醛糖还原酶在糖尿病肾病发生中的作用研究进展

醛糖还原酶(ALR)是多元醇通道重要的限速酶,功能性ALR基因定位于染色体7q35,可能由于ALR基因调控区的基因多态性,携带某种基因型的糖尿病患者,ALR产物增加,活化多元醇通道,从而参与糖尿病肾病的发生和发展。     

醛糖还原酶基因多态性与2型糖尿病肾病的相关性研究

醛糖还原酶 (ＡＲ)是多元醇通路的限速酶。近年来研究发现ＡＲ基因 5’端转录区上游 2 .1ｋｂ处的微卫星多态性与糖尿病肾病 (ＤＮ)相关。本研究探讨该多态性与中国北京汉族 2型糖尿病肾病的相关性。一、对象与方法1.对象 :糖尿病诊断按ＷＨＯ(1985 )标准 ,口服 75 ｇ葡萄糖耐量试验测定胰岛素及Ｃ肽释放曲线确定糖尿病分型 ,排除 2型糖尿病中胰岛细胞抗体 (ＩＣＡ)或谷氨酸脱羧酶(ＧＡＤ)抗体阳性者。所选病例均排除高血压病史。 (1)糖尿病肾病组 (ＤＮ ＤＲ)组 :ＤＮ诊断标准为 2 4小时尿白蛋白≥30ｍｇ持续 6月以上。排除尿路感染等其他…     

醛糖还原酶基因与糖尿病慢性并发症

讨论了醛糖还原酶基因的结构、调节及其变异与糖尿病慢性并发症之间的关系     

醛糖还原酶基因启动子区C(-106)T多态与糖尿病肾病的相关性

对2型糖尿病伴有糖尿病肾病患者（DN）70例、不伴DN患者64例和85例正常对照研究显示，醛糖还原酶启动子区C（-106）T的CC基因型是中国北方汉人发生DN的危险因子。     

醛糖还原酶基因内含子多态性与2型糖尿病肾病的相关性研究

209例2型糖尿病人和84名健康对照者的醛糖还原酶基因第8内含子第95位点AC多态性研究发现基因型AA的频度,糖尿病肾病患者是7%,明显高于非肾病组(1%)和健康对照组(1%)。这提示,该酶第8内含子基因型AA可能是糖尿病肾病的危险因子。     

黄芩甙对糖尿病患者红细胞醛糖还原酶活性及早期糖尿病肾病的影响

目的：观察中药黄芩甙对糖尿病患者醛糖还原酶（AR）活性抑制作用及对早期糖尿病肾脏病变的疗效。方法：36例早期糖尿病肾病患者随机分为对照组（18例,常规治疗）和治疗组（18例,常规治疗＋黄芩甙,疗程4w,治疗前后分别测定红细胞AR,尿白蛋白排泄率（UAER）,血,尿（β2－MG）,结果：上述指标治疗前二组相似,治疗后治疗组AR活性受到抑制,明显低于对照线,UAER,血β2－MG明显低于对照组,结果,黄苓甙可明显抑制AR活性,降低UAER,血β2－MG,有助于早期糖尿病肾病的治疗。     

蛋白激酶C和TGF-β1在糖尿病大鼠肾脏中的表达

目的研究糖尿病大鼠肾小球中蛋白激酶C（PKC）活性变化和转化生长因子-β1（TGF-β1）的动态变化，探讨二者之间相互关系及其与糖尿病肾病（DN）发生发展的关系。方法用链脲佐菌素制备糖尿病大鼠实验模型，将大鼠随机分为正常对照组（N）、糖碌病2w组（DM2）和糖尿病4w组（DM4）。断头处死，分离肾小球，提取纯化胞浆及胞膜蛋白，利用（γ-32^P）-ATP底物磷酸化的方法检测胞浆及胞膜PKC括性。用免疫组织化学和400倍光镜检测TGF-β1在各组大鼠肾脏的表达。结果（1）糖尿病大鼠肾小球细胞内总的PKC活性与对照组无显著性差异，胞浆PKC活性略有下降，相差不显著；肾小球细胞膜PKC活性明显高于正常对照组，膜结合PKC百分比显著增加，且细胞膜PKC活性与肾脏肥大指数及Cer呈正相关。（2）糖尿病组TGF-β1的表达多于正常组。（3）肾小球细胞膜PKC活性与TGF-β1的表达正相关。结论高血糖慢性刺激可引起肾小球PKC活性增高，并诱导TGF-β1的表达。在糖尿病早期肾内血流动力学改变中发挥着重要的调控作用。     

醛糖还原酶基因多态性与糖尿病微血管并发症的相关性研究

目的　探讨醛糖还原酶（ＡＲ）基因多态标记（ＡＣ）ｎ 与中国人２ 型糖尿病微血管并发症之间的关系。方法　用３２Ｐ引物末端标记法,经聚合酶链反应变性聚丙烯酰胺凝胶电泳,检测３６２ 例中国人（２１７ 人为２ 型糖尿病,其中１５６ 人合并肾病或视网膜病变,１４５ 人为非糖尿病对照）ＡＲ 基因的二核苷酸（ＡＣ）ｎ 串联重复序列多态标记,比较各组间的等位基因频率和基因型频率。结果　（１）在非糖尿病对照者和２ 型糖尿病患者中,见到七种ＡＲ 基因的等位基因,其中以（ＡＣ）２４（１３８ｂｐ） 等位基因（Ｚ）最常见。（２）同时合并肾病和视网膜病变的２ 型糖尿病组与无并发症的２ 型糖尿病组相比,Ｚ＋ ６等位基因频率显著减少（Ｆｉｓｈｅｒ 确切, Ｐ＝０ ．０３８）, 含Ｚ＋ ６ 等位基因的基因型频率也显著减少（Ｐ＝０．００４）。（３） 无并发症的２ 型糖尿病组与对照组比较其Ｚ＋ ６ 等位基因频率显著增加（ Ｐ＝ ０．０４４） 。（４）２ 型糖尿病合并肾病伴肾功能不全者与不伴有肾病的２ 型糖尿病者相比,Ｚ－ ２ 等位基因频率显著增加（ Ｐ＝ ０ ．０３９）,同时含Ｚ－ ２ 等位基因的基因型频率也呈增加趋势。结论　ＡＲ 的Ｚ－ ２ 等位基因是促进中国人２ 型糖尿病并发严     

川芎嗪对糖尿病鼠醛糖还原酶活性及视网膜细胞凋亡影响

雄性Wistar大鼠40只，随机平均分为4组，正常对照组、糖尿病组、糖尿病依帕司他（10mg／kg／d）治疗组、糖尿病川芎嗪（100mg／kg／d）治疗组，蝮腔注射链脲佐菌素（65mg／kg）诱发糖尿病。16周后，处死大鼠，分离晶体，测定组织AR活性，观察视网膜Bcl-2、Bax的表达。结果：1．与正常对照组相比，糖尿病组AR活性明显升高（P〈0．001），依帕司他治疗组、川芎嗪治疗组AR活性较糖尿病组明显降低（P〈0．01），而血糖无明显变化。2．糖尿病组视网膜Bcl-2、Bax蛋白表达明显增加，依帕司他治疗组、川芎嗪治疗组的Bcl-2、Bax蛋白表达较糖尿病组明显减少。结沦：1．AR过度激活通过促进Bcl～2、Bax蛋白的表达诱导细胞凋亡，而参与DR的发生与发展。2．ARts通过抑制醛糖还原酶活性，调节Bax、Bcl-2的表达抑制细胞凋亡，延缓DR的发展。3．川芎嗪对AR抑制作用与典型ARIs依帕司他相似，且价格低、副作用少，值得临床推荐应用。     

五龙克糖口服液对1型糖尿病肾病大鼠防治作用的研究

目的:观察中药五龙克糖口服液对实验性1型糖尿病肾病大鼠的防治效果,并初步探讨其作用机制。方法:选 用雄性Wistar大鼠,采用链脲佐菌素(65 mg／kg)一次性尾静脉注射,造成1型糖尿病动物模型,然后随机分成模 型组及五龙克糖组(各10只)。另取鼠龄、体重相匹配的雄性大鼠10只,尾静脉仅注射枸橼酸钠缓冲液,作为正常 对照组。造模成功后第2天即开始干预治疗。正常对照组及模型组给予蒸馏水灌胃,五龙克糖组给予复方五龙克糖的 提取液灌胃(7 ml／kg·d)。每天上午8～10 h灌胃一次,治疗时间为10周。观察治疗前、后血糖变化,治疗后24 h 尿微量白蛋白(MA)．血肌酐(Cr),血尿素氮(BUN)等指标;采用免疫组织化学方法,在光镜下观察肾组织中转 化生长因子β1(TGF-β1)表达情况并进行图像分析;在光镜下作肾组织形态学观察。结果:与模型组比较,五龙克 糖组空腹及餐后2 h血糖、BUN水平无明显变化(P>0．05),但血Cr、24 h尿MA明显降低(P<0．01)。五龙克糖 组肾组织TGF-β1表达较正常组强(P<0.01),但比模型组明显减弱(P<0．01)。肾脏组织形态学显示五龙克糖组 病理改变较模型组径。结论:中药五龙克糖能改善糖尿病时肾脏的病理变化,明显减少尿MA的排泄,延缓肾功能 不全的发生,其作用机制与抑制肾组织TGF-β1的表达有关。     

Polyol pathway and diabetic nephropathy revisited: Early tubular cell changes and glomerulopathy in diabetic mice overexpressing human aldose reductase

Aims/Introduction: The polyol pathway has long been involved in the pathogenesis of diabetic nephropathy. It remains still unclear, however, how the polyol pathway is implicated in this process. We explored the effects of the enhanced polyol pathway on renocortical tubular cells and glomeruli in experimentally‐induced diabetes. Materials and Methods: Transgenic mice (Tg) overexpressing human aldose reductase were made diabetic by streptozotocin and followed for 8 weeks. Renocortical pathology, expressions of tonicity‐responsive enhancer binding protein (TonEBP) and carboxymethyllysine of advanced glycation end‐products, were examined. Wild‐type non‐transgenic mice (Wt) were also made diabetic and served as controls. Results: Diabetic Tg showed augmented expression of TonEBP in renocortical tubular cells with vacuolated degenerative changes. These structural changes were associated with pronounced deposition of carboxymethyllysine. There was a significant increase in kidney weight, glomerular size, and mesangial area in diabetic animals and there was a trend for more severe changes in these measures in diabetic transgenic mice compared with those in control diabetic mice. Treatment with aldose reductase inhibitor significantly prevented polyol accumulation, mesangial expansion and expressions of TonEBP and carboxymethyllysine in diabetic Tg, but its effects on the renal structure were equivocal in control diabetic Wt. Conclusions: Our findings suggest that tubuloglomerular change might contribute to early diabetic nephropathy under the influence of the enhanced polyol pathway. (J Diabetes Invest, doi: 10.1111/j.2040‐1124.2010.00071.x , 2010)     

野黄芩甙元对糖尿病大鼠肾脏的保护作用

目的 探讨蛋白激酶C(PKC)抑制剂野黄芩甙元(EB)对糖尿病(DM)大鼠肾脏病变的保护作用及作用机制。方法 观察EB对DM大鼠肾病理、肌酐清除率(Ccr)、尿白蛋白排泄率(UAER)的影响，以及对肾组织局部内皮素(ET)、转化生长因子β1(TOF-β1)含量的影响。结果 EB可降低DM大鼠Ccr、UAER及系膜基质含量，降低肾内局部ET、TGF—β1的水平。结论 可能通过降低肾组织局部ET及TGF-β1的含量起到对糖尿病肾病(DN)的保护作用。     

血小板衍化生长因子-BB与糖尿病肾病

血小板衍化生长因子-BB(PDGF-BB)是PDGF经典的存在形式之一,其在肾脏中的作用主要包括诱导肾脏系膜细胞增生,促进细胞外基质积聚及单核-巨噬细胞在肾组织浸润。PDGF-BB在糖尿病肾病肾脏中表达增高,高水平的PDGF-BB可引起肾小球系膜细胞增生和细胞外基质积聚,亦参与肾小管及其间质改变,从而在糖尿病肾病的发生发展中起着重要作用。PDGF-BB还能通过诱导转化生长因子-β表达协同促进糖尿病肾病肾纤维化的发展。阻止PDGF-BB表达及其信号转导将有可能是糖尿病肾病的一种治疗手段。     

Genetic deficiency of aldose reductase counteracts the development of diabetic nephropathy in C57BL/6 mice

Aims/hypothesis  

The aim of the study was to investigate the effects of genetic deficiency of aldose reductase in mice on the development of key endpoints of diabetic nephropathy.     

醛糖还原酶抑制剂依帕司他改善大鼠糖尿病膀胱病变组织的功能

目的 研究醛糖还原酶(AR)、一氧化氮(NO)与糖尿病膀胱病(DCP)的关系,探讨醛糖还原酶抑制剂(ARIs)依帕司他对DCP的治疗作用. 方法 雄性SD大鼠60只分三组;正常组、糖尿病组和治疗组.测定膀胱组织AR活性,NO含量及一氧化氮合酶(NOS)活性. 结果与正常组相比,糖尿病组膀胱组织内AR活性、NO含量和NOS活性升高(P<0.01),治疗组AR活性、NO含量和NOS活性均低于糖尿病组(P<0.05). 结论 依帕司他通过抑制AR活性,调节NO含量变化及NOS活性,减轻氧化应激反应,改善膀胱组织的功能.     

An updated meta-analysis of methylenetetrahydrofolate reductase gene 677C/T polymorphism with diabetic nephropathy and diabetic retinopathy

Studies investigating the association of methylenetetrahydrofolate reductase (MTHFR) gene 677C/T polymorphism with diabetic nephropathy and diabetic retinopathy have so far reported inconclusive results. We therefore aim to address this inconclusiveness by conducting a meta-analysis. Random-effects model was applied irrespective of between-study heterogeneity. Data and study quality were assessed in duplicate. A total of 7807 and 1599 subjects from 21 and 8 studies were analyzed for diabetic nephropathy and diabetic retinopathy, respectively. Carriers of 677TT genotype were 1.71 (95% confidence interval [95% CI]: 1.02-2.88; P = 0.042) and 2.89 (95% CI: 1.51-5.53; P = 0.001) times more likely to develop diabetic nephropathy separately relative to diabetic patients without nephropathy and nondiabetic controls. Likewise, this association was preserved for diabetic patients with retinopathy referring to those without (odds ratio [OR] = 1.86; 95% CI: 1.21-2.86; P = 0.004). Subgroup analyses showed that ethnicity was a possible confounder, especially in West Asians and Africans, and so were gender and duration of diabetes mellitus in diabetic nephropathy studies. Probability of publication bias was low across all comparisons as reflected by the funnel plot and corresponding test. Taken together, our results demonstrate that MTHFR gene 677TT genotype might confer a moderately augmented risk for diabetic nephropathy and diabetic retinopathy.     

依那普利及波生坦对糖尿病肾病大鼠干预作用的初步研究

目的　探讨依那普利和波生坦对糖尿病肾病 (DN)的治疗作用。　方法　Wistar大鼠摘除右肾后 ,静脉注射链脲佐菌素致成糖尿病模型鼠 ,分为 4组 :糖尿病模型 (Veh)组 (给缓冲液 ) ;波生坦干预 (Bos)组 (10 0mg·kg-1·d-1) ;依那普利干预 (Ena)组 (10mg·kg-1·d-1) ;波生坦及依那普利联合干预 (Bos Ena)组 (两药剂量同上 )。尚设单肾对照组 (静脉注射缓冲液 )。每组大鼠 6只。 2 0周时测量体重、血压、血糖及 2 4h尿蛋白定量 ,处死大鼠 ,取左肾称重 ,用RT PCR及免疫组化法检测肾组织中Ⅰ型胶原 (CⅠ)、Ⅳ型胶原 (CⅣ)、转化生长因子 β1(TGF β1)、纤溶酶原激活剂抑制物 1(PAI 1)及金属蛋白酶组织抑制物 1(TIMP 1)的mRNA及蛋白质表达。　结果　与对照组相比 ,Veh组大鼠平均动脉压、血糖、尿蛋白定量及肾重 /体重比值均显著上升 (P <0 0 5 ) ,肾组织的CⅠ 、CⅣ 、TGF β1、PAI 1及TIMP 1的mRNA及蛋白质表达均显著上调 (P <0 0 5 )。Bos、Ena及两药联合治疗后 ,上述上调指标除血糖外均显著抑制 (P <0 0 5 ) ,而 3组间抑制率差异无显著意义 (P >0 0 5 )。　结论　无论非选择性内皮素 1受体拮抗剂Bos或血管紧张素转换酶抑制剂Ena,对DN均具有肯定疗效 ;两药可能通过下调肾组织中TGF β1、PAI 1及TIMP 1表     

蛋白激酶C-α、蛋白激酶C-βⅠ和蛋白激酶C-βⅡ在糖尿病肾病患者肾小球中的表达及其意义

目的探讨蛋白激酶C（PKC）-α、PKC-βⅠPKC-βⅡ在人正常肾组织以及糖尿病肾病患者（DN）肾小球中的表达及其与肾小球内转化生长因子（TGF-βⅠ和血管内皮细胞生长因子（VEGF）的关系。方法以正常肾组织为对照，取糖尿病肾病患者活检肾组织，采用PAS染色判定肾小球细胞外基质（ECM）扩张程度，半定量免疫组织化学法检测PKC-α、PKC-βⅠPKC-βⅡ、TGF-βⅠVEGF在肾小球中的表达。结果（1）PKC-α、PKC-βⅠPKC-βⅡ在正常肾小球中均有表达；（2）糖尿病肾病患者肾小球中PKC-α、PKC-βⅠTGF-β1和VEGF的表达明显增强而PKC-βⅡ的表达明显减弱，且肾小球ECM明显增加。其中PKC-α与VEGF的表达变化呈正相关（r，=0．600，P=0．039），PKC-βⅠTGF-βⅠ表达呈正相关（r=0．521,P=0．043）。结论PKC-α、PKC-βⅠPKC-βⅡ在正常人以及DN患者肾小球中的表达不同；PKC-α可能通过增强肾小球内VEGF的表达，而PKC-βⅠ能通过增强肾小球内TGF-βⅠ表达参与DN的发病过程。     

Aldose reductase gene polymorphisms and susceptibility to diabetic nephropathy in Type 1 diabetes mellitus.

AIMS: To investigate association and linkage between DNA sequence variants in the aldose reductase (AR) gene on chromosome 7q35 and diabetic nephropathy (DN) in Type 1 diabetes mellitus. METHODS: By sequencing the promoter region and 10 exons in eight DN cases and eight controls, a frequent bi-allelic polymorphism (C-106T) was discovered. This polymorphism and the known 5'ALR2 dinucleotide repeat polymorphism were genotyped in unrelated cases with advanced nephropathy (n = 221) and unrelated controls with normoalbuminuria (n = 193). For a family based study, 166 case-trios (case and both parents) and 83 control-trios (control and both parents) were also genotyped. RESULTS: In the case-control study, carriers of the Z-2 allele of the 5'ALR2 polymorphism had a significantly higher risk of DN than non-carriers (odds ratios: 1.6 for heterozygotes and 2.1 for homozygotes, P<0.05 for each). The same was true for carriers of the T allele of the C-106T polymorphism (odds ratios: 1.6 for heterozygotes and 1.9 for homozygotes, P<0.05 for each). Moreover, the haplotype carrying both risk alleles was in excess in DN cases. In the family study, transmission of risk alleles from heterozygous parents was consistent with the case-control study, excess transmission in case-trios and deficient in control-trios. CONCLUSIONS: Association between DN and two DNA sequence variants in the promoter region of the AR gene implicates the polyol pathway in the development of kidney complications in Type 1 diabetes mellitus. Further examination of the molecular mechanisms underlying these findings may provide insight into the pathogenesis of DN.