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塑化剂对慢性肾衰大鼠模型和腹膜透析患者腹膜纤维化调控因子TGF-β1/Smads表达的影响 预览

Effect of Plasticizers on the Expression of Peritoneal Fibrosis Regulating Factor TGF-β1/Smads in Chronic Renal Failure Rats and Peritoneal Dialysis Patients
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摘要 目的研究塑化剂对慢性肾衰大鼠模型和腹膜透析患者腹膜纤维化调控因子——转化生长因子-β1(TGF-β1)/Smad蛋白家族(Smads)表达的影响。方法紫外分光光度法测定腹膜透析液中邻苯二甲酸二(2-乙基)己酯(DEHP)的含量。30只雄性SD大鼠随机平均分为正常对照组、模型对照组、低浓度DEHP组、中浓度DEHP组、高浓度DEHP组。模型对照组和DEHP组采用5/6肾切除法制备大鼠肾衰模型,建模成功后每日向大鼠腹腔注射含不同浓度DEHP的"腹透液"连续28d。苏木精-伊红染色法(H-E)、蛋白质印迹法(Western-blot)及反转录PCR(RT-PCR)技术观察大鼠腹膜组织病理、TGF-β1/Smads通路关键因子及基因的表达,免疫荧光法检测大鼠腹膜组织α平滑肌肌动蛋白(α-SMA)的表达。16例腹膜透析患者随机分为2组,分别给予聚氯乙烯(PVC)包装腹透液和非PVC包装腹透液,3月后测量2组腹膜平衡试验、尿素清除指数(KT/V)、肌酐清除率(Ccr)和腹透液中TGF-β1含量。结果 DEHP组表现出腹膜纤维化趋势。与对照组比较,DEHP组TGF-β1/Smads关键因子及其mRNA的表达水平有明显上调(P〈0.05);DEHP组间各蛋白及mRNA的表达比较,差别无统计学意义(P〉0.05)。DEHP组Smad7mRNA和蛋白表达水平较对照组下调(P〈0.05),DEHP各组间比较,差别无统计学意义(P〉0.05)。2组腹膜透析患者KT/V,Ccr及腹膜平衡试验结果比较,差别无统计学意义(P〉0.05);非PVC组腹透液中TGF-β1浓度较PVC组明显降低(P〈0.05)。结论 DEHP可引起慢性肾衰模型大鼠腹膜纤维化的发生,机制为致经典纤维化通路TGF-β1/Smads激活,激活程度与DEHP浓度无关。PVC包装腹膜透析液能上调腹膜TGF-β1的表达,可能导致腹膜纤维化进展。 Objective To investigate the effect of plasticizer on the expression of peritoneal fibrosis regulatory factors,the transforming growth factor-β1/Sma Mad-related protein(TGF-β1/Smad)family in chronic renal failure rat models and peritoneal dialysis(PD)patients. Methods Concentration of diethylhexyl phthalate(DEHP)in peritoneal dialysis fluid was determined by UV spectrophotometric method. 30 male SD rats were randomly divided into the normal control group(Group C),the model control group(Group O),the low-dose DEHP group(Group L),the medium-dose DEHP group(Group M),and the high-dose DEHP group(GroupH). Animals in the model control group and the DEHP groups underwent 5/6 nephrectomy to establish the renal failure model. After the success of modeling,"peritoneal dialysis fluid"with different concentrations of DEHP was intraperitoneally injected into the rats for28 days. The rats were then sacrificed and peritoneal tissue was removed for testing. Hematoxylin-eosin staining(H-E),Western blotting and reverse transcription PCR(RT-PCR)were applied to observe the pathology of rat peritoneum and the protein and mRNA expression of key factors in the TGF-β1/Smads pathway. Expression ofα-Smooth muscle actin(α-SMA)in rat peritoneal tissue was determined by immunofluorescence method. 16 PD patients were randomly divided into 2 groups and assigned to receive dialysis fluid packed either in polyvinyl chloride(PVC)bags or in non-PVC bags. Peritoneal equilibration test,urea clearance index(KT/V),creatinine clearance rate(Ccr),and TGF-β1 content in peritoneal lysate were detected after 3 months. Results A tendency of peritoneal fibrosis was noted in the DEHP groups. Compared with the control groups,protein and mRNA levels of the key factors in the TGF-β1/Smads pathway were significantly increased in the DEHP groups(P〈0.05),and no statistically significant differences in the protein and mRNA levels were observed between the DEHP groups(P〈0.05). Expression l
作者 罗来敏 张桂玲 LUO Laimin;ZHANG Guiling(Department of Nephrology,The First Affiliated Hospital of Nanchang University, Nanchang 330006, China)
出处 《福建医科大学学报》 2018年第2期90-96,134共8页 Journal of Fujian Medical University
基金 江西省教育厅科学技术研究项目(14042)
关键词 邻苯二醛类 腹膜透析 腹膜后纤维化 信号处理 计算机辅助 转化生长因子Β phthalic anhydrides peritoneal dialysis retroperitoneal fibrosis signal processing computer-assisted transforming growth factor beta
作者简介 罗来敏,男,副主任医师,医学硕士.Email:11m090626@163.com
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