巢蛋白在阿霉素肾病模型大鼠肾组织中表达及霉酚酸酯的干预作用

背景：研究证明,足细胞损伤是产生肾小球性蛋白尿的重要机制。而关于足细胞内众多骨架蛋白如何互相调节维持足细胞特有的形态目前尚未完全了解,足细胞骨架的构建和重塑也成为蛋白尿发生机制的研究热点。 目的：构建阿霉素微小病变肾病大鼠模型,以霉酚酸酯进行干预,检测大鼠肾组织中巢蛋白(nestin)的表达。 方法：纳入雄性Wistar大鼠36只,随机均分为肾病模型组、霉酚酸酯组、正常组(n=12)。肾病模型组、霉酚酸酯组大鼠一次性尾静脉注射阿霉素进行造模,正常组尾静脉注射等量生理盐水。霉酚酸酯组大鼠于造模次日给予霉酚酸酯灌胃,20 mg/(kg•d),1次/d;其他两组大鼠每日给予等量生理盐水。各组分别于造模后14,21,28 d各处死4只大鼠,取肾皮质进行苏木精-伊红染色和免疫组化染色,观察大鼠肾组织病理学改变以及nestin表达情况。 结果与结论：正常组大鼠肾小球滤过膜结构完整,上皮细胞足突清晰;肾病模型组大鼠肾小球上皮细胞足突广泛融合,基底膜正常;霉酚酸酯组大鼠肾小球上皮细胞足突部分融合,但病变较轻。免疫组化结果提示从造模第14天开始,肾病模型组和霉酚酸酯组大鼠nestin表达明显增加,与正常组比较差异有显著性意义(P < 0.05);霉酚酸酯组大鼠nestin表达低于肾病模型组,差异有显著性意义(P < 0.05)。提示肾小球足细胞损伤时,足细胞内nestin表达增多,随病情加重而表达增高。霉酚酸酯可以减轻足细胞损伤,下调nestin的表达,维持足细胞的正常结构,达到延缓肾脏损害的目的。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程      

多囊卵巢综合征大鼠模型诱导的实验研究

目的建立多囊卵巢综合征（PCOS）大鼠模型。方法通过皮下埋置17-炔诺酮硅胶棒联合皮下注射HCG（Bogovich法）诱导PCOS大鼠模型。测定血清睾酮（T）空腹血糖（FPG）及胰岛素（FIns）水平。结果模型组胰岛素抵抗指数（Homa-IR）、FPG、Fins及T水平均高于对照组,24dPCOS大鼠模型构建成功。结论 Bogovich法诱导的PCOS大鼠模型存在高雄激素水平及代谢异常,是研究PCOS发病机制的理想动物模型。     

蝮蛇抗栓酶治疗肾病综合征疗效观察

本文报告蝮蛇抗栓酶分别与激素,免疫抑制剂不同组合治疗原发性肾病综合征72例,比较单用激素,免疫抑制剂及加用蝮蛇抗栓酶组合后的疗效。与激素组比较X~2=5.3814,P<0.05,与激素、CTX组比较X~2=4.555,P<0.05。加酶组的疗效具有明显的优越性。故认为蝮蛇抗栓酶是一种治疗肾病综合征有前途而又安全经济的药物。     

蝮蛇抗栓酶、强的松治疗肾病综合征高凝状态的对照

蝮蛇抗栓酶、强的松治疗肾病综合征高凝状态的对照文／周辽军肾病综合征患者往往出现高凝状态，其血液粘滞性与正常人不同。本文合并应用蝮蛇抗栓酶（Ａｈａｌｙｓａｎｔｉｎｆａｒｃｔａｓｅ；批号：９２０３０２１）与强的松（Ｐｒｅｄｎｉｓｏｎｕｍ；批号：９２１００...     

小儿肾病综合征血管紧张素I转换酶基因多态性研究

目的探讨ACE基因第16内含子插入/缺失多态性与肾病综合征、血清ACE活性的关系. 方法采用聚合酶链式反应(PCR)检测82例肾病综合征患儿和38例正常儿童ACE基因并同时用酶偶联法测血清ACE活性. 结果①82例肾病综合征患儿中ACE基因II型,ID型和DD型频率分别为47.6%(39/82),24.4%(20/82)和28%(23/82).对照组分别为47.3%(18/38),23.7%(9/38)和29%(11/38),肾病综合征患儿和正常儿童之间ACE基因I/D多态性频率差异无显著性意义(p>0.05).②38例正常儿童和82例肾病综合征患儿血清ACE活性比较差异无显著性意义(p>0.05),ACE各基因型中血清ACE活性差异有显著性意义(p<0.01),DD型>ID型>II型. 结论①小儿肾病综合征ACE基因II型,ID型和DD型频率分布和正常儿童差异无显著性意义(p>0.05).②ACE基因多态性与血清ACE活性密切相关.     

依托咪酯对新生大鼠皮质神经元钙激活性钾通道的作用

采用膜片钳技术的吸附式和内面向外式两种构型研究了依托咪酯对增减的ＳＤ新生鼠皮持 钙激活性钾（ＫＣａ）通道的作用。结果：（１）新生ＳＤ大鼠皮质神经元上存在有单元电导为１７６．６５±３７．９７ｐＳ的ＫＣａ通道;（２）依托咪酯０．８、４、１０、２０四个不同剂量在吸附式构型时浓度依赖性激活ＫＣａ通道,其开放概率较对照组均明显增大,（３）依托咪酯０．８μｍｏｌ／Ｌ在内面向外式构型时对ＫＣａ通道也有明显激活作     

小儿肾病综合征血管紧张素I转换酶基因多态性研究

目的：探讨ACE基因第16内含子插入／缺失多态性与肾病综合征、血清ACE活性的关系。方法：采用聚合酶链式反应（PCR）检测82例肾病综合征患儿和38例正常儿童ACE基因并同时用酶偶联法测血清ACE活性。结果：①82例肾病综合征患儿中ACE基因Ⅱ型，ID型和DD型频率分别为47．6％（39／82），24．4％（20／82）和28％（23／82）。对照组分别为47．3％（18／38），23．7％（9／38）和29％（11／38），肾病综合征患儿和正常儿童之间ACE基因I／D多态性频率差异无显著性意义（P＞0．05）。②38例正常儿童和82例肾病综合征患儿血清ACE活性比较差异无显著性意义（P＞0．05），ACE各基因型中血清ACE活性差异有显著性意义（P＜0．01），DD型＞ID型＞Ⅱ型。结论：①小儿肾病综合征ACE基因Ⅱ型，ID型和DD型频率分布和正常儿童差异无显著性意义（P＞0．05）。②ACE基因多态性与血清ACE活性密切相关。     

麦考酚钠肠溶片与吗替麦考酚酯胶囊联合质子泵抑制剂对肾移植患者酶酚酸暴露的影响

背景：麦考酚钠肠溶片与经典的免疫抑制剂吗替麦考酚酯胶囊在体内吸收代谢场所有所不同,与相关药物的联合应用造成了二者在代谢水平的差异,从而可能导致临床疗效的差异。 目的：探讨麦考酚钠肠溶片与吗替麦考酚酯胶囊联合质子泵抑制剂的临床应用对肾移植患者酶酚酸暴露的影响。 方法：选取2010-05/2011-05于解放军第309医院全军器官移植中心应用麦考酚钠肠溶片患者17例和应用吗替麦考酚酯胶囊患者16例为研究对象,采用麦考酚钠肠溶片/吗替麦考酚酯胶囊+环孢素/他克莫司+类固醇激素的免疫抑制方案,麦考酚钠肠溶片540 mg 12 h给药1次,吗替麦考酚酯胶囊750 mg 12 h给药1次。 结果与结论：麦考酚钠肠溶片组和吗替麦考酚酯胶囊组患者肾移植后肾功能均恢复正常。麦考酚钠肠溶片组酶酚酸平均达峰时间、平均达峰浓度、平均半衰期和均平均药-时曲线下面积高于吗替麦考酚酯胶囊组(P < 0.05),说明麦考酚钠肠溶片临床联合应用质子泵抑制剂对肾移植患者酶酚酸暴露的治疗效果优于吗替麦考酚酯胶囊。     

活性氧在阿霉素肾病中致病作用的实验研究

本实验用阿霉素一次静注的方法,制作了大鼠肾病综合征的模型。大鼠肾皮质过氧化脂质丙二醛(MDA)在第8天达到高峰,为27.16±4.36nmol/100mg,血清MDA也明显升高。用活性氧清除剂超氧化物歧化酶(清除O_2~(·-))、过氧化氢酶(清除H_2O_2)、二甲基亚砜(清除·OH)治疗阿霉素肾病,可见肾皮质MDA显著减少(P<0.01),血清MDA也显著减少(P<0.05～0.01),蛋白尿明显降低(P<0.01),肾脏病理变化也减轻。由此可见,活性氧在阿霉素肾病的发病机制中起重要作用。     

糖尿病肾病大鼠动物模型建立及INF-α干预机制研究

目的 探讨糖尿病肾病大鼠动物模型建立及TNF-α干预机制。方法 选择 SD大鼠60只作为对象,随机数字法分为对照组和观察组,各30只。对照组大鼠腹腔注射柠檬酸缓冲液,观察组在对照组基础上采用高糖高脂饲料联合5%葡萄糖饮水喂养,诱导胰岛素抵抗,2周后腹腔注射STZ,连续注射3 d,诱发持续高糖血症。采用酶联免疫吸附法在建模前、建模2周、建模4周检测观察组大鼠血糖、尿白蛋白、肌酐水平,同时检测两组大鼠TNF-α水平,分析糖尿病动物模型建立及TNF-α干预机制。结果 ①对照组大鼠全身状态良好,体质量增加,皮毛光滑亮泽,观察组大鼠建模成功后出现多食、多饮、多尿及生长迟缓现象,成模4周后体重低于对照组,差异有统计学意义（P＜0.05）;②观察组建模2周血糖水平（8.21±0.21）mmol/L,建模4周血糖水平（31.21±1.04）mmol/L,均高于对照组的（6.32±0.19）mmol/L、（6.74±0.24）mmol/L）,差异有统计学意义（P＜0.05）;③观察组建模2周TNF-α水平为（3.03±0.23）mmol/L,建模4周TNF-α水平为（12.10±1.12）mmol/L,均高于对照组的（1.64±0.58）mmol/L、（1.69±0.62）mmol/L,差异有统计学意义（P＜0.05）。结论 高糖高脂饲料联合浓度为5%葡萄糖饮水配合小剂量STZ能建立理想的糖尿病肾病大鼠动物模型,可以明确糖尿病肾病发展伴有TNF-α炎症因子水平升高,进一步为TNF-α干预机制研究奠定了基础。     

霉酚酸酯对蛋白超载肾病大鼠p38丝裂素活化蛋白激酶信号转导通路的影响

目的 探讨霉酚酸酯(MMF)对蛋白超载肾病大鼠p38丝裂素活化蛋白激酶(p38MAPK)信号转导通路的影响.方法 SD大鼠30只分3组(n=10):对照组(生理盐水)、BSA组[牛血清白蛋白(BSA)超载肾病大鼠]、治疗组(BSA+MMF).采用蛋白超载肾病大鼠模型,于4周后观察各组大鼠尿素氮(BUN)、血肌酐(Scr)、24h尿蛋白定量;免疫组织化学检测核因子κB(NF-κB),应用凝胶迁移率变动分析检测NF-κB的活性变化;Western免疫印迹分析法检测p38 MAPK的磷酸化水平.光镜和电镜观察大鼠肾组织形态改变.结果 光镜和电镜显示BSA组大鼠出现肾间质淋巴、单核细胞浸润、肾小管萎缩和纤维化等病变;治疗组病变较BSA组明显减轻.与对照组比较,BSA组大鼠24 h尿蛋白增加[(48.3±3.3)mg/24h比(67.8±4.5)mg/24 h,P<0.05],NF-κB和p38 MAPK表达增加(45.24±6.25比88.59±7.43和80.68±8.34比235.23±10.41,P<0.05),且p38 MAPK表达与NF-κB和24h尿蛋白定量呈正相关(r=0.72,r=0.65,P<0.05).与BSA组比较,治疗组MMF可减少尿蛋白排泄[(59.1±4.2)mg/24 h,P<0.05],同时抑制p38 MAPK磷酸化和NF-κB的表达(P<0.05).结论 MMF可减少NF-κB的表达,其机制可能依赖于抑制p38 MAPK的磷酸化.     

Influence of mycophenolate mofetil （MMF） upon the maturation and allo-stimulatory activity of cultured progenitors of dendritic cells and the effects on the tolerance induction in allograft recipients

Dendritic cells (DCs) are highly potent antigenpresenting cells of bone marrow origin that canstimulate both primary and secondary T and B cell responses [1]. These cells have every neces sary component for potent antigen presenting functions, including the production of a variety of im portant immunostimulatory cytokines and the ex pression of critical cell surface molecules. De pending on their level of maturity, DCs expressprominent levels of MHC classⅠand classⅡm…     

霉酚酸酯对糖尿病大鼠肾脏的保护作用

目的探讨霉酚酸酯(MMF)对糖尿病模型大鼠肾脏的保护作用及其机制。方法SD大鼠随机分为3组:对照组、糖尿病模型组和糖尿病治疗组。治疗12周后,检测大鼠血糖(BG)、血尿素氮(BUN)、血肌酐(Scr)、肾肥大指数(肾重KW/体重BW)、肌酐清除率(Ccr)和24h尿蛋白(24Upro)。肾组织做常规光镜和电镜检查。用免疫组织化学方法检测肾组织中细胞间黏附分子-1(ICAM-1)和增殖细胞核抗原(PCNA)蛋白的表达,半定量RT-PCR的方法检测肾组织中ICAM-1mRNA的表达。结果与对照组相比,糖尿病组大鼠BG、KW/BW、24Upro、BUN、Scr和Ccr均显著升高(P<0.05或0.01);系膜密度和肾小球基底膜厚度均显著增加(P<0.01);肾组织内ICAM-1和PCNA的蛋白质表达及ICAM-1的mRNA表达均显著上调(P<0.05或0.01)。除血糖外,治疗组上述指标均显著回降(P<0.05或0.01)。结论MMF对糖尿病肾病有保护作用,其机制可能与下调ICAM-1和PCNA的表达有关。     

吗替麦考酚酯联合来氟米特多靶点诱导治疗狼疮肾炎的临床疗效

目的评估吗替麦考酚酯（MMF）联合来氟米特（LEF）多靶点诱导治疗狼疮肾炎疗效及安全性。方法选取2010年5月至2013年6月惠州市中心人民医院经肾活检确诊为Ⅲ型、Ⅳ型、Ⅲ＋Ⅴ型、Ⅳ＋Ⅴ型狼疮肾炎且慢性指数（CI）〈4分、年龄大于18岁患者70例,随机分为多靶点治疗组（MMF1．0g／d,LEF10mg／d,共6个月）及环磷酰胺冲击治疗对照组[IV．CYC（0．5～1．0）g／m^2,每月1次,共6个月],每组35例。两组均接受起始剂量甲基泼尼松龙500mg冲击治疗3d,后泼尼松0．5mg·kg^-1·d^-1口服维持4周并逐渐减量,评价肾脏完全缓解（CR）及部分缓解（PR）以及系统性红斑狼疮疾病活动评分（SLEDAI）、抗双链DNA抗体（ds-DNA）值的变化。分析比较两组胃肠道反应、上呼吸道感染、中性粒细胞减少、月经紊乱、带状疱疹等主要不良反应的发生情况。结果共有62例患者纳入统计,其中多靶点治疗组33例,对照组29例,多靶点治疗组CR及CR或PR率与对照组比较,差异无统计学意义。多靶点治疗组Ⅳ＋Ⅴ型CR率高于对照组（44．4％比12．5％,P=0．026）,多靶点治疗组Ⅲ＋Ⅴ型CR率高于对照组（60％比20锈,P〈0．05）,两组Ⅳ＋Ⅴ型PR或CR率比较差异亦有统计学意义（77．8％比50％,P=0．018）。多靶点治疗组SLEDAI评分、ds-DNA变化与对照组比较差异无统计学意义。多靶点治疗组与对照组比较,胃肠道反应及上呼吸道感染发生率低于对照组（6．1％比48．3％,P=0．032;9．1％比24．1％,P=0.041）。两组均出现2例中性粒细胞减少,对照组还出现月经紊乱2例（6．9％）,带状疱疹2例（6．9％）。结论多靶点诱导狼疮肾炎疗效及安全性均优于传统治疗方案。     

霉酚酸酯对糖尿病大鼠肾小管-间质损伤的保护作用及机制

研究霉酚酸酯(MMF)对糖尿病大鼠肾小管-间质损伤的保护作用,并探讨其可能机制。应用链脲佐菌素诱导大鼠糖尿病模型,随机分三组:对照组、模型组与MMF给药组。8周后观察尿白蛋白排泄率(AER)、肾小管-间质病理形态学变化,应用免疫组化方法检测肾小管-间质骨桥蛋白(OPN)与α-平滑肌肌动蛋白(-αSMA)蛋白表达,Western印迹检测肾组织IL-1与TNF-α蛋白表达。结果表明,糖尿病大鼠AER与肾小管-间质损伤指数(TII)明显高于对照组(P<0.01),MMF给药组AER与TII明显低于模型组(P<0.05);免疫组化显示模型组肾小管-间质OPN与-αSMA表达阳性面积明显高于对照组(P<0.01),MMF给药组肾小管OPN与与-αSMA表达阳性面积明显低于模型组(P<0.01);Western印迹显示模型组肾组织IL-1与TNF-α表达较对照组分别增加2.8倍与3.8倍,MMF给药组IL-1与TNF-α表达分别下降73%与70%(P<0.01)。研究显示MMF对糖尿病肾小管-间质损伤有明显保护作用,其机制可能部分与抑制肾小管-间质OPN与-αSMA表达有关。     

Renoprotective effects of combination of angiotensin converting enzyme inhibitor with mycophenolate mofetil in diabetic rats

Objective and design Previously it was shown that blocking of the renin-angiotensin system (RAS) by angiotensin converting enzyme (ACE) inhibitors, or suppression of inflammatory responses by immunosuppressive drugs such as mycophenolate mofetil (MMF) could attenuate renal injury in diabetic rats. Whether RAS blockade combined with an immunosuppressive drug provides superior renoprotection against diabetic nephropathy has not been clearly delineated. Materials Diabetes was induced by injection of streptozotocin after uninephrectomy. Treatment Rats were randomly separated into five groups: control, diabetes, diabetes treated with enalapril (an ACE inhibitor, 10 mg/kg/d by gastric gavage), diabetes treated with MMF (10 mg/kg/d by gastric gavage), or diabetes treated with a combination of both agents and were followed for 8 weeks. Methods 24 h urinary albumin excretion rate (AER) was determined, renal injury was evaluated, immunohistochemistry for ED-1 macrophage marker, intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) were performed, and expression of transforming growth factor (TGF)-β1 protein was determined by Western blotting analysis. Results Diabetes was associated with a considerable increase in AER. Both enalapril and MMF retarded the increase in albuminuria, which was nearly completely abrogated by combination therapy. Increased glomerular volume and tubulointerstitial injury index in diabetic rats was attenuated by treatment with either enalapril or MMF and further reduced by the combination of the two. Elevated malondialdehyde levels in renal tissue were reduced by enalapril or MMF and, more effectively, by combined enalapril with MMF. Renal overexpression of ICAM-1 was not retarded by enalapril and attenuated by MMF or combined enalapril with MMF. Combination therapy was associated with a superior suppression of diabetes-induced macrophage recruitment and overexpression of MCP-1 and TGFβ1 compared to either monotherapy in renal tissue. Conclusion The combination of enalapril and MMF confers superiority over monotherapy in renoprotection, a mechanism which may be at least partly correlated with synergistic suppression of increased macrophage recruitment and overexpression of MCP-1 and TGF-β1 in renal tissue in diabetic rats. Received 26 July 2005; returned for revision 5 October 2005; returned for final revision 9 January 2006; accepted by M. Katori 23 January 2006     

Renal glycosuria in patients with the nephrotic syndrome

Summary Transient or persistent renal glycosuria may occur in patients with the nephrotic syndrome. In an attempt to elucidate its mechanism, glucose titration experiments were performed in 20 nephrotic patients. The type A titration curve was found in one and type B in four patients with severe organic changes and low glomerular filtration rate. The remaining subjects displayed a particular type of curve (type C) characterized by a low point of splay but an otherwise almost physiological tracing. In type B and C patients the maximal rate of reabsorption per ml glomerular filtrate (Tm_G/GFR) was significantly increased and correlated inversely with the filtration fraction. In these patients the point of splay correlated with the glomerular filtration rate and the sodium clearance, but not with the plasma albumin concentration or the rate of proteinuria. These observations suggest that type A was due to diffuse tubular atrophy, and type B to increased nephron heterogeneity resulting from chronic organic changes. Type C was presumably caused by a potentially reversible alteration of the late proximal or distal glucose transport related to the nephrotic syndrome itself.     

Improved gastrointestinal symptoms and quality of life after conversion from mycophenolate mofetil to enteric-coated mycophenolate sodium in renal transplant patients receiving tacrolimus

It is reported that a conversion from mycophenolate mofetil (MMF) to enteric-coated mycophenolate sodium (EC-MPS) relieves gastrointestinal (GI) symptom burden and improves health-related quality of life (HRQoL). However, it is unclear whether renal transplant recipients using tacrolimus receive the same benefit from the conversion. In this prospective, multi-center, open-label trial, patients were categorized into two groups by their GI symptom screening. Equimolar EC-MPS (n=175) was prescribed for patients with GI burdens; those with no complaints remained on MMF (n=83). Gastrointestinal Symptom Rating Scale (GSRS) and Gastrointestinal Quality of Life Index (GIQLI) were evaluated at baseline and after one month. Patients and physicians completed Overall Treatment Effect (OTE) at one month. EC-MPS-converted patients had worse GSRS and GIQLI scores at baseline than MMF-continued patients (all P<0.001). Significant improvements in GSRS and GIQLI scores were observed for EC-MPS-converted patients at one month, but MMF-continued patients showed worsened GSRS scores (all P<0.05). OTE scale indicated that EC-MPS patients improved in overall GI symptoms and HRQoL more than MMF patients did (P<0.001). In tacrolimus-treated renal transplant recipients with GI burdens, a conversion from MMF to EC-MPS improves GI-related symptoms and HRQoL.     

GRβ在肾病综合征患儿中的表达及影响因素研究

目的:探讨原发性肾病综合征(INS)患儿外周血单个核细胞(PBMC)内糖皮质激素受体β(GRβ)表达的意义, 以及激素抵抗型肾病综合征患者的血清对GRβ表达的影响。方法:采用免疫细胞化学和蛋白质印迹方法检测INS患儿GRβ免疫阳性PBMC(GRβ+PBMC)数目和PBMC细胞核内GRβ蛋白量表达, 通过体外细胞培养检测激素抵抗型INS患儿的血清对正常对照组儿童和激素敏感型INS患儿GRβ+PBMC数目和PBMC细胞核内GRβ蛋白质表达的影响。结果:激素抵抗型INS患儿GRβ+PBMC细胞数和细胞核内GRβ蛋白质表达均明显高于激素敏感型INS患儿(P<0.01);经激素抵抗型INS患儿的血清体外共培养24h后, 正常对照组和激素敏感型INS患儿GRβ+PBMC细胞数和细胞核内GRβ蛋白表达量均显著增加(P<0.01).结论:INS患儿糖皮质激素抵抗可能与GRβ的表达亢进有关, 而血清中的炎症因子可能是诱导GRβ高表达、进而导致继发性糖皮质激素抵抗的一个重要因素。     

Radioimmunoassay of plasma digoxin in nephrotic syndrome

Summary Plasma digoxin was determined by three different radioimmunoassays in blood samples from 12 patients with hypoalbuminemia before and after increasing the concentration of albumin and TBG. With the Clinical-Assays-[¹²⁵I]-Kit and the Schwarz/Mann-[³H]-Kit reliable digoxin values were obtained even in patients with severe nephrotic syndrome.Mit Unterstützung der deutschen Forschungsgemeinschaft im Rahmen des SFB 89 Kardiologie Göttingen.