水通道蛋白在大鼠慢性环孢素A肾毒性中的表达

发布时间：2018-06-17 04:09

本文选题：慢性CsA肾毒性 + 水通道蛋白　；参考：《延边大学》2014年硕士论文

【摘要】：目的：探讨水通道蛋白(Aquaporin,AQP)在大鼠慢性环孢素A肾毒性肾中的表达。方法：雄性Sprague-Dawley大鼠喂饲低盐饲料(0.05%钠盐)1周后,随机分为两组：1)对照组(n=8)：皮下注射橄榄油(1mL.kg-1.d-1)；2)CsA肾毒性组(n=8)：皮下注射CsA(15mg.kg-1.d-1);监测两组大鼠的体重变化、水摄入量、尿量、肾功能(血尿素氮、血肌酐、内生肌酐清除率)；三色染色(Masson染色)观察肾小管间质纤维化；TUNEL检测法和电子显微镜观察细胞凋亡情况；利用免疫组织化学染色和免疫印迹法分别观察肾内AQP-1、AQP-2、AQP-3在肾脏皮质、髓质中的表达。结果：与对照组相比,CsA肾毒性组大鼠表现为每日水摄入量(24.0±6.0mL vs.15.0±4mL,P0.01).尿量(26±7mL vs.14±6mL,P0.01).血尿素氮(36±7mg/dL vs.15±4mg/dL,P0.01).血肌酐(1.1±0.2mg/dL vs.0.8±0.1mg/dL,P0.01)明显增加,而体重(328±8g vs.340±5g,P0.01)和内生肌酐清除率(0.24±0.04mL/min/100g vs.O.35±0.03mL/min/100g,P0.01)减少；免疫组化法示正常肾组织的近曲小管、髓袢升支细段、直管降支均有AQP-1的表达,尤其沿皮质的刷状缘高表达,而AQP-1在CsA肾毒性组大鼠内髓质、外髓质中的表达明显减少[皮质：(48±6%vs.100±5%P0.01)；髓质：(44±2%vs.54±3%)AQP-2在正常肾组织的集合管细胞高表达,而AQP-2在CsA毒性组大鼠肾组织皮质的表达无变化,在髓质的表达显著减少(40±2%vs.102±4%,P0.01)；AQP-3丰富表达于肾脏集合管细胞基底区,在CsA毒性组AQP-3皮质、髓质表达上均明显减少[皮质：(40±2%vs.100±5%)；髓质：(38±2%vs.100±5%)]。这些水通道蛋白表达的变化与肾小管上皮细胞凋亡数呈负向相关(AQP-1:r=-0.789,P=0.001；AQP-2:r=-0.698,P=0.003; AQP-3:r=-0.515.P=0.041). 结论：在慢性环孢A肾毒性中,水通道蛋白1、2、3在皮质、髓质中的表达明显减少,这种现象可能与CsA肾毒性大鼠尿浓缩功能障碍有关。细胞凋亡是水通道蛋白表达减少的分子机制。
[Abstract]:Aim: to investigate the expression of aquaporin in chronic cyclosporine A nephrotoxic kidney in rats. Methods: male Sprague-Dawley rats were fed with 0.05% sodium salt diet for 1 week, and were randomly divided into two groups: control group (n = 1): subcutaneous injection of olive oil 1 mL 路kg -1 路d -1 and CsA 15 mg 路kg -1 路d -1 路d -1. The body weight, water intake, urine volume of the two groups were monitored. Renal function (blood urea nitrogen, serum creatinine, endogenetic creatinine clearance rate) and renal tubulointerstitial fibrosis (tubulointerstitial fibrosis) were observed by Tunel and electron microscopy. Immunohistochemical staining and Western blotting were used to observe the expression of AQP-1 and AQP-2AQP-3 in renal cortex and medulla respectively. Results: compared with the control group, the rats in CSA nephrotoxicity group showed a daily water intake of 24.0 卤6.0 mL vs.15.0 卤4mL P0.01a. Urine volume was 26 卤7 mL vs.14 卤6 mL P0.01g. Blood urea nitrogen 36 卤7 mg / dL vs.15 卤4 mg / dL P0.01g. Serum creatinine was significantly increased by 1.1 卤0.2 mg / dL vs.0.8 卤0.1 mg / dL P0.01), while the body weight was 328 卤8 g vs.340 卤5g P0.01) and the creatinine clearance rate was 0.24 卤0.04mL / min / 100g vs.O.35 卤0.03mL / P / 100g P0.01.The immunohistochemical method showed that AQP-1 was expressed in the proximal convoluted tubules, the ascending branches of the loop pulposus, and the descending branches of the straight canal, especially along the brush-shaped edge of the cortical cortex. The expression of AQP-1 in the medulla and outer medulla of CSA nephrotoxic rats was significantly decreased [cortical fraction 48 卤6%vs.100 卤5P0.01U; medullary: 44 卤2%vs.54 卤3p; AQP-2 was overexpressed in the collecting duct cells of normal renal tissue, but AQP-2 expression did not change in CSA toxic group. The expression of AQP-3 was significantly decreased in the medulla (40 卤2%vs.102 卤4) and in the basal area of the renal collecting duct cells. In the CSA toxic group, the expression of AQP-3 was significantly decreased [cortex: 40 卤2%vs.100 卤5; medulla: 38 卤2%vs.100 卤5]. These changes in aquaporin expression were negatively correlated with the number of apoptosis in renal tubular epithelial cells. Conclusion: in chronic cyclosporine A nephrotoxicity, the expression of aquaporin 1, 2 and 3 in cortex and medulla is significantly decreased, which may be related to urinary concentration dysfunction in CSA nephrotoxic rats. Apoptosis is the molecular mechanism of decreased expression of aquaporin.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692

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