中国血液净化

中国血液净化 ›› 2016, Vol. 15 ›› Issue (02): 95-99.doi: 10.3969/j.issn.1671-4091.2016.02.009

• 基础研究 • 上一篇    下一篇

骨形态蛋白-7在早期糖尿病肾病大鼠中的表达

刘道勤1,张道友1,武其文2   

  1. 皖南医学院第一附属医院1肾脏科，2检验科
  • 收稿日期:2015-08-31 修回日期:2015-10-21 出版日期:2016-02-12 发布日期:2016-02-19
  • 通讯作者: 武其文 yjslab@163.com E-mail:liudaoqincom@163.com

Expression of bone morphogenetic protein- 7 in rats with early diabetic nephropathy

  • Received:2015-08-31 Revised:2015-10-21 Online:2016-02-12 Published:2016-02-19

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摘要： 目的探讨骨形态蛋白-7(bone morphogenetic protein-7，BMP-7)基因在糖尿病肾病大鼠中的表达，进一步阐明BMP-7 基因与糖尿病肾病的关系。方法雄性SD 大鼠40 只，随机分为正常对照组，糖尿病肾病4 周组、糖尿病肾病8 周组，糖尿病肾病12 周组。应用链脲佐菌素建立1 型糖尿病肾病动物模型。正常对照组注射相当剂量的柠檬酸缓冲液作为对照。各组大鼠分别于实验4 周、8 周和16 周末称体质量，金属代谢笼收集尿液并记录24h 尿量，测定24h 尿蛋白量。随后处死大鼠，心脏穿刺取血，分离血清。应用日立7600 型全自动生化分析仪检测血肌酐、尿素氮、血糖水平；应用酶联免疫吸附法测定血、尿BMP-7 的水平；采用实时荧光定量PCR 和免疫组化检测BMP-7 mRNA 和蛋白在肾脏中的表达。结果糖尿病肾病4 周组、8 周组，12 周组和正常对照4 组间在体质量(F=18.040，P＜0.001)、肾重/体质量指数(F=70.20，P＜0.01)和24h 尿量(F=249.6，P＜0.001)的差异均具有统计学意义。4 组间在血糖(F=41.240，P＜0.001)、24h 尿蛋白(F=18.980，P＜0.01)、血肌酐(F=12.690，P＜0.01) 和血尿素氮(F=14.570，P＜0.001)的差异具有统计学意义。4 组间在血BMP- 7(F=1234.380，P＜0.001)、尿BMP- 7(F=
34.510，P＜0.001)、肾脏BMP-7 表达面积(F=70.24，P＜0.01)上的差异均具有统计学意义。与正常对照组相比，糖尿病肾病12 周组的血BMP-7(q 值=75.185，P＜0.01)、尿BMP-7(q 值=13.389，P＜0.01)浓度显著下降，在肾脏中其BMP-7 mRNA(q=8.402，P＜0.01)和蛋白质的表达量(q =17.902，P＜0.01)也显著下降。结论BMP-7 基因在实验性1 型糖尿病肾病大鼠模型早期表达下调。BMP-7 可能参与了糖尿病肾病的发生发展，并可能成为早期诊断糖尿病肾病肾功能损伤的分子标记。

关键词: 骨形态蛋白-7, 糖尿病肾病, 表达, 大鼠模型

Abstract: Objective To investigate the expression of bone morphogenetic protein -7 (BMP-7) in ratearly diabetic nephropathy model for the elucidation of the relationship between BMP-7 and diabetic nephropathy. Methods Forty male SD rats were randomly divided into normal control group, diabetic nephropathy for 4 weeks group, diabetic nephropathy for 8 weeks group, and diabetic nephropathy for 12 weeks group. Streptozotocin 65mg/kg was given intraperitoneally to establish the rat model of type 1 diabetic nephropathy, and citrate buffer was injected for rats in normal control group. Rats were weighed at 4 weeks, 8 weeks and 16
weeks after the injection. Metal metabolic cage was used to collect urine for 24-hour urine protein (24h-UP) and 24 hours urine output. Rats were then sacrificed. Serum creatinine (Scr), blood urea nitrogen (BUN), and glucose (BG) were assayed in a Hitachi 7600 automatic biochemical analyzer, BMP-7 in blood and urine were measured using enzyme-linked immunosorbent assay, and BMP-7 mRNA in kidney was quantified by realtime PCR, and BMP-7 protein in kidney was detected by immunohistochemistry. Results There were statistically differences in body weight (F=18.040, P＜0.001), kidney weight/body mass index (F=70.20, P＜0.01), 24 h urine volume (F=249.6, P＜0.001), blood glucose (F=41.240, P＜0.001), 24h-UP (F=18.980, P＜0.01), Scr (F=12.690, P＜0.01), and BUN (F=14.570, P＜0.001) among the groups of normal control group, diabetic nephropathy for 4 weeks, diabetic nephropathy for 8 weeks, and diabetic nephropathy for 12 weeks. BMP-7 (F=1234.380, P＜0.001), urinary BMP-7 (F=34.510, P＜0.001), renal BMP-7 expression area (F=70.24, P0.01) were also different among the 4 groups. In diabetic nephropathy for 12 weeks group, blood BMP-7 (q=75.185, P＜0.01), urinary BMP- 7 (q=13.389, P＜0.01), BMP- 7 mRNA in kidney (q=8.402, P＜0.01) and BMP- 7 protein in kidney (q=17.902, P＜0.01) were decreased as compared with those in normal control group. Conclusion The expression of BMP-7 gene decreased in type 1 diabetes nephropathy rat model. BMP-7 may be involved in the development of diabetic nephropathy. BMP-7 may become an early diagnostic marker for renal function impairment in diabetes nephropathy.

Key words: Bone Morphogenetic Protein-7, Diabetic nephropathy, Expression, rat model