骨形态蛋白-7在早期糖尿病肾病大鼠中的表达  

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

作　　者：刘道勤[1] 张道友[1] 武其文[2] 

机构地区：[1]皖南医学院第一附属医院肾脏科,安徽241001 [2]皖南医学院第一附属医院检验科,安徽241001

出　　处：《中国血液净化》2016年第2期95-99,共5页Chinese Journal of Blood Purification

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 m RNA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 m RNA(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。Objective To investigate the expression of bone morphogenetic protein- 7 （BMP-7） in rat early 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 statisti- cally 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, P 0.01） 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 no

关 键 词：骨形态蛋白-7 糖尿病肾病 表达 大鼠模型 

分 类 号：R692.5[医药卫生—泌尿科学]