转化生长因子β1诱导肾小管上皮细胞转分化的蛋白质组学研究

目的 研究转化生长因子（TGF）β1诱导肾小管上皮细胞（NRK52E）转分化（EMT）过程中相关靶蛋白的表达变化。 方法 应用比较蛋白质组学方法。用双向凝胶电泳（2-DE）对TGF-β1刺激组和对照组 NRK52E细胞总蛋白进行分离。两组间差异蛋白点用质谱和数据库搜索鉴定，并用RT-PCR和Western 印迹在蛋白质和mRNA水平上进一步检测验证。 结果 鉴定出22个差异蛋白，包括与细胞骨架相关蛋白、参与物质转化和能量代谢的酶类、与蛋白翻译后修饰相关蛋白、细胞因子及其他蛋白等。应用RT-PCR验证了转胶蛋白（transgelin）、ATP合成酶α亚型、苹果酸脱氢酶、丝氨酸（半胱氨酸）蛋白酶抑制因子、泛素结合酶9（Ubc9）和表皮生长因子8在mRNA水平的表达差异，与2-DE结果一致。用Western 印迹验证了transgelin、ATP 合成酶α亚型两种蛋白的表达差异，亦与2-DE结果一致。 结论 TGF-β1刺激NRK52E细胞EMT发生过程中可诱导包括与细胞骨架相关蛋白、与翻译后修饰相关蛋白、代谢酶类及细胞因子等多种蛋白表达变化。本研究结果有助于深入理解EMT的具体分子机制及阐明肾脏疾病慢性进展的机制。     

依那普利对梗阻性肾病大鼠肾组织p38丝裂素激活蛋白激酶活性的影响

目的研究血管紧张素转换酶抑制剂(ACEI)依那普利对梗阻性肾病模型肾组织p38丝裂素激活蛋白激酶(MAPK)活性的影响。方法采用单侧输尿管结扎(UUO)模型,治疗组从造模前24h至造模后28d以依那普利10mg·kg-1·d-1灌胃,另设假手术组作正常对照。分别于造模后1h,3h,6h,12h,1d,3d,5d,7d,14d,21d及28d取肾组织,应用免疫沉淀结合特异性底物磷酸化法测定肾组织p38MAPK活性;免疫组织化学法检测磷酸化p38MAPK在肾组织中的表达和定位;免疫组织化学及原位杂交方法检测肾组织TGF-β1mRNA和蛋白水平的表达。结果正常大鼠肾组织基础的p38MAPK活性(吸光度值)为0.22±0.06。UUO术后1h,p38MAPK即被激活(0.45±0.14,P<0.01),并呈进行性升高,12h时达第1个高峰(0.91±0.07,P<0.01),此后活性逐渐下降;第3天后又进行性升高,第7天达到第2个高峰(0.93±0.06,P<0.01)。TGF-β1的表达在UUO术后1h、3h、6h、12h及24h均无明显增加;在第3天有明显增加(A值,13.55±6.33比基础4.32±1.72,P<0.01);第7天达高峰(26.78±8.77,P<0.01)。梗阻肾肾组织p38MAPK的激活明显早于TGF-β1表达,且p38MAPK早期活性的强弱与肾组织TGF-β1的表达水平相一致。依那普利治疗可以明显抑制p38MAPK活性(下降36%～65%,P<0.01),显著降低肾组织TGF-β1表达(下降33%～4     

终末期肾脏病患者血清调脂素(Leptin)水平与营养不良的关系

目的 :探讨终末期肾脏病 (ESRD)患者血清Leptin水平改变与营养不良的关系及意义。 方法 :应用酶联免疫法 (ELISA)及放射免疫法测定 40例ESRD患者和 12例健康志愿者血Leptin及前白蛋白、白蛋白、转铁蛋白等水平 ,同时应用临床上常用的整体评估 (subjectiveglobalassessment ,SGA )作营养不良分级 ,并对血Leptin水平与营养不良等级之间进行Spearman等级相关分析。结果 :ESRD患者Leptin水平明显较正常对照组高 (39.4± 7.3ng/mlvs18.4± 3 .3ng/ml,(P <0 .0 5 ) ,并与SGA营养不良等级呈正相关。 结论 :ESRD患者血清Leptin异常升高与其营养不良状态密切相关 ,可能是引起ESRD患者厌食、蛋白质摄入不足、能量消耗增多的因素之一。     

来氟米特治疗难治性肾病综合征60例临床研究

目的观察长程应用来氟米特联合泼尼松治疗难治性肾病综合征患者的疗效和安全性。方法2000-11～2003-05收集中山大学附属第三医院的难治性肾病综合征病人60例,随机分为两组,分别为接受来氟米特联合泼尼松治疗组(观察组),接受吗替麦考酚酯联合泼尼松治疗组(对照组)。观察治疗2、4、6、8、12、16、20、24、28周的相关临床指标变化及其评价。结果来氟米特组治疗后24h尿蛋白定量显著减少(P<0.05),血清白蛋白显著升高(P<0.05),完全缓解率为56.67%,总有效率为80%。与吗替麦考酚酯组比疗效差异无显著性(P>0.05),来氟米特组12周内显效率高于吗替麦考酚酯组(P<0.05),不良反应轻微,所有病人耐受性良好。结论来氟米特能够有效缓解难治性肾病综合征。     

重视慢性肾脏疾病患者的血压控制

流行病学调查显示 ,慢性肾炎高血压的发生率为 60 %～80 %,其中硬化性肾炎的发生率达 90 %,终末期肾衰竭的发生率在 95 %以上 ,高血压可以在相当程度上影响着慢性肾脏疾病患者心血管事件 (ＣＶ)的发病率和致死率 ,同时也是加速慢性肾衰竭进展的最重要危险因素。对于慢性肾脏疾病患者来说 ,将血压控制至靶目标值可以减少ＣＶ事件的发生及维持肾功能。所以 ,慢性肾脏疾病患者的治疗主要集中在控制血压 ,减少蛋白尿及实现这些目标所需要的治疗手段。在ＭＤＲＤ(ｍｏｄｉｆｉｃａｔｉｏｎｏｆｄｉｅｔｉｎｒｅｎａｌｄｉｓｅａｓｅｓ)的研究中发…     

IgA肾病患者血清algA1对系膜细胞增殖的影响

目的 观察Iga肾病(IgAN)患者与正常人的血清热聚合IgA1(algA1)对系膜细胞(MC)增殖的影响.方法 收集原发性IgAN患者及健康人的血清标本,利用Jacalin-agarose亲和层析联合Sephacryl S-200 HR分子筛层析法分离获得血清单体IgA1(mlgA1),将mlgA1热聚合为algA1.利用患者及正常人的algA1分别刺激小鼠MSC-1097系膜细胞株,利用MTT法检测algA1对系膜细胞增殖的影响.结果 algA1刺激48h组的吸光度是24h组的1.59倍(P=0.000).不同浓度algA1刺激组问MTT吸光度的差异有统计学意义(P=0.000),相比于阴性对照组(SFM).0.25 mg/mL组、0.5 mg/mL组、1 mg/mL组的吸光度分别为SFM的1.67、1.74、1.77倍(P=0.000);2 mg/mL组的吸光度是SFM的1.15倍(P=0.086).在刺激24h时患者0.25 mg.mL组的algA1就可促进系膜细胞增殖了,其吸光度是SFM的1.34倍(P<0.05),而正常对照组则耍在浓度达到1mg/mL时才引起系膜细胞增殖,其吸光度为SFM的1.33倍(P<0.05).结论 研究结果提示患者及正常人的algA1均可促进系膜细胞增殖,这种增殖表现为时间依赖性及一定的剂量依赖性.在高浓度时algA1促系膜细胞增殖的作用消失了;患者的algA1可能更易与系膜细胞结合,刺激相同时间时患者低浓度algA1既可引起MC增殖.     

Rosiglitazone down-regulates lipopolysaccharide-induced expression of CD40 and intercellular adhesion molecule 1 in rat peritoneal mesothelial cells through a NF-κB dependent mechanism

Objective To investigate the effect and mechanism by which PPARγ ligand, rosiglitasone, regulates the expression of CD40 and intercellular adhesion molecule 1 (ICAM-1) in the rat peritoneal mesothelial cells (RPMCs) induced by lipopolysaccharide (LPS). Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were randomly divided into groups as follows: medium, LPS (5 mg/L), LPS (5 mg/L)+BAY11-7085(5 μmol/L, NF-κB inhibitor), rosiglitazone (10 μmol/L or 20 μmol/L, peroxisome proliferator-activated receptor γ activator), LPS (5 mg/L)+rosiglitazone (10 μmol/L)+GW9662 (3 μmol/L, peroxisome proliferator-aetivatcd receptor γ antagonist), and LPS (5 mg/L)+vehicle (DMSO 0.2 ml/L). The expressions of CD40 and ICAM-1 RNA in RPMCs were examined by RT-PCR after 3 hour treatment, and the protein expressions of CD40, ICAM-1, p-NF-κB p65 and p-IκBα were examined by Western blot or immunofluorescence after 24 hour treatment. Results Following treatment with LPS, both the expressions of CD40 and ICAM-1 protein in RPMCs were up-regulated significantly (P<0.05), and the phosphoralation of p65 was increased greatly (1.10±0.17 vs 0.55±0.06, P<0.05). BAY11-7085 (5 μmol/L) significantly decreased the protein expression of p-p65 (0.22±0.11 vs 1.10±0.17, P<0.01), CD40 (0.34±0.02 vs 0.50±0.06, P<0.05) and ICAM-1 (0.35±0.16 vs 0.74±0.03, P<0.05). Pretreated with rosiglitazone for 3 h then added with LPS for 1 h, the levels of p-p65, CD40 and ICAM-1 in RPMCs were significantly decreased compared with those of LPS group (0.77±0.08 vs 0.90±0.10, P相似文献   

应重视和加强大肠杆菌所致腹膜透析相关腹膜炎的防治

持续性不卧床腹膜透析(CAPD)临床应用已30年,患者生存及技术存活显著改善.     

透出液蛋白丢失与连续性非卧床腹膜透析患者营养不良-炎性反应-动脉硬化综合征的相关性

Objective To investigate the impact of peritoneal albumin leakage on malnutrition-inflammation-atherosclerosis (MIA) syndrome in continuous ambulatory peritoneal dialysis (CAPD) patients. Methods A cross-sectional study of a cohort of 130 CAPD patients without edema or active infection was performed. In order to identify peritoneal transport characteristics in CAPD patients, a standard peritoneal equilibration test (PET) was carried out. For malnutrition and inflammation, serum albumin and high-sensitivity C-reactive protein (hs-CRP) levels were measured. Mean-carotid artery intima media thickness (IMT) was used to determine atherosclerosis. Residual glomerular filtration rate (rGFR) was defined as the average of 24-hour urinary urea and creatinine clearances. Results Pearson and Spearman correlation analysis showed that peritoneal albumin leakage amount was positively correlated with age, body mass index, night dwell time, blood glucose, 4 h D/P creatinine levels and hs-CRP levels (r=0.204, P<0.05 ;r=0.314, P<0.01; r=0.265, P<0.01; r=0.212, P<0.05; r=0.401, P<0.01; r=0.216, P<0.05); whereas it was negatively correlated with diastolic perssure, serum albumin levels, glucose level of dialyzate and peritoneal Kt/V (r=-0.209, P<0.05; r=-0.123, P<0.05; r=-0.271, P<0.01; r=-0.212, P<0.01). Overall, there was no correlation between peritoneal albumin leakage and IMT. Patients was significantly greater (P<0.01), and there was a positive correlation between peritoneal albumin leakage amount and IMT (r=0.650, P<0.01). Conclusions Peritoneal albumin leakage is significantly associated with peritoneal transport characteristics, malnutrition and inflammatory state in CAPD patients. High peritoneal albumin leakage amount is a risk factor for atherosclerosis in patients with rGFR less than 1 ml·min-1(1.73 m2)-1.