin treatment for 40 weeks. There were no significant differences between the strains in blood urea nitrogen (BUN), serum creatinine or glucose concentration, or urinary protein excretion during the entire observation period. Urinary nitrite + nitrate excretion was significantly lower in iNOS KO mice compared to control animals at all time points; in C57 mice, urinary nitrite declined progressively with more prolonged duration of diabetes. Renal hypertrophy (kidney weight/body weight) was noted in both strains of mice. However, histopathological assessment of renal tissue specimens at 16 and 40 weeks demonstrated increased mesangial hypercellularity and expansion as well as more prominent tubulointerstitial fibrosis in iNOS KO versus C57 mice. These changes were accompanied by increased interstitial deposition of type I collagen at 16 and 40 weeks in iNOS KO mice. Glomerular basement membrane staining for type IV collagen was also increased at 40 weeks in diabetic iNOS KO mice. While iNOS protein was undetectable in any of the kidney specimens obtained from either strain, eNOS was present throughout the course of chronic STZ diabetes. Moreover, eNOS expression was significantly increased by approximately 40% at 16 and 40 weeks of observation in iNOS KO versus C57 mice. There was no difference in renal cortical malondialdehyde content between the strains early or late in the disease course. In time control animals, there was no evidence of renal histopathological damage in iNOS KO or C57 mice after 40 weeks. We conclude that iNOS-derived NO modulates glomerulosclerosis and tubulointerstitial fibrosis in chronic STZ nephropathy. This action is probably a result of the direct actions of NO on the synthesis and degradation of extracellular matrix proteins. Received: 28 February 2001 / Revised: 10 August 2001 / Accepted: 13 August 2001     

Associations between endothelial nitric oxide synthase polymorphisms and risk of diabetic nephropathy: an updated meta-analysis

Abstract

Background: Genetic polymorphism of endothelial nitric oxide synthase (eNOS) has been implicated in the risk of diabetic nephropathy (DN), but the published findings were inconsistent. We performed a comprehensive meta-analysis to derive a more precise estimation of the association between genetic polymorphisms of eNOS and the risk of DN. Methods: Six online database were researched on the associations between polymorphisms of eNOS (T786C, G894T, 4b/4a) and DN risk. PRISMA statement and Hardy–Weinberg equilibrium assessment were used in this report. Odds ratio and 95% confidence interval were estimated based on the following genetic contrast/models: allelic model, dominant model, recessive model and co-dominant model. The publication bias and sensitivity analysis were also performed to guarantee the statistical power. Results: A total of 49 case–control studies with 11,990/9754/5131 participants for DN/DM/HC group were eligible for meta-analysis (7/25/31 studies for T786C/G984T/4b/a). For the eNOS-T786C, C allele showed a weak association between C allele and DN risk in DN/T2DM group. For eNOS-G894T, there was an association between T allele and DN risk in the global, Asian and African population in DN/T2DM group. For the eNOS-4b/4a, 4a allele was found contributing significantly to increased DN risk in the global population. Conclusion: Our comprehensive meta-analysis suggests that three polymorphisms of eNOS may be the increased risk factors of DN development, especially in Asian population and T2DM group.     

内皮型一氧化氮合酶在大鼠糖尿病膀胱病变中的作用

目的 探讨大鼠糖尿病膀胱病变组织中内皮型一氧化氮合酶(eNOS)的表达及意义.方法 雄性SD大鼠36只.随机分为正常组、多尿组和糖尿病组,每组12只.采用尿动力学方法对大鼠膀胱容量、单次排尿量、膀胱内最大压力、残尿壁和排尿率等指标进行评价,应用RT-PCR和蛋白质印迹方法检测大鼠膀胱组织中eNOS mRNA和蛋白质的表达水平,统计学分析其与膀胱功能的相关性.结果 6周时正常组大鼠最大膀胱容量、单次排尿量、残余尿量分别为(0.75±0.04)、(0.70±0.02)和(0.06±0.00)ml,多尿组分别为(1.62±0.15)、(1.52±0.30)和(0.11±0.01)ml,糖尿病组分别为(2.29±0.16)、(2.06±0.25)和(0.23±0.01)ml,与正常组和多尿组相比,差异具有统计学意义(P＜0.01);12周时正常组上述指标分别为(0.86±0.06)、(0.80±0.04)和(0.07±0.00)ml,多尿组分别为(1.93±0.10)、(1.77±0.11)和(0.13±0.02)ml,糖尿病组分别为(2.65±0.26)、(2.09±0.21)和(0.56±0.06)ml,各组间差异具有统计学意义(P＜0.01).糖尿病组与6周时相比,各指标增加,其中残余尿量差异具有统计学意义(P＜0.01).6周时3组排尿率分别为(93.3±2.1)%、(93.2±2.7)%和(90.0±2.5)%,组间差异无统计学意义(P＞0.05);12周时分别为(93.1±2.2)%、(93.8±3.2)%和(78.1±2.6)%,糖尿病组小于其他2组(P＜0.05).各组膀胱内压力差异无统计学意义.糖尿病组膀胱压力曲线出现压力小波动,曲线不规则.6周时各组膀胱组织中eNOSmRNA表达相对灰度值分别为0.81±0.12、0.90±0.12和1.98±0.16,糖尿病组高于其他2组,差异具有统计学意义(P＜0.01);12周时为0.87±0.17、1.05±0.11和2.89±0.23,糖尿病组与6周时相比增加(P＜0.01).6周时各组eNOS蛋白表达相对灰度值分别为0.98±0.12、0.93±0.14和2.28±0.16,糖尿病组高于其他2组(P＜0.01);12周时为1.03±0.13、1.14±0.11和3.12±0.20,糖尿病组与6周时相比增加(P＜0.01).糖尿病组大鼠膀胱eNOS蛋白表达强度与排尿率旱负相关(r=-0.669,P＜0.01).结论 膀胱组织中eNOS mRNA和蛋白表达上调可能参与了糖尿病膀胱病变的发生发展.     

糖尿病患者血清一氧化氮水平对骨代谢影响

目的探讨糖尿病患者血清一氧化氮水平对成骨细胞、Ⅰ型骨胶原代谢的作用。方法 对确诊1型和2型糖尿病的378例患者测定血清一氧化氮(NO)、一氧化氮合酶(NOS)(硝酸还原酶法)、骨钙素《BGP放免法)、尿脱氧吡啶啉(DPD化学发光法)、尿肌酐(Cr酶法),双能X线测定腰椎正位、前臂、髋部骨密度(BMD),并与同期体检的342例健康人群进行比较。结果 糖尿病患者血清NO、NOS显著高于对照组(P<0.001;P<0.05),BGP显著低于对照组(P<0.001),DPD／Cr则显著高于对照组(P<0.001)。糖尿病组和对照组NO与BGP、DPD及各部位BMD均无显著线性相关(P>0.05)。结论糖尿病患者成骨细胞代谢显著低于健康人而Ⅰ型骨胶原分解代谢则显著高于健康人,血清一氧化氮水平与成骨细胞和Ⅰ型骨胶原代谢无显著相关性。     

Diabetic endothelial nitric oxide synthase knockout mice develop advanced diabetic nephropathy

The pathogenesis of diabetic nephropathy remains poorly defined, and animal models that represent the human disease have been lacking. It was demonstrated recently that the severe endothelial dysfunction that accompanies a diabetic state may cause an uncoupling of the vascular endothelial growth factor (VEGF)-endothelial nitric oxide (eNO) axis, resulting in increased levels of VEGF and excessive endothelial cell proliferation. It was hypothesized further that VEGF-NO uncoupling could be a major contributory mechanism that leads to diabetic vasculopathy. For testing of this hypothesis, diabetes was induced in eNO synthase knockout mice (eNOS KO) and C57BL6 controls. Diabetic eNOS KO mice developed hypertension, albuminuria, and renal insufficiency with arteriolar hyalinosis, mesangial matrix expansion, mesangiolysis with microaneurysms, and Kimmelstiel-Wilson nodules. Glomerular and peritubular capillaries were increased with endothelial proliferation and VEGF expression. Diabetic eNOS KO mice showed increased mortality at 5 mo. All of the functional and histologic changes were improved with insulin therapy. Inhibition of eNO predisposes mice to classic diabetic nephropathy. The mechanism likely is due to VEGF-NO uncoupling with excessive endothelial cell proliferation coupled with altered autoregulation consequent to the development of preglomerular arteriolar disease. Endothelial dysfunction in human diabetes is common, secondary to effects of glucose, advanced glycation end products, C-reactive protein, uric acid, and oxidants. It was postulated that endothelial dysfunction should predict nephropathy and that correction of the dysfunction may prevent these important complications.     

Endothelial nitric oxide synthase deficiency produces accelerated nephropathy in diabetic mice

Functionally significant polymorphisms in endothelial nitric oxide synthase (eNOS) and reduced vascular eNOS activity have been associated with increased human diabetic nephropathy (DN), but the pathogenic role of eNOS deficiency in the development of DN has not yet been confirmed. This study characterizes the severity of DN in eNOS(-/-) mice that were backcrossed to C57BLKS/J db/db mice. Although the severity of hyperglycemia was similar to C57BLKS/J db/db mice, by 26 wk, eNOS(-/-) C57BLKS/J db/db mice exhibited dramatic albuminuria, arteriolar hyalinosis, increased glomerular basement membrane thickness, mesangial expansion, mesangiolysis, and focal segmental and early nodular glomerulosclerosis. Even more remarkable, eNOS(-/-) C57BLKS db/db exhibited decreases in GFR to levels <50% of that in eNOS(+/+) C57BLKS db/db, as confirmed by increased serum creatinine. In summary, eNOS(-/-) db/db mice provide the most robust model of type II DN that has been described to date and support a role for deficient eNOS-derived NO production in the pathogenesis of DN.     

糖尿病肾病的蛋白质组学研究进展

糖尿病肾病(DN)是糖尿病最常见的微血管并发症,也是导致终末期肾病(ESRD)的重要原因。微量白蛋白尿作为预测DN进展的指标并不可靠,寻找DN早期生物学标志物以及对DN早期肾脏病变的机制研究为目前研究的重点。蛋白质组学技术近年来飞速发展,对DN患者的尿液、血液以及肾组织的蛋白质组学研究可能会为我们早期诊治DN提供新的希望。本文对蛋白质组学主要技术及DN蛋白质组学研究的近期研究进展进行综述。     

洛沙坦对糖尿病大鼠肾组织一氧化氮水平的影响

目的 研究洛沙坦对糖尿病大鼠肾组织一氧化氮（NO）水平的影响。方法 雄性Wistar大鼠分为3组,A组（11只）为正常对照组,B组（11只）为糖尿病未干预组,C组（9只）为糖尿病大鼠洛沙坦干预组。以链脲菌素制备糖尿病大鼠模型,大鼠饲养18周后取出肾脏检测诱导型NO合成酶（iNOS）mRNA的表达,电镜检测大鼠肾小球基底膜厚度及系膜基质密度（系膜基质面积/系膜面积）。收集24h尿测定尿白蛋白排泄（UAE）及肌酐,并心脏内取血检测血肌酐。mRNA表达采用RT-PCR,以β-actin作为内对照。UAE测定采用大鼠白蛋白特异的酶免疫分析试剂盒。结果 肾组织iNOSmRNA表达在B组大鼠（0.30±0.12）显著高于A组（0.12±0.04,P<0.01）,C组（0.25±0.14）与B组比较差异无显著性意义（P>0.05）。肾组织NO水平在B组大鼠[（0.56±0.20）μmol/mg肾组织]显著低于A组[（1.05±0.25）μmol/mg肾组织]和C组[（1.13±0.62）μmol/mg肾组织,P均<0.01]。UAE在B组大鼠[（2.18±1.98）mg/d]显著高于A组[（0.41±0.47）mg/d]和C组[（0.65±0.89）mg/d,P均<0.05]。肌酥清除率在B组大鼠[（19.75±9.60）ml/d]显著低于A组[（59.63±22.75）ml/d]和C组[（40.88±25.57）ml/d,P均<0.05]。基底膜厚度在B组大鼠[（531.6±107.6）nm]显著高于A组[（312.4±25     

氯沙坦钾对糖尿病肾病大鼠血浆一氧化氮和肝细胞生长因子水平的影响

本研究旨在观察氯沙坦钾对糖尿病肾病(DN)大鼠血浆一氧化氮(NO)和肝细胞生长因子(HGF)水平的影响, 从而探讨氯沙坦钾对DN的肾脏保护作用机制.