白细胞介素22抗体抑制Snail1高表达对糖尿病肾病的影响

目的:探讨白细胞介素22(IL-22)在糖尿病肾病(DN)发生发展中的作用及机制。方法:将C57 BL/6小鼠随机分成正常对照(NC)组、DN组、重组IL-22(r IL-22)干预组(DN+r IL-22组)和IL-22中和抗体(anti-IL-22)干预组(DN+anti-IL-22组)。糖尿病造模成功8周后,DN+rIL-22组和DN+anti-IL-22组分别给予rIL-22(200μg/kg)和anti-IL-22(200μg/kg)腹腔注射,NC组和DN组分别给予等量的0.1%牛血清白蛋白腹腔注射,每周2次,连续4周。干预结束后,检测各组小鼠血糖、肾功能、24 h尿微量白蛋白(m-Alb)和24 h尿肌酐(UCr)水平,光镜下观察肾脏组织的病理结构改变,qPCR法检测肾脏组织Snail1的mRNA表达,Western blot法检测肾脏组织纤连蛋白(FN)和E-钙黏素(E-cadherin)的表达水平。结果:干预结束后,与NC组小鼠比较,各模型组24 hm-Alb/UCr比值均显著升高(P0.05);相较于DN组,DN+rIL-22组24 hm-Alb和24 hUCr均显著升高(P0.05);而DN+anti-IL-22组24 hm-Alb、24 h UCr和24 hm-Alb/UCr比值较DN组和DN+rIL-22组均得到改善(P0.05)。光镜下可见糖尿病小鼠肾小管上皮细胞空泡变性、蛋白管型形成和肾小球系膜扩张,DN+rIL-22组上述病变更广泛,而DN+anti-IL-22组病变程度得以改善。qPCR发现糖尿病小鼠Snail1的mRNA表达显著升高(P0.05),anti-IL-22阻断4周后Snail1的mRNA显著下降(P0.05)。Western blot发现DN+rIL-22组细胞外基质蛋白FN较NC组和DN组显著升高(P0.05),且肾小管上皮-间充质转化标志物E-cadherin显著下降(P0.05)。结论:IL-22中和抗体可能通过抑制肾小管上皮细胞Snail1信号分子高表达,减轻DN小鼠微量白蛋白尿,延缓DN进程。     

贝那普利和缬沙坦联合治疗糖尿病肾病的疗效观察

目的：观察贝那普利联合缬沙坦治疗糖尿病肾病临床疗效。方法：68例符合入选条件的患者被随机分为三组,分别进行贝那普利,缬沙坦以及贝那普利和缬沙坦联合用药治疗,血压控制在130／80mmHg以下,疗程为12周,治疗前后测定24小时尿蛋白定量,同时测定肾功能、血糖、血钾。结果：治疗12周后,各组治疗前后血压均显著下降（P〈0．05）,24小时尿蛋白定量治疗后较治疗前均有显著下降,且联合治疗组尿蛋白下降程度明显优于贝那普利组与缬沙坦组,差异有显著性意义（P＜0．01）。结论：贝那普利和缬沙坦联合用药降低糖尿病肾病患者尿蛋白疗效优于单一用药,且副作用少。     

Animal model of diabetic nephropathy

Injection of subtotally nephrectomized rats with streptozotocin produced metabolic abnormalities resembling diabetic nephropathy in humans. These abnormalities were hyperglycemia, hypoinsulinemia, azotemia, hypertriglyceridemia and hypercholesterolemia, accompanied with an increase in glycosylated protein. Extraordinary changes in the urinary excretion of glucose and protein were also observed in rats that received streptozotocin treatment after subtotal nephrectomy. In addition, the level of creatinine clearance was significantly decreased. The pathological findings in the kidney of these rats revealed lesions of the glomerular capillary loops, mesangial area and Bowman's capsule. Coagulation was also found in the glomerular capillaries. Our results suggest that this rat model would be useful for studies of diabetic nephropathy.     

随机尿微量白蛋白检测对筛检早期糖尿病肾病的价值

评价随机尿白蛋白浓度检测对筛选糖尿病微量白蛋白尿的效能.198例2型糖尿病患者纳入研究,完成其24h尿样收集及白天随机尿收集.白蛋白测定采用免疫比浊法.根据24h尿白蛋白排泄率(24h UAER)分为正常白蛋白尿组(24h UAER＜20μg/min,102例)及微量白蛋白尿组(24h UAER 20-200μg/min,96例)两组.并绘制随机尿白蛋白浓度(UAC)受试者工作曲线(ROC).结果是:UAC与UAER相关(r=0.92,P＜0.001),UAC-ROC 100%敏感度截断点UAC值为15mg/L(特异度为74%),敏感度与特异度最佳截断点UAC值为30mg/L(敏感度90%,特异度89%).随机UAC对于筛选微量白蛋白尿准确性较高,且较UAER简便、经济,适用于筛检早期糖尿病肾病.     

ACE2内源性激动剂DIZE对糖尿病肾病大鼠的保护作用

目的:观察血管紧张素转换酶2(ACE2)内源性激动剂乙酰甘氨酸重氮氨苯脒(DIZE)对糖尿病肾病(DN)大鼠的保护作用。方法:30只Wistar大鼠随机分为正常对照组(NC组)、DN组和DIZE处理组(DIZE组)。DN组与DIZE组一次性腹腔注射链脲佐菌素(65 mg/kg)建立糖尿病模型,12周后糖尿病肾病大鼠模型建立后给予DIZE 15 mg·kg~(-1)·d~(-1)或等量生理盐水皮下注射4周处理。16周末称量体重和肾重,计算肾质量体质量比(KW/BW),收集血、尿标本,检测血糖(GLU)、24 h尿蛋白(24UP)及血清肌酐(SCr)等指标。通过PAS染色观察各组肾脏病理变化;ELISA法检测大鼠AngⅡ、Ang-(1-7)、TGF-β1及VCAM-1水平的变化;通过免疫组化观察collagenⅠ和FN蛋白表达的变化;利用实时荧光定量PCR(RT-qPCR)技术检测大鼠肾组织collagenⅠ和FN mRNA含量的变化;Western blot观察各组大鼠ACE2蛋白表达的变化。结果:DIZE显著提高了糖尿病大鼠ACE2的表达(P0.05),降低了糖尿病大鼠血浆AngⅡ含量(P0.05),提高了Ang-(1-7)的水平(P0.05)。与NC组大鼠相比,DN组与DIZE组大鼠的24UP、SCr和KW/BW明显升高(P0.05),collagenⅠ和FN mRNA水平及蛋白表达量增加,肾脏组织TGF-β1及VCAM-1明显上升(P0.05)。DIZE组与DN组大鼠相比,24UP和SCr水平降低(P0.05),GLU和KW/BW无明显差异,collagenⅠ和FN mRNA含量及蛋白表达量减少,肾脏组织TGF-β1及VCAM-1水平降低(P0.05)。结论:ACE2内源性激动剂DIZE显著提高了ACE2的活性,增加了Ang-(1-7)的含量,从而降低了肾脏纤维化及炎症水平,并对糖尿病肾病大鼠起到保护性作用。     

炎症与糖尿病肾病

近年来,炎症反应在糖尿病及其并发症发病机制中的作用引起了人们的广泛关注,研究报道炎症因子及促炎症因子与糖尿病肾病的发生和发展密切相关,并认为糖尿病肾病是一种炎症性疾病,炎症因子的大量产生参与并促进糖尿病肾病的进展。本文重点就炎症因子与糖尿病肾病的关系以及抗炎治疗对糖尿病肾的保护作用进行综述。     

血、尿IL-6变化在糖尿病肾病中的意义

目的 探讨血、尿IL-6变化在糖尿病肾病诊断治疗中的意义.方法 将实验对象分为观察组和对照组,观察组52人,对照组40人.根据尿白蛋白排泄率(UAER)将观察组又分为两个小组:分别为微量蛋白尿组DN1(24例)和临床蛋白尿组DN2(28例).对照组选取40名健康成年人.观察各组血、尿IL-6水平.结果 ①观察组病人血、...     

CXCL16缺失缓解STZ诱导的糖尿病小鼠的肾脏病变

 目的: 探索CXCL16基因缺失对链脲佐菌素(STZ)引发的糖尿病小鼠肾脏病变的影响。方法: 选取8周龄雄性C57BL/6J CXCL16基因缺失(C16 KO)小鼠,以及相同年龄及背景的野生型(WT)小鼠,采用STZ诱导的方式,建立糖尿病小鼠模型,观察CXCL16基因缺失对糖尿病肾病发生发展的影响。结果: 在糖尿病病变方面,与STZ处理的WT小鼠相比,STZ诱导C16 KO糖尿病小鼠的空腹血糖显著降低,并且其葡萄糖耐受能力得到显著改善。在糖尿病引发的肾脏病变方面,STZ处理后,C16 KO糖尿病小鼠的尿蛋白量显著低于WT糖尿病小鼠,此外,C16 KO糖尿病小鼠的肾小球损伤也明显低于WT糖尿病小鼠。与此同时,与WT糖尿病小鼠相比,C16 KO糖尿病小鼠肾脏组织活性氧簇(ROS)、丙二醛(MDA)、氧化低密度脂蛋白(ox-LDL)水平及凝集素样氧化低密度脂蛋白受体 1(Lox-1)的mRNA表达水平均显著下调。此外,在STZ处理后,C16 KO糖尿病小鼠肾组织中的巨噬细胞移动抑制因子(MIF)、炎症因子TNF-α 和IL-6以及黏附因子ICAM-1和CXCL1的mRNA表达水平均显著低于WT糖尿病小鼠。结论: CXCL16基因缺失能显著抑制STZ诱导的糖尿病小鼠肾脏病变。     

小檗碱对糖尿病肾病大鼠C反应蛋白的影响

目的:观察小檗碱对糖尿病肾病大鼠C反应蛋白的影响。方法:将48只雄性SD大鼠随机分为空白对照组、模型组、小檗碱75、150及300mg.kg-1组、罗格列酮组六组,每组8只。除正常对照组外,其余各组大鼠腹腔注射链脲佐菌素(STZ,55mg.kg-1),成功造模后小檗碱各组及罗格列酮组分别灌胃给予75、150、300mg.kg-1小檗碱以及4mg.kg-1罗格列酮,连续6w后检测大鼠血糖、体重,HE染色检测肾脏病理改变,ELISA法检测C反应蛋白活性。结果:与正常对照组相比,糖尿病组血糖与C反应蛋白活性明显升高(P<0.05),与模型组比较,小檗碱各组血糖与C反应蛋白活性明显降低(P<0.05),有统计学意义。结论:小檗碱可降低糖尿病大鼠血糖和C反应蛋白,减轻糖尿病肾病。     

Alterations of glomerular matrix proteins in the pathogenesis of diabetic nephropathy

Summary Diabetic late complications are characterized by morphological and biochemical alterations of the extracellular matrix. In particular, longstanding diabetes causes quantitative and qualitative changes in basement membrane structure of retinal and renal capilleries. Immunohistochemical investigations of diabetic kidneys with diffuse glomerulosclerosis show increased collagen type IV deposition in the mesangial matrix and decreased heparan sulfate proteoglycan content in the mesangial matrix and glomerular basement membrane as well. In nodular glomerulosclerosis normal basement membrane components are decreased or absent while the occurrence of collagen type III in this stage has been interpreted as an irreversible alteration of the glomerular structure. These changes seem to be the underlying cause for the alterations in renal functions like persistent albuminuria and proteinuria. Increased intra- and extracellular levels of glucose and its derivatives are thought to be responsible for diabetic tissue dysfunction although there are reports on possible genetic defects causing increased susceptibility to develop diabetic nephropathy. Recent results, however, focuse on the role of glucose-induced cytokine secretion as mediator for altered metabolism of glomerular matrix proteins. In vitro studies with cultured kidney cells have shown that the glucose-induced dysregulation of the basement membrane synthesis may be mediated by a glucose dependent activation of protein kinase C. Alternatively or synergistically, the formation of AGE products formed after prolonged exposure of matrix proteins to elevated glucose may also lead to cytokine secretion subsequently inducing synthesis of extracellular matrix proteins. Studies in experimental animals confirm the diabetes induced dysregulation of the synthesis of extracellular matrix components on the molecular level.Abbreviations HSPG heparan sulfate proteoglycan - GBM glomerular basement membrane - ECM extracellular matrix - AGE advanced glucosylation end products - TNF tumor necrosis factor - bFGF basic fibroblast growth factor     

微小RNA在糖尿病肾病发病机制及治疗中的研究进展

微小RNA（microRNAs,miRNA）可在转录后水平调控基因表达。最新研究表明miRNA与糖尿病肾病肾纤维化、足细胞损伤、系膜增生、微血管病变等密切相关,在糖尿病肾病的发病中可能起着必不可少的作用。深入研究miRNA可能为糖尿病肾病的诊断和治疗带来全新的方法。     

Human atrial natriuretic peptide in patients with type 1 diabetes mellitus: is it related to the development of diabetic nephropathy?

Summary There is controversy as to whether increased plasma levels of human atrial natriuretic peptide (hANP) in patients with type 1 diabetes mellitus may contribute to the development of diabetic nephropathy. Therefore, we decided to conduct two studies to examine the relationship of hANP levels to urinary albumin excretion and blood pressure. In a cross-sectional study, 83 randomly selected type 1 diabetic patients were investigated. 19 of the patients had increased urinary albumin excretion. 45 healthy volunteers served as controls. In a longitudinal study, 19 type 1 diabetic patients were examined for one year at monthly intervals. An increased risk of eventually developing diabetic nephropathy was identified in 7 out of these patients by repeatedly revealing increased urinary albumin excretion. On the average, hANP levels were increased in type 1 diabetic patients in comparison to controls (P < 0.001). In both studies, hANP levels were positively related (P < 0.05) to mean arterial blood pressure. There was no correlation between hANP levels and metabolic control. hANP levels lay within normal range irrespective of normal or elevated urinary albumin excretion provided that mean arterial blood pressure was normal. In the longitudinal study, increased urinary albumin and alpha-1-microglobulin excretion preceded the increase in both hANP levels and mean arterial blood pressure. Although hANP levels were evidently not related to the disease mechanisms of early diabetic nephropathy, it is tempting to speculate that hANP may contribute to the vicious circle connecting diabetic kidney disease to hypertension once that its levels are increased by elevated blood pressure.Abbreviation hANP human atrial natriuretic peptide     

糖尿病肾病防治新观点--美国第84届内分泌协会年会专题小结

糖尿病肾病是导致糖尿病死亡及终末期肾病透析的主要原因之一。在美国第84届内分泌协会年会上报告了一些糖尿病肾病治疗新观点。内容包括：HOPE研究结果；血管紧张素转换酶抑制剂(ACEI)是糖尿病患者心血管甚至是肾功能不全的保护药；ACEI可能能够预防糖尿病；血管紧张素1(AT1)受体阻滞剂(ARB)肾保护作用的证据；糖尿病中血压控制的重要性；ARB改善这些患者充血性心力衰竭及使用ACEI和ARB的临床建议等。本文对此作一综述。     

白藜芦醇对糖尿病肾病大鼠醛糖还原酶的影响

目的:观察白藜芦醇对糖尿病肾病大鼠醛糖还原酶的影响。方法:将48只雄性SD大鼠随机分为空白对照组、模型组、白藜芦醇低、中、高剂量组和依帕司他组(n=8)。除正常对照组外,其余各组大鼠腹腔注射STZ(55 mg.kg-1),成功造模后白藜芦醇组分别给予5、154、5 mg.kg-1白藜芦醇,依帕司他组给予10 mg.kg-1依帕司他,连续灌胃6 w后,检测大鼠血糖、体重,HE染色检测肾脏病理改变,ELISA法检测醛糖还原酶(AR)活性。结果:与正常对照组相比,糖尿病组血糖与AR活性明显升高(P<0.05),与模型组比较,白藜芦醇各组血糖与AR活性明显降低(P<0.05)。结论:白藜芦醇可降低糖尿病大鼠血糖,其降糖效应可能与抑制AR活性有关。     

左卡尼汀对糖尿病肾病大鼠的肾脏保护机制研究

目的:探讨左卡尼汀(L-carnitine,LC)对链脲佐菌素(streptozotocin,STZ)诱导的糖尿病肾病(diabetic nephropathy,DN)大鼠肾脏保护的分子机制。方法:对雄性Sprague-Dawley大鼠腹腔注射STZ(65 mg/kg)建立糖尿病模型,造模成功大鼠给予LC(50 mg·kg~(-1)·d~(-1)或200 mg·kg~(-1)·d~(-1))12周治疗,检测各组大鼠的肾功能、24 h尿蛋白排泄率和肾小球硬化程度;免疫组化和Western blot法分别检测炎性因子、致纤因子、核因子κB和细胞凋亡调控基因的表达。结果:LC治疗可明显减轻糖尿病大鼠肾小球硬化,保持足细胞数量,这些改变伴随着尿蛋白排泄率的减少和肾功能的改善。在分子水平上,LC可下调炎性介质和致纤因子的表达,调节细胞凋亡调控基因的表达,此作用可能与干预核因子κB信号通路有关。结论:LC对STZ诱导的DN大鼠具有抗炎、抗肾小球硬化的肾脏保护作用。     

Association of fibronectin Msp iv polymorphism and diabetic nephropathy susceptibility in Chinese Han population

Aim: Our study was aimed to study the distributional characteristics of fibronectin (Fn) Msp iv polymorphism in Chinese Han Population and investigate its association with susceptibility and clinicopathologic features of diabetic nephropathy (DN). Methods: Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were applied to testify Fn Msp iv genotypes among 108 patients with DN and 86 healthy individuals. Odds ratio (OR) with 95% confidence interval (CI) were used to evaluate the association of Fn Msp iv polymorphism and onset risk and clinicopathologic stages of DN. Results: The comparison of genotype and allele distribution in normal, micro and massive proteinuria groups showed that genotype and allele distribution in massive proteinuria group showed great differences, compared with those of control group (P = 0.006, P = 0.004). Further analysis on the association of Fn Msp iv polymorphism and occurrence of abnormal proteinuria suggested that DD genotype and D allele appeared to be a risk factor for abnormal proteinuria (OR = 3.553, 95% CI = 1.278-9.875; OR = 2.442, 95% CI = 1.378-4.327). Then, we analyzed the effects of Fn Msp iv polymorphism on the clinicopathologic stages of DN, the result showed that DD genotype showed great effect on the occurrence of early-onset DN (OR = 7.500, 95% CI = 1.691-33.272). For the DN patients with D allele, the risk for early-onset DN was increased 3.445 folds (OR = 4.445, 95% CI = 1.869-33.10.574). Conclusion: Fn Msp iv polymorphism appeared to be associated with DN susceptibility.     

Osteoinductive factor is a novel biomarker for the diagnosis of early diabetic nephropathy

Background: Microalbuminuria is the earliest clinical sign of diabetic nephropathy (DN). However, earlier markers as a diagnostic tool for DN was required for the invalid of microalbuminuria in some cases. Osteoinductive factor (OIF) was known to be an essential component of the normal vascular matrix. We aimed to research the relationship between DN and OIF, and discussed the availability of the serological markers for earlier stage of DN. Method: One hundred twenty Chinese subjects, who included patients with type 2 diabetes mellitus (T2DM), DN with microalbuminuria, and DN with macroalbuminuria, as well as healthy controls, were enrolled in this study. Serum OIF levels were examined by ELISA and other clinical biochemical parameters were tested based on standard methods. Results: Our results indicated that, serum OIF levels were significantly increased in DN subjects compared with healthy and T2DM subjects (P < 0.05 respectively). However, no significant changes in serum OIF levels were found between T2DM and healthy subjects. Furthermore, serum OIF had negative correlation with estimated glomerular filtration rate (eGFR) and positive correlation with blood urea nitrogen(BUN) and creatinine. ROC curve analysis showed that serum OIF level was a good sensitive and specificity marker for microalbuminuria and early renal damage with sensitivity of 86.7% and specificity of 95%, as well as for macroalbuminuria and damage progress with sensitivity of 90% and specificity of 95%. Conclusion: OIF may be an indicator of the earlier-stage DN in subjects with T2DM. Understanding the exact mechanism of up-regulated OIF in subjects with DN requires further study.     

Effects of captopril treatment versus placebo on renal function in type 2 diabetic patients with microalbuminuria: a long-term study

We evaluated the renal effect of long-term antihypertensive treatment (12 months) with the angiotensin-converting enzyme inhibitor captopril compared to placebo in 15 type 2 diabetic patients with microalbuminuria. The patients were randomly allocated to captopril (n = 9) or placebo (n = 6). After 1-year therapy no significant decrease in blood pressure was demonstrated with captopril (139 ± 17/80 ± 9 versus 138±13/76±6 mmHg) or placebo (138 ± 9/75 ± 6 versus 135 ± 14/79 ±10 mmHg). Only in a small hypertensive subgroup (n = 4) treated with captopril did we find a significant reduction in blood pressure (154 ± 2/88 ± 1 versus 142 ± 7/78 ± 5 mmHg,P < 0.05). The urinary albumin excretion rate did not change significantly either in the captopril group (95.6 mg/24 h, 25th percentile 138.4, 75th percentile 25.1; versus 127.8 mg/24 h, 25th percentile 29.3, 75th percentile 222) or in the placebo group (99.2 mg/24 h, 25th percentile 58.5, 75th percentile 125.8; versus 120.9 mg/24 h, 25th percentile 62.1, 75th percentile 179.7). There were also no alterations in renal blood flow or filtration rate. In the hypertensive subgroup treated with captopril a reduction in urinary albumin excretion rate after 3 and 6 months of treatment was observed (captopril 73.4 versus 24 and 41 mg/24 h,P < 0.05), but not after 12 months. Triglyceride and cholesterol levels remained constant before and after treatment while glycosylated hemoglobin decreased significantly after 12 months captopril (7.8 ± 0.9 versus 6.9 ± 0.7 mg%,P < 0.03). We conclude that in patients with type 2 diabetes with microalbuminuria angiotensin-converting enzyme inhibitors may have protective renal effects in so far as a lack of increase in urinary albumin excretion is equivalent to low progression in renal disease.Abbreviations ACE angiotensin-converting enzyme - GFI glomerular filtration rate - HbA glycosylated hemo globin - RPF renal plasma flow - UAER urinary albumin excretion rate Correspondence to: R. Prager     

Risk genotypes and haplotypes of the GLUT1 gene for type 2 diabetic nephropathy in the Tunisian population

Objective: Diabetic nephropathy (DN) is a long-term complication of both type 1 and type 2 diabetes. Genetic studies on DN have been of little help so far, since several genetic association studies have shown conflicting results. Here we report the findings of a case-control study on five SNPs in the glucose transporter 1 (GLUT1) gene. The study investigated the association of five GLUT1 genotypes and haplotypes with DN.

Research design and methods: All subjects, 126 DN (cases) and 273 type 2 diabetes (controls), were genotyped using the polymerase chain reaction restriction fragment length polymorphism.

Results: The TT and the AA genotypes of the Haell and Enh2 SNP1, increased the risk of DN. The study also identified CGT as the highest risk haplotype (4.4-fold) followed by CAT with an increased risk of DN of 2.6-fold.

Conclusions: The GLUT1 gene confers susceptibility to DN in type 2 diabetes patients in the Tunisian population.