糖尿病肾病大鼠肾脏cbfal表达与钙化的关系

目的：探讨糖尿病肾病大鼠肾脏钙化与骨分化核心结合因子1（cbfal）表达的关系。方法：用链脲佐茵素诱导糖尿病肾病大鼠模型，并以正常大鼠作为对照组，于第3、6月末分别采用VonKossa染色观察肾脏钙盐沉积、肾脏免疫组化、RT—PCR、WesterBlot法观察肾脏cbfal的表达水平。结果：糖尿病肾病大鼠肾脏有cbfal的表达，且与钙盐沉积程度有关。正常大鼠肾脏没有发现cbfal的表达。结论：糖尿病肾病晚期时肾脏出现钙盐沉积，肾脏cbfal的表达水平与钙盐沉积程度呈正相关。     

益肾方与厄贝沙坦对糖尿病肾病大鼠肾脏尾加压素Ⅱ表达及病理改变的影响

目的:探讨益肾方与厄贝沙坦对糖尿病肾病(DN)大鼠肾组织中尾加压素Ⅱ(urotensin Ⅱ,UⅡ)表达和肾脏病理改变的影响及其意义.方法:将SD大鼠以链脲佐菌素(STZ)制作糖尿病肾病大鼠模型后随机分为糖尿病肾病组(D组)、益肾方治疗组(DZ组)、厄贝沙坦治疗组(DI组),并设正常对照组.治疗8周后取肾组织,免疫组化方法检测肾组织中UⅡ表达水平,光镜及电镜观察肾脏组织结构改变.结果:与正常对照组相比,糖尿病肾病组肾组织中UⅡ表达增高,肾小球肥大,基质增多,足突融合,基底膜增厚;肾组织UⅡ表达与肾小球平均体积及基底膜厚度的关系均呈正相关.益肾方和厄贝沙坦治疗组肾组织UⅡ表达均明显减少,肾脏病理显著改善.结论:益肾方和厄贝沙坦均能减少糖尿病大鼠肾组织UⅡ的表达和改善肾脏病理改变,其保护作用可能与减少UⅡ的产生有关.     

糖尿病肾病大鼠足细胞自噬的变化

目的:研究糖尿病肾病大鼠足细胞自噬标志物LC3、P62蛋白表达的改变,探讨糖尿病肾病可能的发病机制。方法:将清洁级雄性SD大鼠20只随机分为正常对照组(n=10只)、造模组(n=10只)。采用高糖高脂饲料结合一次性腹腔注射链脲佐菌素制备糖尿病动物模型。造模成功后于第12周结束时留取大鼠尿标本,检测大鼠尿微量白蛋白、尿肌酐,摘取肾脏称量重量,分别计算尿蛋白肌酐比和肾重体重比;HE染色、PAS染色观察肾脏组织病理变化;Western-blot检测肾小球LC3、P62蛋白表达。结果:与正常对照组相比,尿蛋白肌酐比、肾重体重比在造模组均显著增加(P<0.05);P62蛋白表达亦较正常组表达显著增加(P<0.05);造模组大鼠肾脏病理在光镜下可见明显的系膜增生、基质增多。结论:糖尿病肾病大鼠肾脏足细胞自噬较正常组减低,可能是糖尿病肾病发病的机制之一。     

普罗布考对糖尿病大鼠肾组织Smads蛋白表达的影响

目的：观察链脲佐菌素（STZ）造模的糖尿病大鼠以及普罗布考（之乐）干预的糖尿病大鼠肾脏组织中Smad2、Smad4、Smad7蛋白的表达,探讨普罗布考在糖尿病肾病中的作用机制。方法：36只6周龄雄性SD大鼠（清洁级）,将其随机分为3组（对照组、糖尿病模型组、糖尿病之乐治疗组）,12周末将其处死,检测各组大鼠血糖（GLU）、血尿素氮（BUN）、肌酐（Scr）、血三酰甘油（TG）,以免疫组化的方法观察各组大鼠肾脏Smad2、Smad4、Smad7蛋白的表达。结果：糖尿病模型组、糖尿病之乐治疗组GLU、BUN、Scr较正常组明显升高,糖尿病之乐治疗组TG较正常组下降;糖尿病模型组的Smad4、Smad7在肾脏组织的表达较正常组明显增高,糖尿病之乐治疗组Smad4、Smad7在肾脏组织的表达不明显,Smad2在各组间相比差异无统计学意义。STZ诱导的糖尿病大鼠出现了明显的糖尿病肾病的病理变化,糖尿病之乐治疗组也有明显的糖尿病肾病的病理变化,但较糖尿病模型组轻。结论：TGF-β1/Smads信号通路在糖尿病肾病时是激活的,普罗布考可以通过影响糖尿病大鼠肾组织中TGF-β1/Smads信号通路中的TGF-β1、Smad2、Smad4、Smad7蛋白的表达而起到抑制纤维化的作用,延缓糖尿病肾病的进展。     

糖肾安防治早期糖尿病肾病的实验研究

目的:研究糖肾安对早期糖尿病肾病大鼠的治疗作用并探讨其可能机制。方法:采用单肾切除、高糖高脂饲料喂养、小剂量STZ(35mg/kg)腹腔注射"三联"方法建立糖尿病肾病大鼠模型。随机分为正常对照组、模型对照组、糖肾安组、福辛普利组、联合干预组,每组9只。用药6周后观察各组大鼠体重、肾重、血糖、24h尿蛋白定量、血浆Ⅳ型胶原蛋白(ColⅣ)水平的差异及肾脏病理改变、肾脏转化生长因子-β1(TGF-β1)、血管内皮生长因子(VEGF)mRNA的表达。结果:模型组大鼠体重、肾重、肾重/体重、血糖、24h尿蛋白定量及血浆ColⅣ水平、肾脏TGF-β1、VEGFmRNA表达与正常对照组比较,差异均有统计学意义(P<0.01);各用药组上述指标有所改善,与模型组比较差异均有统计学意义(P<0.05)。糖肾安组与福辛普利组比较上述指标差异均没有统计学意义。肾脏病理变化以模型组最为明显,系膜细胞重度增生,各用药组均有不同程度减轻。结论:糖肾安对早期糖尿病肾病具有良好的防治作用,其效果与福辛普利相当,中西医结合治疗能取得更好的效果。其肾保护作用可能是通过影响肾脏促纤维化生长因子TGF-β1、VEGF表达实现的。     

糖尿病大鼠肾脏中vWF的表达及丹红注射液的干预研究

目的 观察丹红注射液干预前后糖尿病大鼠血清血管性假血友病因子（vWF）水平及肾组织中vWF的表达变化,探讨糖尿病肾病发病机制及丹红的干预效果.方法 36只雄性SD大鼠随机分为3组：正常对照组10只、糖尿病组14只、丹红干预组12只.除正常对照组外,其余各组均按55mg/kg剂量一次性腹腔注射1％链脲佐菌素（STZ）建立糖尿病大鼠模型.72h后采尾静脉血测血糖,以血糖值＞16.7mmol/L作为糖尿病大鼠模型建立的标准.造模成功后,丹红干预组按2mL/kg.d剂量腹腔注射丹红注射液.干预为期8周.于第8周末,收集各组大鼠血液、尿液及肾组织标本.然后检测血糖（BG）、总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）、血肌酐（Scr）、尿素氮（BUN）.放射免疫法检测24h尿白蛋白定量.酶联免疫分析法（ELISA）测定各组大鼠血清vWF的水平.肾组织进行HE染色在光镜观察肾脏形态学变化.用免疫组化的方法检测各组大鼠肾脏vWF的表达情况.应用实时荧光定量PCR技术检测肾组织vWF的表达情况.结果 糖尿病组大鼠血糖、胆固醇、甘油三酯、低密度脂蛋白胆固醇、血肌酐、尿素氮、24h尿白蛋白定量、肾脏肥大指数、血vWF水平明显高于正常对照组（P＜0.01）,糖尿病大鼠肾脏出现病理改变,糖尿病大鼠肾组织vWF的表达较正常对照组大鼠明显增加;应用丹红干预8周后,干预组大鼠胆固醇、甘油三酯、低密度脂蛋白胆固醇、血肌酐、尿素氮、24h尿白蛋白定量、肾脏肥大指数、血清vWF水平较糖尿病组明显减低（P＜0.05）,肾组织病理损伤减轻,肾脏vWF表达较糖尿病组明显降低.结论 糖尿病大鼠血清vWF水平及肾组织vWF表达增加,提示vWF与糖尿病肾病关系密切,可能参与了糖尿病肾病的发生、发展;而丹红能通过调节血脂、降低尿白蛋白、降低vWF的表达来发挥肾脏保护作用.     

血小板反应蛋白-1在糖尿病大鼠模型肾损害中的作用及机制初步探讨

目的：观察血小板反应蛋白-1（thrombospondin-1,TSP-1）在链脲佐菌素诱导的糖尿病（diabetes mellitus, DM）大鼠模型肾脏中的表达,初步探讨TSP-1在糖尿病肾病大鼠发病机制中的作用及与糖尿病肾损害的关系。方法：成年雄性SD大鼠80只,随机分为正常对照组（ n＝15）和实验组（ n＝65）。采用STZ 70 mg/kg单次腹腔注射建立DM大鼠模型,按照成模标准去除未成模大鼠。检测各组大鼠空腹血糖（ fasting blood glucose,FBG）、尿蛋白及肾功能;分别于2,4,8周处死各组大鼠,通过HE染色观察各组大鼠肾脏的病理改变,免疫荧光观察TSP-1在各组大鼠肾脏的分布;PCR检测肾组织TSP-1和转录生长因子β1（ transforming growth factor,TGF-β1）基因表达水平;采用酶联免疫吸附法（ ELISA）检测肾组织活性TGF-β1水平;Western杂交检测TSP-1的动态变化。分别对尿蛋白、肾功能、肾脏病理损害、活性TGF-β1与TSP-1蛋白表达的相关性进行分析。结果：HE染色可见DM组大鼠肾小球系膜基质增生,肾小球可见分叶现象、K-W结节等典型的糖尿病肾病损伤特点。与正常对照组相比,DM组大鼠的FBG、尿蛋白、肌酐、尿素氮水平均显著增加（ P﹤0．05）,TSP-1在DM组大鼠主要分布于肾小管、肾小球系膜细胞,其蛋白表达在2周时即明显增加,一直持续到第8周。DM 组大鼠肾脏TSP-1的积分光密度（ integrated optical density,IOD）与尿蛋白、肾脏病理损害、肌酐、尿素氮水平、活性TGF-β1浓度均呈正相关。结论：持续高血糖状态可以诱导大鼠肾脏病理损害,引起尿蛋白升高、肾功能异常;高血糖可诱导大鼠肾脏TSP-1蛋白高表达。TSP-1蛋白的高表达可能通过激活TGF-β1信号通路参与DM大鼠蛋白尿的产生及肾损害的发生发展。     

尾加压素Ⅱ在糖尿病肾病大鼠动脉粥样硬化形成的表达及黄芪的干预

目的:观察糖尿病肾病大鼠主动脉、肾脏组织中尾加压素Ⅱ(urotensinⅡ,UⅡ)及其G蛋白耦联受体14(G-protein-coupled receptor14,GPR14)水平的变化,探讨UⅡ在糖尿病肾病动脉硬化形成的作用和发病机制及黄芪的干预效果。方法:采用高糖高脂饮食和链脲佐菌素(STZ)腹腔注射法建立大鼠糖尿病肾病模型。将大鼠分为正常对照组(NC)、糖尿病肾病组(DN)、黄芪治疗组(HZ)(予黄芪注射液5ml/kg灌胃)。于14周末处死动物,常规病理学检查,免疫组化观察UⅡ、GPR14蛋白表达;RT-PCR观察UⅡmRNA表达。结果:与NC组比较,DN组主动脉、肾组织UⅡ、GPR14表达增加,肾组织UⅡmRNA表达增加(P<0.01);与DN组比较,HZ组表达显著减少(P<0.05)。结论:UⅡ及其受体GPR14的高表达提示其在糖尿病肾病动脉粥样硬化形成的过程中可能起重要作用;黄芪能够缓解肾脏病理损伤,其机制可能是通过抑制UⅡ和GPR14异常表达有关。     

血管内皮生长因子和内抑素在STZ大鼠肾脏中的表达及意义

目的：研究血管内皮生长因子（VEGF）和内抑素（ENS）在2型糖尿病大鼠肾脏中的表达状况、比值变化及与肾脏微血管病变的关系。方法：将Wister大鼠设正常对照组和模型组，使用STZ诱导形成2型糖尿大鼠肾病模型，分别于2、4、8、12周，采用RT-PCR的方法观察大鼠肾脏VEGF和ENSmRNA的表达，并观察其比值变化，同时采用免疫组化观察VEGF蛋白的表达变化。结果：（1）糖尿病组2周时肾脏中VEGFmRNA开始上调（P〈0．05），4、8、12周时较正常对照组明显上调（P〈0．01）。（2）糖尿病组2周时肾脏中ENSmRNA的表达开始上调（P〈0，05），8、12周时表达明显上调（P〈0．01）。（3）糖尿病组2周时肾脏的VEGFmRNA／ENSmRNA值未见变化（P〉0．05），4周时升高（P〈0．05），12周时明显升高（P〈0．01）。（4）免疫组化显示糖尿病肾病组2周时VEGF升高，12周时升高更明显。结论：VEGF和ENS同时参与了糖尿病肾病的血管生成调控，两者表达水平的失衡是其新生血管形成的关键。     

尾加压素Ⅱ与糖尿病肾病大鼠动脉粥样硬化形成的相关性研究及川穹嗪的干预作用

目的:本实验利用链脲佐菌素建立糖尿病肾病大鼠模型,采用免疫组织化学等方法,探讨血浆尾加压素Ⅱ与糖尿病肾病大鼠动脉粥样硬化形成的相关性及其川穹嗪对二者的影响。方法:采用高糖高脂饮食合并链脲佐菌素腹腔注射的方法建立糖尿病肾病大鼠模型。将大鼠随机分为正常对照NC组、糖尿病肾病DN组、川穹嗪CZ组(150mg·kg-1·d-1灌胃)(人与大鼠剂量之比约为7∶1,人常规剂量80～160mg/d除以标准体重×7,所得约为150～300mg/kg;用药方式选为灌胃,应静脉临床试验中难以进行),检测各组大鼠重量、24h尿白蛋白量、血糖、血脂、肾功能等;行HE染色观察各组大鼠病理学形态;免疫组化观察UⅡ、GPR14蛋白表达;RT-PCR观察UⅡmRNA表达。结果:与NC组相比,DN组各项检查及病理学改变明显增大,免疫组化中UⅡ、GPR14蛋白表达增加;RT-PCR观察UⅡmRNA表达增加,差异有统计学意义(P<0.05);而CZ组相对DN组各项结果有所改善(P<0.05)。结论:肾小球UⅡ及其受体GPR14的高表达,提示其在糖尿病肾病动脉粥样硬化形成过程中可能起重要作用;川穹嗪可能是通过抑制UⅡ和GPR14异常表达,一定程度上缓解了肾脏病理损伤,保护肾功能。     

益肾胶囊对糖尿病肾病大鼠肾组织JAK/STAT信号通路的影响

目的：观察益肾胶囊对糖尿病肾病（DN）大鼠肾组织JAK/STAT信号通路影响,探讨益肾胶囊对DN大鼠肾脏保护作用的可能机制。方法：将Wistar大鼠制备成DN模型。随机分为4组,即正常对照组（N组）、DN模型组（DN组）、益肾胶囊治疗组（625mg·kg-1.d-1）、氯沙坦钾治疗组（30mg·kg-1.d-1）。实验周期12周。期间检测大鼠血糖和24h尿蛋白定量,通过光镜及电镜观察肾脏组织病理形态学的变化;采用免疫组化方法检测肾组织磷酸化JAK2（p-JAK2）、磷酸化STAT3（p-STAT3）表达及转化生长因子β1（TGF-β1）表达变化。结果：12周末,DN组大鼠肾组织中p-JAK2、p-STAT3、TGF-β1表达显著高于同期正常对照组（P〈0.05）。益肾胶囊治疗组和氯沙坦钾治疗组肾组织中p-JAK2、p-STAT3、TGF-β1表达显著低于同期DN组（P〈0.05）;24h尿蛋白定量显著低于同期DN组（P〈0.05）;病理损伤较同期DN组改善。结论：益肾胶囊可能部分通过抑制DN大鼠肾组织JAK/STAT通路调节肾组织TGF-β1表达,发挥对DN大鼠肾脏的保护作用。     

Effects of bone morphogenetic protein 2 signal pathway on renal artery calcification in progression of diabetic nephropathy

Objective To explore the effects of renal artery calcification on the progression of diabetic nephropathy (DN), the activation and its role of bone morphogenetic protein 2(BMP2) signal pathway in renal artery of rats. Methods Sixty male SD rats were randomly divided into control group(CON group), DN group and DN with vascular calcification group (DN+VDN group). Rats of group DN and DN+VDN were fed with high sugar and fat diet and injected with streptozocin (STZ) into abdominal cavity to induce diabetes. After diabetic models were successfully made, rats of group DN+VDN were treated by vitamin D3 plus nicotine. The rats were sacrificed at 8th, 12th and 16th week respectively and the levels of renal function, blood glucose and 24 h urinary protein (24-h Upro) were measured. The pathologic changes to the renal artery were observed by von-Kossa staining and the calcium content was detected by calcium assay kit. The pathologic changes to the kidney were observed by HE. Immunohistochemistry was applied to detect the protein expression of BMP2/Smad1/Runx2/Osterix signal pathway in the renal artery and real-time PCR were applied to detect the mRNA expression levels of BMP2 and Runx2. Results The calcium content and the deposition of black granules in DN group were significantly higher than those in group CON and lower than DN+VDN group at each time point (P＜0.05). The renal function indices in group DN and group DN+VDN were gradually increased in 8th,12th and 16th weeks, and were higher than those in group CON (P＜0.05). Compared with that in DN group, although the level of BUN, Scr, Cys C and 24-h Upro in DN+VDN group rats were higher at different time point, the level of Cys C at each time point and the level of 24-h Upro in the 16th week showed significant differences (P＜0.05). The pathological damages of the kidney in group DN and DN+VDN showed a continual worsening trend and the pathological changes of the kidney in group DN+VDN were more serious than those in group DN. Furthermore, the levels of BMP2/Smad1/Runx2/Osterix signal protein and BMP2, Runx2 mRNA in DN rats were higher than those in CON group, lower than DN+VDN group at each time point (P＜0.05). Correlation analysis demonstrated that calcium content was positively correlated with serum BUN, Scr, Cys C, 24-h Upro and the expression of BMP2, Runx2 mRNA (r=0.835, 0.705, 0.829, 0.897, 0.641, 0.683, P＜0.01, respectively). Conclusion Renal artery calcification may participate in and promote the progression of DN, and the BMP2 signal pathway may be an important regulating factor in DN with renal artery calcification.     

白细胞介素-17在肾组织的表达与糖尿病肾病关系探讨

目的：观察白细胞介素-17（IL-17）在糖尿病大鼠不同时期血清、尿液中水平及肾脏表达,探讨其与糖尿病肾病发病相关性。方法：64只SD大鼠随机分为对照组及糖尿病组,以链脲佐菌素（50mg/kg）腹腔注射制备糖尿病大鼠模型。于8周、12周末采用ELISA方法检测血清、尿液中IL-17水平;测尿白蛋白排泄率（UAER）、血糖、血肌酐（Scr）、C反应蛋白（CRP）;免疫组织化学染色观察肾组织中IL-17、转化生长因子-β1（TGF-β1）表达。结果：与对照组比较,糖尿病组血清、尿液中IL-17水平8周开始升高,肾组织中IL-17、TGF-β1表达增加,12周升高更明显;相关分析显示肾组织中IL-17表达与TGF-β1表达及尿液中IL-17水平、UAER、血CRP均呈正相关。结论：IL-17在糖尿病大鼠肾小管间质中表达随病程延长而增加,可能参与糖尿病肾病肾小管间质病变的发生发展。     

内皮素受体阻断剂对糖尿病大鼠肾脏血管紧张素Ⅱ1型受体表达的影响

目的：观察糖尿病大鼠肾脏血管紧张素Ⅱ1型（AT1）受体的改变以及内皮素受体阻断剂bosentan对其影响。方法：将SD大鼠建成链脲佐菌素诱导的糖尿病模型,设非治疗组、bosentan治疗组和正常对照组。4周后采用免疫组织化学、Western blot及RT－PCR方法检测肾脏AT1受体基因和蛋白表达。结果：与SD对照组相比,糖尿病大鼠存在明显的蛋白尿和内生肌酐清除率升高,其肾脏AngⅡ水平明显升高,同时伴有AT1受体的mRNA和蛋白表达显著下降。bosentan能显著缓解上述异常。结论：糖尿病大鼠肾脏AngⅡ及AT1受体表达明显异常,bosentan具有治疗作用。     

Sevelamer hydrochloride attenuates kidney and cardiovascular calcifications in long-term experimental uremia

BACKGROUND: In chronic renal failure (CRF), hyperphosphatemia and an elevated calcium-phosphate product are associated with vascular calcification and increased cardiovascular morbidity and mortality. Previous data have demonstrated that 3-month treatment of uremic rats with sevelamer was associated with less nephrocalcinosis compared to calcium carbonate (CaCO3), despite similar control of serum phosphorus, calcium-phosphorus product (Ca x P product), and secondary hyperparathyroidism. There was no evidence of aortic calcification after 3 months of uremia (J Am Soc Nephrol 13:2299-2308, 2002). The present studies explore the influence of sevelamer and CaCO3 on cardiovascular and kidney calcifications in long-term experimental uremia over 6 months. METHODS: Normal and 5/6 nephrectomized rats (U) were fed a high phosphorus (HP) diet for 6 months. Two phosphate binders, CaCO3 and sevelamer, were administered and their influence on hyperphosphatemia, secondary hyperparathyroidism, kidney/myocardial/aortic calcification, and renal function was compared. RESULTS: All uremic rats began the study with the same degree of renal failure. Sevelamer was as effective as CaCO3 in reducing serum phosphorus, Ca x P product, and attenuating secondary hyperparathyroidism. Despite similar serum cholesterol levels, rats in the U-HP + sevelamer group had markedly lower calcium deposition in the myocardium and aorta (myocardium, 72 +/- 4 microg/g wet tissue; aorta, 736 +/- 156 microg/g wet tissue) compared to rats in either the U-HP + CaCO3 group (myocardium, 179 +/- 48, P < 0.05; aorta, 1308 +/- 343, P < 0.05) or the U-HP group (myocardium, 98 +/- 10, NS; aorta, 2150 +/- 447, P < 0.05). Dual immunohistochemical analysis for calcium and endothelial cell markers demonstrated that myocardial calcium deposition was intravascular within capillaries. Furthermore, calcium deposition in the kidney of uremic rats treated with sevelamer (582 +/- 111 microg/g wet tissue) was lower than that found in uremic rats treated with CaCO3 (1196 +/- 180 microg/g wet tissue). Sevelamer-treated rats had less deterioration in renal function with an associated lower serum creatinine, higher creatinine clearance, and less proteinuria. There was no difference in overall mortality between the three experimental groups. CONCLUSION: In long-term experimental CRF, in addition to controlling serum phosphorus and secondary hyperparathyroidism as efficiently as CaCO3, treatment with the phosphate-binder sevelamer attenuates vascular and kidney calcification.     

Aminoguanidine ameliorates overexpression of prosclerotic growth factors and collagen deposition in experimental diabetic nephropathy

Profibrotic cytokines and the formation of advanced-glycation end products (AGE) have both been implicated in the pathogenesis of glomerulosclerosis in diabetic kidney disease. However, tubulointerstitial pathology is also an important determinant of progressive renal dysfunction in diabetic nephropathy. This study sought to investigate the expression of profibrotic growth factors and matrix deposition in the glomerulus and the tubulointerstitium and to examine the effect of blocking AGE formation in experimental diabetic nephropathy. Thirty-six male Sprague-Dawley rats were randomized into control and diabetic groups. Diabetes was induced in 24 rats by streptozotocin. Twelve diabetic rats were further randomized to receive the inhibitor of AGE formation, aminoguanidine (1 g/l drinking water). At 6 mo, experimental diabetes was associated with a three-fold increase in expression of transforming growth factor (TGF)-beta1 (P < 0.01 versus control) and five-fold increase in platelet-derived growth factor (PDGF)-B gene expression (P < 0.01 versus control) in the tubulointerstitium. In situ hybridization demonstrated a diffuse increase in both TGF-beta1 and PDGF-B mRNA in renal tubules. Aminoguanidine attenuated not only the overexpression of TGF-beta1 and PDGF-B but also reduced type IV collagen deposition in diabetic rats (P < 0.05). TGF-beta1 and PDGF mRNA within glomeruli were also similarly increased with diabetes and attenuated with aminoguanidine. The observed beneficial effects of aminoguanidine on the tubulointerstitium in experimental diabetes suggest that AGE-mediated expression of profibrotic cytokines may contribute to tubulointerstitial injury and the pathogenesis of diabetic nephropathy.     

Macrophages in mouse type 2 diabetic nephropathy: correlation with diabetic state and progressive renal injury

BACKGROUND: Macrophage-mediated renal injury has been implicated in progressive forms of glomerulonephritis; however, a role for macrophages in type 2 diabetic nephropathy, the major cause of end-stage renal failure, has not been established. Therefore, we examined whether macrophages may promote the progression of type 2 diabetic nephropathy in db/db mice. METHODS: The incidence of renal injury was examined in db/db mice with varying blood sugar and lipid levels at 8 months of age. The association of renal injury with the accumulation of kidney macrophages was analyzed in normal db/+ and diabetic db/db mice at 2, 4, 6, and 8 months of age. RESULTS: In db/db mice, albuminuria and increased plasma creatinine correlated with elevated blood glucose and hemoglobin A1c (HbA1c) levels but not with obesity or hyperlipidemia. Progressive diabetic nephropathy in db/db mice was associated with increased kidney macrophages. Macrophage accumulation and macrophage activation in db/db mice correlated with hyperglycemia, HbA1c levels, albuminuria, elevated plasma creatinine, glomerular and tubular damage, renal fibrosis, and kidney expression of macrophage chemokines [monocyte chemoattractant protein-1 (MCP-1), osteopontin, migration inhibitory factor (MIF), monocyte-colony-stimulating factor (M-CSF)]. The accrual and activation of glomerular macrophages also correlated with increased glomerular IgG and C3 deposition, which was itself dependent on hyperglycemia. CONCLUSION: Kidney macrophage accumulation is associated with the progression of type 2 diabetic nephropathy in db/db mice. Macrophage accumulation and activation in diabetic db/db kidneys is associated with prolonged hyperglycemia, glomerular immune complex deposition, and increased kidney chemokine production, and raises the possibility of specific therapies for targeting macrophage-mediated injury in diabetic nephropathy.     

益肾胶囊对糖尿病肾病大鼠肾小管间质sFRP-1表达的影响

目的:观察益肾胶囊对糖尿病肾病大鼠肾小管间质Wnt通路抑制因子-分泌型卷曲相关蛋白-1(sFRP-1)表达的影响。方法:采用一次性腹腔注射链脲佐菌素(STZ)法诱导建立糖尿病肾病(DN)模型,实验分组为:正常对照组(对照组)、DN模型对照组(模型组)、益肾胶囊组、氯沙坦组,每组8只。益肾胶囊组每只大鼠灌胃益肾胶囊625 mg.kg-1.d-1,氯沙坦组每只灌胃氯沙坦20 mg.kg-1.d-1,正常组和模型组每日给予等量生理盐水灌胃,于2、4、8、12周末测定24 h尿蛋白定量,于治疗12周后测定各组血糖、血尿素氮(BUN)、肌酐(Scr),同时观察肾小管病理损伤,并采用免疫组化和实时荧光定量PCR方法观察大鼠肾小管间质中sFRP-1、Wnt通路关键调节因子β-catenin表达情况及肾组织中sFRP-1、β-catenin mRNA的表达。结果:12周末,与对照组相比,模型组大鼠血糖、24 h尿蛋白定量、肾小管损伤指数均显著上升,免疫组化示:肾小管间质中sFRP-1表达增加,β-catenin出现胞浆或(和)核表达,实时荧光定量PCR结果显示二者mRNA表达上调(P<0.05);益肾胶囊治疗后,与模型组相比,肾小管间质中sFRP-1表达进一步增加,肾组织中sFRP-1 mRNA进一步上调,β-catenin胞浆或(和)核表达减少,β-catenin mRNA表达下调(P<0.05)。结论:益肾胶囊可能通过上调sFRP-1在DN大鼠肾小管间质的表达,发挥对DN大鼠肾脏的保护作用。