白藜芦醇对糖尿病肾损害大鼠超微结构的影响

目的观察白藜芦醇(Res)对糖尿病肾病大鼠模型超微结构的保护作用及可能机制。方法 Wistar大鼠用链脲佐菌素腹腔注射诱导糖尿病,随机分为正常对照(NC)、糖尿病及Res组,腹腔给药10 w后观察各组肾功能及肾脏超微结构的变化,检测大鼠血浆及肾组织氧化、抗氧化指标。结果与NC组比较,糖尿病大鼠肾小球体积(MGV)增加、基底膜(GBMT)增厚,伴尿蛋白排泄量和肾脏指数升高,血尿素氮(BUN)和肌酐(Cr)升高(P均<0.01),血浆和肾皮质丙二醛(MDA)含量明显增加,总抗氧化力(TAC)和巯基(SH)水平显著降低(P均<0.01);Res明显降低糖尿病大鼠尿蛋白排泄量、肾脏指数、MGV、GBMT、BUN和Cr水平,减少血浆和肾皮质MDA含量,提高TAC和SH水平(P<0.05,P<0.01)。结论 Res对糖尿病大鼠的肾损害具有保护作用,能明显改善肾损害模型的超微结构,其机制可能与抗氧化作用有关。     

伊贝沙坦对糖尿病大鼠一氧化氮系统及肾脏的影响

目的　研究伊贝沙坦对糖尿病大鼠一氧化氮 (NO)系统及肾脏的影响。　方法　随机将 4 0只Wistar大鼠分为 4组 ,每组 10只 ,分别为正常对照组、糖尿病组、伊贝沙坦组和开搏通组。病程 12周时处死大鼠 ,取血、尿和肾脏标本 ,测定尿量、体重、肾重 /体重比值、血糖、糖化血红蛋白(HbA1c) ;测定血清、尿液和肾组织的NO水平 ,通过免疫组化方法 ,检测肾脏组织诱导型一氧化氮合酶 (iNOS)蛋白的合成 ,并通过光镜和电镜观察肾脏的病理结构变化。　结果　治疗 12周后 ,糖尿病各组大鼠的尿量、肾重 /体重比值、血糖、HbA1c、血清、尿液和肾脏组织的NO水平、肾脏组织iNOS蛋白的合成、肾小球体积和肾小球基底膜厚度明显高于或大于正常组 ,体重明显低于正常组 (P <0 .0 1) ;伊贝沙坦组大鼠的血清、尿液和肾脏组织的NO水平、肾脏组织iNOS蛋白的合成、肾小球体积和肾小球基底膜厚度比糖尿病组明显减少 (P <0 .0 5 ) ;血清、尿液和肾组织NO水平与肾小球体积和肾小球基底膜厚度呈正相关。　结论　伊贝沙坦能延缓糖尿病大鼠肾脏功能损害的进展 ,其机制可能与伊贝沙坦不同程度地抑制糖尿病大鼠NO的产生有关。     

金属蛋白酶-2和金属蛋白酶组织抑制物-1与糖尿病肾病关系的实验研究

目的　观察链脲佐菌素 (STZ)实验性糖尿病大鼠肾脏基质金属蛋白酶 2 (MMP 2 )及金属蛋白酶组织抑制物 1(TIMP 1)蛋白的表达及功能、形态学改变 ,探讨MMP 2、TIMP 1在糖尿病肾病(DN)发生机制中的意义。　方法　 2 0只雄性Wistar大鼠随机分为糖尿病组和正常对照组 ,分别于第1、2、4、6、8周测定尿白蛋白排泄率 (UAER) ,第 9周用Western印迹方法检测MMP 2、TIMP 1蛋白表达水平 ,电镜观察肾小球基底膜厚度 (GBMT)。　结果　糖尿病组较正常组TIMP 1蛋白表达明显增加 ,MMP 2蛋白表达显著降低 (P <0 .0 1)。 4、6、8周UAER显著增加 ,GBMT明显增厚 (P <0 .0 1)。电镜发现糖尿病组肾小球基底膜弥慢性增厚 ,局部有系膜细胞插入和双轨征。　结论　持续高血糖可使大鼠肾脏MMP 2下降 ,TIMP 1增加。导致肾小球基底膜增厚 ,UAER增加。MMP 2、TIMP 1失衡可能对糖尿病肾病功能和形态学改变有重要意义。     

老年人非肾脏疾病尿蛋白／肌酐比值检测及其意义

目的采用一次尿蛋白／肌酐比值(Pr／Cr比值)检测老年高血压病，糖尿病和冠心病患尿微量置白以诊断早期肾脏损害。方法收集8～12时任意尿标本，采用考马斯亮蓝(CBG)染料综合比色法测定尿微量置白；尿肌酐采用酸性苦味酸法测定。结果本组尿蛋白定性试验阴性的老年高血压病、冠心病和糖尿病患尿Pr／Cr比值较老年健康对照组明显升高(P<0．01)。结论尿Pr／Cr比值可以作为老年糖尿病、高血压病和冠心病患肾脏早期损害的诊断指标。     

糖尿病大鼠肾脏一氧化氮变化与肾小球硬化及非酶糖化的关系

肾小球硬化为糖尿病肾病 (DN)的病理改变 ,其基本病变为肾小球基底膜增厚和系膜基质的增生。糖尿病 (DM)肾组织胶原蛋白非酶糖化是 DM肾小球硬化重要的原因之一 [1 ,2 ]。有研究表明一氧化氮 (NO)具有抑制肾小球系膜细胞外基质积聚的作用 ,而蛋白非酶糖化致糖基化终产物 (AGE)的大量形成则阻碍了 NO的抗细胞增殖作用 [3 ]。本实验通过观察链脲佐菌素(STZ)诱导的 DM大鼠在不同病程肾组织 NO含量和一氧化氮合酶 (NOS)活性、AGE、胶原含量及肾组织形态学变化 ,探讨DN发病的相关机制。材料和方法1.材料 :选择体重 15 0～ 2 0 0 g的…     

2型糖尿病不同病程与胰岛功能及肾脏功能的相关研究

目的探讨2型糖尿病不同病程与胰岛素功能及肾脏功能的关系。方法回顾分析该院2019年1月—2020年1月收治的120例胰岛素泵治疗的2型糖尿病患者,依据病程分成4组:1年(A组)、1～5年(B组)、5～10年(C组)及10年(D组),观察各组患者临床特点、胰岛功能及肾脏功能指标,并分析病程与胰岛功能和肾脏功能的相关性。结果各组患者空腹血糖(FPG)、尿素氮(BUN)差异无统计学意义(P0.05);A组糖化血红蛋白(HbA1c)、胰岛素抵抗指数(Homa-IR)较B、C、D组显著升高,差异有统计学意义(P0.05);D组Homa-β、肾小球滤过率(eGFR)较A、B组显著降低,差异有统计学意义(P0.05),D组(Cr)肌酐、β2微球蛋白(β2-MG)、尿蛋白肌酐比值(ACR)较A、B组显著升高,差异有统计学意义(P0.05)。Pearson相关分析显示,病程与Cr、β2-MG及ACR呈显著正相关(P0.05);与eGFR、Homa-β及Homa-IR呈显著负相关(P0.05)。结论 2型糖尿病患者病程第1年胰岛素抵抗为主;病程第5年,肾脏功能显著受损;病程第10年,胰岛功能显著性下降,为临床及时干预,延缓病情进展提供理论依据。     

HbA1c与mALB／Cr的相关性研究

对216例DM患者FPG、PPG、HbA1c和UMA／Cr进行测定、分析并与对照组比较。结果DM患者的FPG、PPG、HbA1c和UMA／Cr明显高于对照组（P〈0．01），且HbA1c和UMWA／Cr成正相关（r=0．522，P〈0．01）。结论DM患者HbA1c和UMA／Cr水平与DM早期肾脏微血管改变密切相关。     

长链非编码RNA Blnc1在糖尿病肾脏疾病患者血清中的表达及临床意义

目的 探究长链非编码RNA(LncRNA)Blnc1在糖尿病肾脏疾病(DKD)患者血清中表达及其临床意义。方法 纳入我院56例DKD患者作为DKD组，50例单纯2型糖尿病(T2DM)患者作为T2DM组，56例同期体检健康者作为对照组。比较各组受试者一般临床资料、实验室检查结果及血清LncRNA Blnc1表达水平。采用Pearson相关分析探讨血清LncRNA Blnc1表达水平与临床指标的相关性。采用多因素logistic回归分析探讨T2DM患者发生DKD的危险因素。采用受试者工作特征(ROC)曲线分析血清LncRNA Blnc1对DKD的诊断价值。结果 T2DM组和DKD组患者血肌酐(SCr)、糖化血红蛋白(HbA1c)水平及血清LncRNA Blnc1表达水平均高于对照组(P<0.05)。DKD组患者SCr、HbA1c、尿白蛋白/肌酐比值(UACR)、24 h尿蛋白水平、糖尿病病程及血清LncRNA Blnc1表达水平均明显高于T2DM组，估算的肾小球滤过率(eGFR)低于T2DM组(P<0.05)。多因素logistic回归结果显示，糖尿病病程、UACR、HbA1c...     

二甲双胍对2型糖尿病大鼠肾组织单核细胞趋化蛋白1表达影响的观察

目的观察二甲双胍(MET)对T2DM大鼠肾组织单核细胞趋化蛋白1(MCP-1)表达的影响及对DKD肾损害的保护机制。方法高脂饲料喂养结合腹腔注射小剂量STZ建立T2DM大鼠模型,30只造模成功大鼠随机分为T2DM组、MET组[300 mg/(kg·d)]和格列本脲干预组[GLY,5 mg/(kg·d)],每组各10只,并设正常对照组(NC,n=10)。检测各组FPG、HbA_1c、FIns、BUN、UACR、血清MCP-1水平及肾小球基底膜厚度(GBMT),检测肾组织MCP-1蛋白及mRNA表达情况。结果与NC组比较,T2DM组FPG、HbA_1c、MCP-1 mRNA、MCP-1_(IOD)、GBMT和血清MCP-1升高(P0.05)。与T2DM、GLY组比较,MET组MCP-1 mRNA、MCP-1_(IOD)、GBMT、血清MCP-1降低(P0.05)。结论 MET可下调T2DM大鼠肾组织MCP-1表达及血清MCP-1水平,可能与其肾脏保护作用有关。     

HbA1c与mALB／Cr的相关性研究

对216例DM患者FPG、PPG、HbA1c和UMA／Cr进行测定、分析并与对照组比较。结果DM患者的FPG、PPG、HbA1c和UMA／Cr明显高于对照组（P〈0．01），且HbA1c和UMWA／Cr成正相关（r=0．522，P〈0．01）。结论DM患者HbA1c和UMA／Cr水平与DM早期肾脏微血管改变密切相关。     

Localization of advanced glycation endproducts in the kidney of experimental diabetic rats

Advanced glycation endproducts (AGE) have been proposed as a major mediator in the development of various diabetic complications. In order to evaluate the involvement of AGE in the development of diabetic nephropathy, we examined the localization of AGE in the kidney of the streptozotocin-induced diabetic rats immunohistochemically using a monoclonal antibody directed to AGE. In the diabetic rats, glomerular hypertrophy, thickening of the glomerular basement membrane, and expansion of mesangial matrix were observed. AGE was detected in expanded mesangial area and glomerular basement membrane in the kidneys of diabetic rats. The present results suggest that AGE may participate in the development of diabetic nephropathy.     

Prevention and treatment of diabetic nephropathy

Increasing number of diabetic patients develop different stages of renal failure. However, often an inappropriate parameter, the serum creatinine is measured as a marker of glomerular function. Calculated glomerular filtration rate or endogenous creatinine clearance are suggested to be used for the estimation of the glomerular function. Important structures preventing proteinuria in the kidney are glomerular basement membrane, podocytes and proximal tubular cells. In diabetes mellitus loss of nephrin of podocytes can play a role in the development of microalbuminuria, and podocyte desquamation may result in the progression to proteinuria. In diabetes mellitus there is an increased formation of advanced glycation endproducts (AGE), of which the only elimination organ is the kidney. The AGE induce proteinuria and atherosclerosis. Therefore, in diabetes mellitus a vicious circle develops due to proteinuria, nephron loss and accumulation of AGE, which play a role in the initiation and progression of diabetic nephropathy and atherosclerosis. Angiotensin converting enzyme inhibitors and angiotensin receptor blockers having antiproteinuric effect may decrease the risk of diabetic nephropathy and atherosclerosis. Improvement of carbohydrate metabolism with a consequential decrease in the formation of AGE is an important contributor to the prevention and treatment of diabetic nephropathy and atherosclerosis.     

伊贝沙坦对大鼠糖尿病模型肾脏肥大和肾小球毛细血管基底膜厚度的影响

目的 观察伊贝沙坦对大鼠糖尿病模型肾脏肥大和肾小球毛细血管基底膜厚度的影响。方法 将SD大鼠分为糖尿病肾病组(A组)、伊贝沙坦治疗组(B组)、健康对照组(C组),A和B组大鼠制成糖尿病模型,B组予以50mg／kg伊贝沙坦灌胃。观察第4、8、12周大鼠的血糖、体重、尿白蛋白、24h尿蛋白的改变及第12周时的肌酐清除率(Ccr)、肾重、肾脏肥大指数、肾组织总蛋白含量、肾小球面积和体积、肾小球毛细血管基底膜(GBM)厚度的改变。通过免疫组化观察肾结缔组织生长因子(CTGF)和转化生长因子(TGF)-βl的表达。结果 A组和B组大鼠血糖较C组明显升高且维持在一个较高水平(P＜0．01)。A组大鼠的体重较C组明显下降,B组有所增加(P＜0．05)。随时间的推移(第4、8、12周),A组大鼠尿蛋白、尿白蛋白逐渐增加,B组明显减少(P＜0．01)。A组大鼠Ccr较C组显著升高(P＜0．01),B组Ccr明显下降(P＜0．05)。至第12周时,A组大鼠肾脏重量、肾脏肥大指数、肾组织总蛋白含量、肾小球面积和体积均较C组明显增加(P＜0．01),B组均较A组降低(P＜0．01,P＜0．05)。免疫组化半定量分析显示,A组大鼠CTGF与TGF-βl的表达均高于C组(P＜0．01),B组明显低于A组(P＜0．01,P＜0．05)。A组大鼠GBM较C组明显增厚(P＜0．01),B组较A组明显变薄(P＜0．01）。肾小球CTGF、TGF—βl的表达与肾脏体积呈正相关(r＝0．83,r＝0．83;P＜0．05)。结论 早期应用伊贝沙坦可抑制糖尿病大鼠早期肾脏肥大和CTGF表达等。     

糖尿病肾病与颈动脉内膜中层厚度相关性研究

探讨糖尿病肾病的进展与大动脉硬化的关系.结果 发现糖尿病患者颈动脉内膜中层厚度 (CIMT)与尿白蛋白/肌酐显著正相关,与肾小球滤过率显著负相关.提示有微量白蛋白尿的糖尿病患者,CIMT的增厚预示其糖尿病临床肾病发生的风险增加.     

艾塞那肽对糖尿病肾病小鼠足细胞的保护作用

目的探讨艾塞那肽对糖尿病肾病小鼠足细胞的作用。方法通过给予C57BL/6J小鼠高脂饮食并注射链脲佐菌素建立糖尿病肾病模型,按随机数字表法将其分为糖尿病肾病对照组(DN组,n=8)、艾塞那肽干预组(DN+Ex组,n=8)。同时将普通饲料喂养的C57BL/6J小鼠作为正常对照组(NC组,n=8)。干预结束后,测定血糖、肾功能和尿微量白蛋白/肌酐比值,采用过碘酸希夫染色(PAS)观察小鼠肾小球病理学改变,实时定量PCR分析肾小球组织中促纤维化分子Ⅳ型胶原蛋白(Collagen Ⅳ)、转化生长因子-β(TGF-β)和纤维连接蛋白(Fibronectin)基因转录水平,免疫荧光染色及电镜观察足细胞损伤及凋亡情况,Western印迹法检测肾小球组织中裂孔膜肾病蛋白(Nephrin)、活化型半胱氨酸天冬氨酸蛋白酶3(Cleaved caspase-3)、蛋白激酶B(Akt)和磷酸化Akt(p-Akt)表达水平。结果与DN组相比,DN+Ex组小鼠尿微量白蛋白/肌酐比值显著降低(P<0.01)。PAS染色及分析发现,艾塞那肽干预治疗改善了糖尿病肾病小鼠的肾小球系膜基质增生和肾小球肥大(P<0.05)。实时定量PCR显示,与DN组小鼠相比,DN+Ex组小鼠的肾小球组织中Collagen Ⅳ、TGF-β和Fibronectin基因表达下调(P<0.01)。免疫荧光染色和电镜显示,艾塞那肽干预治疗改善了糖尿病肾病小鼠的足细胞损伤及凋亡。Western印迹法发现,艾塞那肽干预后糖尿病小鼠肾小球组织中的Nephrin水平升高(P<0.01)、Cleaved caspase-3水平下降(P<0.01)、p-Akt水平升高(P<0.01)。结论艾塞那肽能够改善糖尿病肾病小鼠的足细胞损伤及凋亡,减少蛋白尿,从而延缓糖尿病肾病的进展。这种保护作用可能与激活肾小球中磷酸肌醇3-激酶(PI3K)/Akt信号通路有关。     

Urinary excretion of advanced glycation endproducts in patients with type 2 diabetes and various stages of proteinuria

OBJECTIVE: The objective of the study was to detect AGE-immunoreactive proteins in urine, and to evaluate AGE excretion at various stages of diabetic nephropathy in type 2 diabetes assessed by the level of proteinuria. METHODS: AGEs were measured in 24-h urine collection of patients with normoalbuminuria (N) (n=22), microalbuminuria (Mi) (n=31), macroalbuminuria (Ma) (n=28), and overt proteinuria with elevated serum creatinine level (PC) (n=25). A competitive ELISA with polyclonal anti-AGE antibodies was used to monitor AGE excretion. RESULTS: Multiple comparison of urine AGE content among various stages of proteinuria showed significant differences (summary p<0.000). Fifty percent of samples from the group of normoalbuminuric, and only 15% of samples from the group of microalbuminuria patients were AGE negative. However, there was no significant difference in AGE excretion between the patients with persistent proteinuria and elevated serum creatinine, and those with macroalbuminuria (PC vs Ma, p=0.265). None of the samples from these two groups of patients with highest AGE content in 24-h urine was negative for AGE-immunoreactivity. In addition, the ratio between 24-h urinary AGEs and urinary albumin excretion was calculated to determine whether total 24-h urinary AGE content is an index of the toxic form of albumin released in the course of diabetic nephropathy. The ratio values were log-transformed and bivariate comparison showed significant differences between the N vs Mi (p=0.006) and Mi vs Ma (p=0.000) groups. However, there was no significant difference (p=0.407) between values in the Ma and PC groups of patients. Multiple stepwise regression analysis indicated a relationship of urinary AGE-immunoreactivity with creatinine clearance values (r=0.52, p<0.001). CONCLUSION: The study demonstrated the presence of AGE-immunoreactivity in the urine of diabetic patients with various stages of proteinuria. Study results pointed to creatinine clearance as the main predictor of AGE excretion. Therefore, the measurement of urinary AGE appears to offer limited extra information in patients with impaired renal function.     

Expression and effect of TLR4 in rats with diabetic nephropathy

ObjectiveTo observe the expression of TLR4 in kidney tissue of rats with diabetic nephropathy and discuss the role of TLR4 in the occurrence and development of the diabetic nephropathy.MethodsA total of 60 clean male SD rats were selected and randomly divided into the modeling group and control group after 1 week of breeding, including 30 rats in each group. Biochemical indices as well as the protein expression of TLR4 were observed and compared between two groups at 2 w, 4 w, 6 w, 8 w and 12 w after the modeling, and the correlation between TLR4 and each biochemical indexes was analyzed.ResultsRats in the modeling group had higher levels of blood glucose, 24-hour urine protein and blood urea nitrogen after the modeling, and showed the increase in the serum creatinine, kidney/body weight ratio, CRP and serum TNF-α at 4w after the modeling, with the significant difference compared to results of the control group (P<0.05). The cross-section area and mean volume of glomerulus in the modeling group at 4 w, 6 w, 8 w and 12 w were significantly higher than those in the control group, with the statistically significant difference (P<0.05). The expression of TLR4 at each time point in the control group was relatively low. Rats in the modeling group had the high expression of TLR4 in kidney's glomerular basement membrane, proximal convoluted tubule and renal interstitial area since 2 w, with the significant difference compared to the control group (P<0.05). The expression in rats of the modeling group was higher than the one of the control group since the 2nd week. As the time flied, its expression increased, with the statistically significant difference between two groups (P<0.05). There was certain correlation between the protein expression of TLR4 and the increased serum titer of 24-hour urine protein excretion, serum creatinine, CRP and TNF-α.ConclusionsTLR4 may activate the immuno-inflammatory reactions to play a role in the occurrence and development of the diabetic nephropathy.     

Serum levels of non-carboxymethyllysine advanced glycation endproducts are correlated to severity of microvascular complications in patients with Type 1 diabetes

We investigated whether serum levels of N-(carboxymethyl)lysine (CML), non-CML advanced glycation endproducts (AGEs), or pentosidine are associated with severity of diabetic microvascular complications in patients with Type 1 diabetes. Serum levels of CML, non-CML AGE, and pentosidine were measured by an enzyme-linked immunosorbent assay in 38 males and 47 females aged 31+/-8 years (mean+/-S.D.) with Type 1 diabetes for 18.7+/-7.0 years. There was a significant correlation between serum levels of CML or non-CML AGE and current HbA(1c) level (P<.01 and P<.05, respectively). The serum levels of non-CML AGE, but not CML or pentosidine, were significantly increased as normal renal status advanced to microalbuminuria, clinical nephropathy, and hemodialysis (P<.0001) and were positively correlated with urinary albumin excretion (UAE) in patients with Type 1 diabetes (P<.0001). A significant elevation of serum non-CML AGE was found in association with the severity of diabetic retinopathy (P<.0001). We found in the present study that CML levels were also increased in the stage of simple retinopathy, the early stage of clinically evident retinopathy (P<.05). Serum levels of non-CML AGE were significantly associated with the severity of diabetic nephropathy and retinopathy, suggesting a role of non-CML AGE in the progression of microvascular complications in patients with Type 1 diabetes. Since serum levels of CML were significantly increased in patients with simple retinopathy, CML may participate in the initiation of diabetic retinopathy.     

糖代谢终产物与糖尿病肾病尿毒症的相关研究

目的　研究糖代谢终产物 (AGE)水平与糖尿病肾病 (DN)尿毒症的关系 ,探讨 AGE对 DN的影响及其作用机制。方法　将 6 2例糖尿病 (DM)患者分为 DM无肾病组、DN组 (尿毒症透析组和非透析组 ) ,并设 30例体检正常者做对照组 ,采用 EL ISA法进行血清 AGE检测。结果　 DM无肾病组与对照组比较 AGE明显增高(P<0 .0 5 ) ,与血糖呈正相关 ;DN组 AGE明显增高 (P<0 .0 5 ) ,与血糖及血肌酐呈正相关。尿素症透析组与非透析组 AGE无显著性差异 (P>0 .0 5 )。结论　高血糖状态是引起 DM慢性并发症的主要原因 ;且血糖增高程度与并发症发生率直接相关 ;肾功能衰竭时 AGE明显升高 ,对全身器官造成损害。