The effect of irbesartan on the expression of angiopoietin-like protein 2 in the kidneys of type2 diabetes rats

Objective To observe the effect of irbesartan on the expression of angiopoietin-like protein 2 (ANGPTL2) in the diabetic rats kidney and explore the underlying mechanism. Methods A total of sixty male SD rats were divided into normal control group (NC group, n=15) and experimental group (n=45) randomly. The experimental group was fed with high sugar-fat diet and given a low dose streptozocin（STZ 30 mg/kg）to establish type 2 diabetic model. Rats successfully induced diabetes were randomly divided into 2 groups: diabetes group (DM) and irbesartan group (DI). Weight, blood pressure, blood glucose, serum creatinine (Scr), blood urea nitrogen(BUN), 24 hour urinary albumin(UAL) and renal histomorphology were observed after drug intervention at the 4th, 8th and 12th weeks. The expression of ANGPTL2 in renal tissue were detected by immunohistochemistry, real-time PCR and Western blotting. Results The levels of Scr, BUN, TG, TC and UAL in group DM were higher than in group NC at the 4th, 8th and 12th week (all P＜0.05).Compared with that in group DM, above indexes were lower in group DI at the 4th, 8th and 12th week (all P＜0.05). The pathological changes of the kidney in group DM were more serious than that in group DI. The expression of ANGPTL2 in group DM was much higher than that in group NC at the 4th, 8th and 12th week (all P＜0.05), and irbesartan treatment inhibited the up-regulation of ANGPTL2 in group DI(all P＜0.05). Conclusion The expression of ANGPTL2 increases in T2DM rats kidney tissue with time and irbesartan can inhibit the up-regulation of ANGPTL2 in T2DM rats.     

洛沙坦对糖尿病大鼠肾脏炎症反应及足细胞损伤的影响

目的 探讨洛沙坦对糖尿病大鼠模型肾组织炎症反应及足细胞损伤的影响。方法 Wistar大鼠随机分为3组：对照组（NC）、糖尿病组（DM）、洛沙坦治疗组（DL）。链脲菌素（STZ）注射建立糖尿病动物模型，洛沙坦治疗组在大鼠建立糖尿病同时即开始以洛沙坦（20mg·kg·d^-1）灌胃治疗。分别于2周和12周杀鼠，观察体重、肾重、肾重／体重指数、尿蛋白量（24h）、Scr、Ccr以及肾组织病理改变；免疫组化染色观察肾小球细胞间黏附分子（ICAM-1）、白细胞共同抗原（CD45）、nephrin、血管内皮生长因子（VEGF）表达的改变；免疫印迹观察肾组织nephrin、VEGF表达的改变。结果 （1）DL组大鼠肾重／体重指数、尿蛋白量（24h）、Ccr则均低于DM组，差异均有统计学意义（P均〈0．05），其病理改变程度也比DM组轻。（2）免疫组化及免疫印迹显示，DM组和DL组肾小球ICAM-1、CD45、VEGF表达均高于对照组，而DL组ICAM-1、CD45、VEGF表达均低于DM组；DM组和DL组大鼠肾小球nephrin表达低于对照组；DL组肾小球nephrin表达高于DM组，差异均有统计学意义（P均〈0．01）。结论 洛沙坦可能通过抑制肾组织白细胞浸润、炎症反应及减轻足细胞损伤，达到肾脏保护作用。     

MCP-1、ICAM-1在糖尿病肾病大鼠肾脏损害中作用及厄贝沙坦干预的影响

目的：探讨MCP-1、ICAM-1在糖尿病肾病大鼠肾脏损害中作用及厄贝沙坦对二者的影响.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、厄贝沙坦DI组,检测各组大鼠24 h尿量、24 h尿白蛋白定量（24 h UTP）、血糖（BG）、血肌酐（Scr）、尿素氮（BUN）、肾重指数（KI）;行HE染色观察各组大鼠病理学形态,免疫组化观察MCP-1、ICAM-1蛋白的表达,RT-PCR观察MCP-1 mRNA、ICAM-1mRNA的表达.结果：与NC组比较,DN组大鼠肾脏病理改变加重,24 h尿量、24 h UTP、KI、BG、Scr、BUN、肾脏组织中MCP-1mRNA和ICAM-1mRNA及蛋白水平均显著增加,差异均有统计学意义（P＆lt;0.01）;与DN组比较,DI组大鼠BG、BUN、Scr有所改善,差异无统计学意义;肾脏病理改变减轻,其余指标明显降低,差异有统计学意义（P＆lt;0.05）.结论：MCP-1、ICAM-1在糖尿病肾病肾脏损害过程中可能起重要作用;厄贝沙坦能够减轻糖尿病肾病肾组织MCP-1、ICAM-1的表达,缓解了肾脏病理损伤.     

霉酚酸酯及缬沙坦对糖尿病大鼠足细胞的保护机制研究

目的探讨霉酚酸酯、缬沙坦及2者联合应用对糖尿病。肾病（DN）大鼠足细胞损伤的保护作用。方法雄性Wistar大鼠行右肾切除后，腹腔注射链脲佐菌素（STZ，65mg／kg）建立糖尿病模型。将实验动物随机分为右。肾切除对照组（NC）、糖尿病组（DM）、霉酚酸酯治疗组（M）、缬沙坦治疗组（V）、缬沙坦和霉酚酸酯联合治疗组（V＋M）。治疗组分别给予霉酚酸酯15mg·kg^-1·d^-1，缬沙坦40mg·kg^-1·d^-1；联合治疗组为上述两组之和。检测各组8周末的左肾质量／体质量比值、尿蛋白量（24h）、血糖（Glu）、Scr。光镜及电镜观察肾组织形态学变化。免疫组化检测肾组织中nephrin、结蛋白（desmin）及单核细胞趋化因子1（MCP-1）蛋白表达。实时PCR测定肾组织中nephrin及MCP-1mRNA表达。结果与NC组相比，DM组大鼠血糖、尿蛋白量及左肾质量／体质量比值均显著上升（P〈0．01）；肾小球硬化指数（GSI）及肾间质损害加重（P〈0．01）；肾组织内MCP-1、desmin蛋白表达均显著上调（P〈0．01）。与DM组比较，M组、V组及V＋M组上述指标除Glu、Scr外，均明显改善（P〈0．05或P〈0．01）。与NC组（100％）相比，DM组nephrinmRNA表达下调（78％，P〈0.05）；各治疗组nephrinmRNA表达增加，以M组增加最明显（134％，P〈0．01）。与NC组（100％）相比，DM组MCP-1mRNA表达明显上调（251％，P〈0.05）；各治疗组明显降低，以M组最显著（126％，P〈0．01）。nephrinmRNA与MCP-1mRNA表达呈负相关（r=-0，86。P〈0．01）。尿蛋白量（24h）与MCP-1mRNA呈正相关fr=0．82，P〈0．01）；与nephrinmRNA呈负相关（r=-0．78，P〈0．01）。结论霉酚酸酯及缬沙坦均能下调糖尿病大鼠肾组织中desmin及MCP-1基因及蛋白的表达，上调nephrin基因及蛋白表达，降低尿蛋白量，预防肾损伤。联合治疗不优于单一治疗。霉酚酸酯可能通过抗炎性反应减轻足细胞损伤，减少蛋白尿，对早期DN大鼠具有明显的肾保护作用。     

舒洛地特对糖尿病大鼠肾脏组织VEGF和VEGF-R2蛋白表达的影响

目的:观察舒洛地特对糖尿病大鼠肾脏的保护作用并探讨其机制。方法:采用链脲佐霉素(STZ,60mg/kg)腹腔注射法构建1型糖尿病大鼠模型,随机分为4组:糖尿病组(DM组)、舒洛地特10mg·kg-1·d-1组(S10组)、舒洛地特20mg·kg-1·d-1组(S20组)、氯沙坦30mg·kg-1·d-1组(L组),每组各10只。另取10只未造模大鼠作正常对照组(N组)。干预12周后采集各组大鼠血、尿标本和肾脏组织标本,观察和分析各组大鼠肾脏功能和结构的改变;Westernblot-ting和免疫组织化学法检测肾组织VEGF及其受体VEGF-R2的表达。结果:同N组相比,DM组大鼠24h尿白蛋白定量、血肌酐及尿素氮显著增加(P<0.01),病理改变较明显。同DM组相比,治疗组(S10、S20及L组)大鼠24h尿白蛋白定量减少(P<0.05或P<0.01);同S10组相比,S20及L组大鼠24h尿白蛋白定量减少(P<0.05)。光镜及电镜显示治疗组较DM组病理变化减轻,尤以S20及L组病变减轻明显。大鼠肾脏组织VEGF和VEGF-R2的表达,DM组明显高于N组(P<0.01),治疗组显著低于DM组(P<0.05或P<0.01)。结论:舒洛地特对糖尿病大鼠肾脏有保护作用,而20mg·kg-1·d-1较10mg·kg-1·d-1的剂量效果更好。下调肾脏组织VEGF和VEGF-R2蛋白表达可能是舒洛地特对糖尿病大鼠肾脏保护作用机制之一。     

舒洛地特对糖尿病大鼠肾脏病变和SOCS-1、TGF-β1表达的影响

目的：观察舒洛地特对糖尿病大鼠肾脏的保护作用并探讨其机制.方法：采用STZ（链脲佐霉素,60 mg/kg）腹腔注射法构建1型糖尿病大鼠模型,随机分为4组：糖尿病组（DM组）、舒洛地特10 mg·kg-1·d-1组（S10组）、舒洛地特20 mg·kg-1·d-1组（S20组）、氯沙坦30 mg·kg-1·d-1组（L组）,每组各10只.另取10只未造模大鼠作正常对照组（N组）.干预12周后采集各组大鼠血、尿标本和肾脏组织标本,观察和分析各组大鼠肾脏功能和结构的改变;Western blotting和免疫组织化学法检测肾组织SOCS-1及TGF-β1蛋白的表达.结果：同N组相比,DM组大鼠24 h尿白蛋白定量、血肌酐及尿素氮显著增加（P〈0.01）,病理改变较明显.同DM组相比,治疗组（S10、S20及L组）大鼠24 h尿白蛋白定量减少（P〈0.05或P〈0.01）;同S10组相比,S20及L组大鼠24 h尿白蛋白定量减少（P〈0.05）.光镜及电镜显示治疗组较DM组病理变化减轻,尤以S20及L组病变减轻明显.大鼠肾脏组织SOCS-1的表达,DM组明显高于N组（P〈0.05）,治疗组显著高于DM组（P〈0.05或P〈0.01）.大鼠肾脏组织TGF-β1的表达,DM组明显高于N组（P〈0.01）,治疗组显著低于DM组（P〈0.05或P〈0.01）.结论：舒洛地特对糖尿病大鼠肾脏有保护作用,而20 mg·kg-1·d-1较10 mg·kg-1·d-1的剂量效果更好.上调肾脏组织SOCS-1的表达,抑制TGF-β1的过表达可能是舒洛地特对糖尿病大鼠肾脏保护作用机制之一.     

血管紧张素1-7对糖尿病大鼠肾小管间质纤维化的影响

目的 研究血管紧张素1-7(Ang1-7)对糖尿病大鼠肾小管间质纤维化的影响及其可能机制.方法 32只雄性Wistar大鼠被随机分为4组:健康对照组(NC组)、模型组(DM组)、替米沙坦组(TM组)、治疗组(T组).建模成功后第9周末检测各组大鼠24 h尿蛋白量、尿NAG/Cr、血糖、血胰岛素、三酰甘油(TG)、总胆固醇(TC)、BUN、Scr、血K+及血Na+ ;PAS染色观察肾脏病理改变 ;实时定量PCR法检测各组大鼠肾脏组织中转化生长因子β1(TGF-β1)、过氧化物酶体增殖物激活受体(PPAR)γ、α平滑肌肌动蛋白(α-SMA)mRNA水平 ;Western印迹法检测PPARγ、α-SMA、TGF-β1蛋白表达.结果 (1)第9周末,DM组大鼠血压、尿蛋白量、肾质量/体质量较NC组显著升高(P＜0.05),TM组及T组较DM组显著降低(P＜0.05),且T组变化更明显.(2)DM组第9周末肾间质损伤指数显著高于NC组(P＜0.05),TM组及T组则低于DM组(P＜0.05).(3)实时定量PCR结果显示,DM组TGF-β1、α-SMAmRNA水平显著升高(P＜0.05),PPARγ mRNA水平显著下降(P＜0.05),TM组及T组较DM组TGF-β1、α-SMA mRNA水平均显著下降(P＜0.05),PPARγ mRNA水平显著上升(P＜0.05),且T组变化更明显.(4)Western印迹结果显示,DM组TGF-β1、α-SMA蛋白水平显著升高(P＜0.05),PPARγ蛋白水平显著下降(P＜0.05),TM组及T组较DM组TGF-β1、α-SMA蛋白水平均显著下降(P＜0.05),PPARγ蛋白水平显著上升(P＜0.05),且T组变化更明显.结论 Ang1-7在体内可通过上调PPARγ表达,抑制α-SMA表达,对糖尿病大鼠肾小管间质纤维化可能具有抑制作用.     

舒洛地特对糖尿病大鼠肾脏病变的保护作用与机制探讨

目的 观察舒洛地特对糖尿病大鼠肾脏组织核因子NF-κB活性及单核细胞趋化蛋白1(MCP-1)表达的影响,探讨舒洛地特对糖尿病肾病的保护机制.方法 高脂高糖喂养联合小剂量链脲佐菌素腹腔注射建立Wistar大鼠糖尿病模型,并按随机数字表法分为非治疗组(DM)及舒洛地特治疗组( DMS).非糖尿病大鼠作为正常对照组(NC).12周后杀检,测定血糖、血肌酐、尿素氮、三酰甘油、胆固醇;免疫比浊法测定24h尿白蛋白;光镜下观察肾小球形态和结构,计算平均肾小球体积;免疫组化法检测肾组织MCP-1表达;Western印迹方法测定NF-κB活性.结果 与NC组比较,DM组及DMS组血糖、三酰甘油、胆固醇均显著升高(均P＜ 0.01);DMS组与DM组比较,以上指标差异无统计学意义.与NC组相比,DM组及DMS组血肌酐、尿素氮、24 h尿白蛋白显著升高(均P＜0.01).与DM组比较,DMS组血肌酐[(39.1±0.88) μmol/L比(41.0±2.16) μmol/L,P＜0.05]、尿素氮[(9.12±1.06) mmol/L比(9.87±0.19) mmol/L,P＜0.05]、24h尿白蛋白[(19.92±0.96) mg/24 h比(25.99±0.52) mg/24 h,P＜ 0.01]均显著降低.与NC组比较,DM组平均肾小球体积显著增加[(7.47±1.11)×105 μm3比(4.22±1.09)×105μm3,P＜ 0.01];DMS组平均肾小球体积[(6.64±0.71 )×105 μm3,P＜0.05]较DM显著降低,但仍显著高于对照组(P＜0.01).与NC组相比,DM组肾组织MCP-1表达显著升高[( 12.17±1.94 )/HPF比(1.19±0.70)/HPF,P＜0.01];与DM组比较,DMS组肾组织MCP-1表达[(9.22±1.61 )/HPF,P＜0.01]显著降低,但仍高于NC组(P＜0.01).与NC组相比,DM组肾组织NF-κB活性显著升高[(0.89±0.07)比(0.24±0.03),P＜0.01];与DM比较,DMS组肾组织NF-κB活性[(0.27±0.01),P＜0.01]显著降低,与NC组比较,差异无统计学意义.结论 舒洛地特对糖尿病肾病具有防治作用,抑制NF-κB活性及MCP-1表达可能是其作用机制之一.     

肿瘤坏死因子相关的凋亡诱导配体系统和核因子κB在糖尿病大鼠肾脏中的表达

目的探讨肿瘤坏死因子相关的凋亡诱导配体（TRAIL）系统在糖尿病肾病发生发展中的作用。方法观察TRAIL及其死亡受体4（DR4）、诱骗受体2（DcR2）和核因子KB（NF-KB）在糖尿病大鼠不同时期肾脏中的表达及分析其与肾功能的关系。将80只Wistar大鼠随机分为对照组（NC组）和糖尿病组（DM组），一侧肾切除后，腹腔注射链脲佐菌素（STZ，55mg／kg）建立糖尿病大鼠模型。在第4、8、12、16周末，随机处死各组8只大鼠，收集血液、尿液和肾组织，检测血生化指标、尿蛋白量（24h）和肥大指数等。应用荧光实时定量RT-PCR和免疫组织化学的方法检测肾皮质TRAIL及其受体DR4、DcR2和NF-KB的mRNA和蛋白的表达情况。结果各时间点DM组大鼠尿蛋白量（24h）和肥大指数（肾质量，体质量）均显著高于NC组（P〈0．05）；血白蛋白在第8周末开始显著低于NC组（P〈0．01）；Scr、BUN于第16周末显著高于NC组（P〈0．01）。DM组大鼠TRAIL及其受体DR4mRNA在第4、8及12周末时表达均显著低于NC组（P〈0．01）；第16周末时表达显著高于NC组（P〈0．01）。DM组大鼠DcR2mRNA在第4、8及12周末时表达均显著高于NC组（P〈0．01）；NF-KB的基因表达均显著高于NC组（P〈0．05）。免疫组化结果显示TRAIL及其受体DR4、DcR2主要在肾小管表达，而在肾小球和脉管系统未见表达；NF-KB在肾小球和肾小管均有表达；TRAIL及其受体和NF-KB在各组表达趋势与PCR结果一致。结论TRAIL系统作为调节细胞凋亡的一组重要因子，参与了糖尿病肾病的发生发展。     

舒洛地特对糖尿病大鼠肾脏病变的影响

目的:探讨2种剂量舒洛地特对糖尿病大鼠肾脏病变的影响。方法:采用STZ(链尿佐霉素,60mg/kg)腹腔注射法构建1型糖尿病大鼠模型,随机分为4组:糖尿病组(DM组)、舒洛地特10mg·kg-1·d-1组(S10组)、舒洛地特20mg·kg-1·d-1组(S20组)、氯沙坦30mg·kg-1·d-1组(L组),每组各10只。另取10只未造模大鼠作正常对照组(N组)。干预12周后测体重、24h尿白蛋白定量、血糖、血肌酐、尿素氮及肾重,光镜、电镜观察肾组织形态、结构的变化。结果:同N组相比,DM组大鼠24h尿白蛋白定量、血肌酐及尿素氮显著增加(P<0.01),病理改变较明显。同DM组相比,治疗组(S10、S20及L组)大鼠24h尿白蛋白定量减少(P<0.05或P<0.01),但血肌酐及尿素氮下降差异无统计学意义(P>0.05);同S10组相比,S20及L组大鼠24h尿白蛋白定量减少(P<0.05)。光镜及电镜显示治疗组较DM组病理变化减轻,尤以S20及L组病变减轻明显。结论:舒洛地特可对糖尿病大鼠有肾脏保护作用,而20mg·kg-1·d-1较10mg·kg-1·d-1的剂量效果更好。     

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.     

Adonesine A1 receptor and megalin defect in diabetic mice with early kidney disease

Objective To observe the effect of adenosine A1 receptor (A1AR) on the megalin defect in type 1 diabetic mice with early kidney disease. Methods 7-8 week-old, baseline body weight and fasting blood glucose matched wild type (WT) C57BL/6J mice were selected, and randomly divided into two groups: control group (n=6) and WT DM group (n=6). In the same way, male A1AR knock-out C57BL/6J mice were selected as A1AR-/- DM group (n=6). DM model was established by intraperitoneal injection of streptozocin. The blood glucose (BG), body weight (BW), kidney weight (KW), 24 h proteinuria (24hUP) and albumin creatine ratio (ACR) were measured at 4 weeks. The renal pathological lesion was observed and the expression of megalin in proximal tubules was examined by immunohistochemistry. The expression of caspase-1, IL-18 and A1AR were detected by Western blotting. Results At 4th week, compared with WT control mice, the BG, BW, KW and 24hUP of WT DM mice were increased significantly (n=6, P＜0.01), with the pathological glomerular enlargement, mesangial cell proliferation, extracellular matrix accumulation and renal tubule hypertrophy being observed. Immunohistochemistry revealed decreased expression of megalin, an important multiligand protein receptor on the brush border of proximal tubular epithelial cells in WT DM mice, which was correlated with 24hUP (r=-0.645, P＜0.01). Compared with the control mice, the expressions of caspase-1, IL-18 and A1AR were significantly increased in WT DM mice (P＜0.05). For A1AR-/- DM mice, more serious pathological lesion and megalin defect, together with increasing of casapase-1 and heavier proteinuria were observed than those in WT DM mice. Conclusion A1AR may play a protective role in megalin expression of diabetic mice with early kidney disease, in which the mechanism may be associated with caspase-1 related pyroptosis pathway. The details need further exploration.     

Mycophenolate mofetil prevents the progressive renal failure induced by 5/6 renal ablation in rats

BACKGROUND: Extensive renal ablation is associated with progressive sclerosis of the remnant kidney. Because lymphocytes and monocytes accumulate in the remnant kidney, it is likely that they play a role in the renal scarring. Therefore, we treated rats with 5/6 nephrectomy (5/6Nx) with mycophenolate mofetil (MMF), a drug that has an antiproliferative effect and that suppresses the expression of intercellular adhesion molecules. METHODS: Sprague-Dawley rats with 5/6Nx received MMF (30 mg. kg-1. day-1 by daily gastric gavage, N = 15) or vehicle (N = 16). Ten additional rats were sham operated. All rats were fed a 30% protein diet. Body weight, serum creatinine, and urinary protein excretion were determined weekly. Lipid peroxidation, as a measure of oxidative stress observed by urinary malondialdehyde determinations, was performed every two weeks. Histologic studies were done in the remnant kidney four weeks (9 rats from the vehicle-treated group, 7 rats from the MMF group, and 5 sham-operated rats) and eight weeks after surgery (the remaining rats). Glomerular volume, sclerosis in glomeruli (segmental and global) and interstitium (semiquantitative scale), infiltrating lymphocytes and macrophages (CD43- and ED1-positive cells), and expression of adhesion molecules (CD54, CD18, and CD11b) were analyzed. RESULTS: MMF treatment prevented the progressive increment in serum creatinine and the proteinuria observed in the 5/6 nephrectomized rats during the eight weeks of observation (P < 0.01). Weight gain was comparable in the MMF-treated and sham-operated rats, whereas weight gain was decreased in untreated 5/6 nephrectomized rats. Excretion of malondialdehyde increased after surgery but returned sooner to control levels in the MMF-treated rats. Increments in glomerular size and mean arterial blood pressure induced by renal ablation were not modified by MMF treatment. Eight weeks after surgery, segmental sclerosis was present in 48.4 +/- 8.35% (+/- sd) glomeruli in the vehicle-treated group versus 25 +/- 10.5% in the MMF-treated group (P < 0.001). Interstitial fibrosis was reduced significantly with MMF treatment (P < 0.001). Infiltration with CD43- and ED1-positive cells in glomeruli and interstitium was two to five times lower in MMF-treated rats (P < 0.01). Expression of adhesion molecules CD18 and CD11b was similarly reduced. CONCLUSION: MMF ameliorates the progressive renal damage in the remnant kidney after 5/6Nx. This effect is associated with a reduction in the infiltration of lymphocytes and monocytes, whereas glomerular hypertrophy and systemic hypertension are unchanged.     

他克莫司与霉酚酸酯治疗儿童难治性IgA肾病的疗效比较

目的比较他克莫司(tacrolimus,TAC)与霉酚酸酯(mycophenolate mofetil,MMF)在难治性IgA肾病(IgA nephropathy,IgAN)患儿的临床疗效。方法难治性IgAN的诊断定义为:在联合肾素-血管紧张素系统(RAS)阻断剂及糖皮质激素序贯治疗后,仍表现为大量蛋白尿(≥50 mg·kg-1·d-1)。遵循病例对照匹配方法,回顾性选取2012年1月1日至2016年12月31日在东部战区总医院行肾活检确诊为难治性IgAN的患儿76例,根据治疗方案将患儿分为TAC组(38例)和MMF组(38例),比较两组24 h尿蛋白量(24hUP)、血清白蛋白(Alb)、血清肌酐(Scr)、血清尿酸(UA)、血糖(Glu)、不良反应发生情况以及疗效等。结果两组患者间年龄、性别比、血压、估算肾小球滤过率(eGFR)、24hUP、尿红细胞计数(U-RBC)、Scr、Alb、BUN、天门冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、Glu及病理牛津分型、加用免疫抑制剂前行大剂量甲泼尼龙冲击治疗比例差异均无统计学意义(均P>0.05),两组患者具有可比性。用药3个月起,两组患儿24hUP均较基线值明显降低,差异有统计学意义(均P<0.05),且在3、6、12个月时TAC组24hUP均显著低于MMF组(均P<0.05)。TAC组Alb在用药1个月后即较基线值明显升高(P<0.05),而MMF组在用药3个月后明显升高(P<0.05),其中TAC组Alb在1、3、6个月时均高于MMF组(均P<0.05),在12个月时两组Alb差异无统计学意义。TAC组的总有效率、完全缓解率及无效率自用药3个月起与MMF组差异均有统计学意义(均P<0.05),但随访期间部分缓解率、随访时点复发率及累计复发率两组间差异无统计学意义(均P>0.05)。其中TAC组在6个月时达到最大有效率(94.7%),而MMF组在12个月时达到最大有效率(68.4%),差异有统计学意义(χ2=8.756,P=0.003)。两组不良反应发生率比较差异无统计学意义(15.8%比21.1%,χ2=0.350,P=0.554)。但TAC组在治疗3个月时血糖高于MMF组,差异有统计学意义[5.02(4.72,5.22)mmol/L比4.42(4.19,5.07)mmol/L,Z=-2.745,P=0.006]。结论TAC与MMF治疗难治性IgAN均取得良好的治疗效果,但TAC组更快达到缓解水平,且缓解率更高。     

全反式维甲酸延缓糖尿病肾脏纤维化进展的实验研究

目的：探讨全反式维甲酸对延缓糖尿病大鼠肾脏纤维化的保护作用。方法：随机选择雄性SD大鼠分为空白对照组（NC）与造模组,制作糖尿病模型分为模型组（DM）与维甲酸治疗组（DM＋atRA）。于模型成型后分别于第8、12周观察各组大鼠24h尿蛋白（Ualb）,内生肌酐清除率（Ccr）,肾重／体重。肾脏病理及电镜观察肾小球基底膜厚度,免疫组织化学方法观察肾组织Ⅳ、Ⅲ型胶原表达。结果：维甲酸组肾重／体重、Ualb、Ccr较同时期模型组下降（P＜0．05）,且12周大鼠肾组织细胞外基质（ECM）重要成分ColⅣ表达（5．75±0．41）、ColⅢ表达（4．98±1．37）与模型组比较显著减少（P＜0．01）。病理观察维甲酸组与模型组比较肾小球系膜细胞和内皮细胞增生减轻,足突融合改善,系膜基质增生缓解,基底膜厚度变薄,减轻了肾脏病理损害。结论：全反式维甲酸能减轻糖尿病大鼠早期肾脏病理损伤,减少细胞外胶原基质积聚,延缓肾脏纤维化。     

乙酰肝素酶在糖尿病肾病大鼠蛋白尿发生中的作用

目的 观察糖尿病肾病大鼠肾组织乙酰肝素酶（HPA）的表达，探讨其在糖尿病肾病大鼠蛋白尿发生中的作用。 方法 SD健康大鼠被随机分为健康对照组（n = 6）、糖尿病6周组(DM6w, n = 10)和糖尿病12周组(DM12w, n = 10)，采用一次性腹腔注射链脲菌素（STZ）的方法建立糖尿病大鼠模型。分别于造模后6周和12周末测定各组大鼠相对肾质量、血糖、尿素氮、血肌酐、24 h尿量及尿蛋白量(24 h)等指标，并观察肾脏病理改变。RT-PCR和免疫组织化学法检测各组大鼠肾组织HPA mRNA和蛋白表达变化，并分析其与蛋白尿发生的相关性。 结果 （1）DM6w和DM12w组大鼠的相对肾质量、血糖、尿素氮、24 h尿量及尿蛋白量(24 h)与健康对照组相比明显升高, 差异均有统计学意义(P < 0.05或P < 0.01)。（2）DM6w和DM12w组大鼠HPA mRNA和蛋白表达比健康对照组均显著增高(P < 0.01)。（3）大鼠肾组织HPA mRNA和蛋白表达与尿蛋白量(24 h)之间均呈正相关 (r = 0.783，P < 0.01；r = 0.793，P < 0.01)。 结论 HPA在糖尿病肾病中的表达升高可能参与了糖尿病肾病蛋白尿的发生。     

金雀异黄素对糖尿病大鼠肾皮质SMemb表达及细胞外基质的作用

目的探讨金雀异黄素(Gen,5,7,4’-三羟异黄酮,又称染料木黄酮)对糖尿病 (DM)大鼠肾组织系膜细胞(MC)表型和细胞外基质的作用。方法雄性SD大鼠45只,分为(1) 正常对照组:10只,普通鼠饲料自由饮食;(2)DM组:再分为4周组9只,8周组8只,普通鼠饲料自由饮食;(3)DM+Gen灌胃组:同样再分为4周组10只,8周组8只,普通鼠饲料自由饮食及 Gen 30 mg．kg-1·d-1。于实验4周、8周末禁食12 h,检测空腹血糖(FBG)、BUN、Scr,肾组织非肌肉型肌球蛋白重链(SMemb)mRNA表达、纤连蛋白(Fn)／肾重及基质金属蛋白酶(MMP)-2／组织基质金属蛋白酶抑制剂(TIMP)-I沉积结果。结果 DM大鼠FBG、BUN、Scr、Fn／肾重均高于正常对照组,以8周时最为显著(P<0．01);MMP-2／TIMP-1免疫组化半定量比值显著降低(P< O．01);SMemb mRNA／GAPDH mRNA半定量比值高于正常对照组,于4周达高峰,8周时下降 (0．794±0．037比0．708±0．029)。Gen干预后与同期DM组相比,肾功能部分恢复,SMemb mRNA／ GAPDH mRNA比值明显缩小(P<0．01);Fn／肾重也显著低于DM组(P<0．01);MMP-2／TIMP-1 比值与同期DM组相比也有显著增加(分别P<0．01或<0．05)。结论金雀异黄素可以减轻链脲佐菌素所诱导DM大鼠的肾皮质SMemb的表达,提高MMP-2／TIMP-1比值,改善糖尿病模型大鼠肾功能,可能有延缓糖尿病肾病(DN)进展的作用。     

法安明对糖尿病大鼠血清及肾组织中P-selectin的作用

目的：探讨P-selectin在糖尿病肾病发病中的作用以及低分子肝素法安明对肾脏保护作用的机制。方法：用链脲佐菌素（STZ60mg/kg体重）构建糖尿病大鼠模型后随机分为糖尿病组（DN组）和法安明治疗组（T组）,并设正常对照组（N组）。于应用法安明前、后第4、8、12周分别测定3组大鼠的体重、24h尿蛋白总量、24h尿白蛋白、Scr,12周测血脂、凝血功能;应用ELISA方法检测各组大鼠血P-selectin水平,用免疫组化方法检测肾组织P-selectin,用real-time PCR法测P-selectin mRNA表达;并观察各组大鼠肾脏组织病理学改变。结果：与N组比较,DN组大鼠肾重/体重指数以及24h尿白蛋白、蛋白定量、Scr均显著上升（P〈0.05）;肾组织P-selectin mRNA和蛋白表达增强（P〈0.01或P〈0.05）。与DN组比较,T组大鼠、肾重/体重指数下降（P〈0.05）、24h尿白蛋白、24h尿蛋白量和Scr均显著下降（P〈0.01）;P-selectin mRNA表达水平显著下降（P〈0.01）。结论：P-selectin可能参与了DN的发病过程,法安明可能通过影响P-selectin的表达从而对肾脏有保护作用。     

IL-18、IL-6、IL-10在糖尿病大鼠肾脏的表达

目的检测IL-18、IL-6、IL-10在糖尿病大鼠肾脏的表达,探讨其在糖尿病肾脏病变过程中的作用。方法将40只雄性Wistar大鼠随机分为正常对照4周组（NC1组）、8周组（NC2组）、糖尿病4周组（DMI组）、8周组（DM2组）,每组10只。DM组给予一次性腹腔内注射60mg／kg链脲佐菌素建立糖尿病大鼠模型。检测各组大鼠体重、肾重、尿微量白蛋白排泄率（UAER）。用免疫组化方法检测肾组织IL-18、IL-6、IL-10的表达,利用计算机图像分析系统进行定量分析。结果DM组大鼠的肾重指数（KWI）、UAER较NC组显著增高;DM2组较DM1组明显增高;IL-18、IL-6在NC组肾组织中仅少量表达,而在DM1组表达明显增多,在DM2组表达增高则更为显著。IL-10在NC组丰富表达,而在DM1组表达明显减弱,在DM2组几乎无表达。肾脏局部IL-18、IL-6与IL-10的表达呈显著负相关。结论糖尿病大鼠肾脏局部IL-18、IL-6表达明显升高、IL-10水平显著降低,IL-18、IL-6与IL-10之间平衡的紊乱在糖尿病肾脏损害病程中发挥重要的作用。