实时荧光定量PCR法检测糖尿病肾病大鼠肾脏中BMP-2与MGP  

Detection of Bone Morphogenetic Protein-2 and Matrix Gla Protein Gene Expression in Kidney of Diabetic Nephropathy Rats with Real-Time PCR

作　　者：赵安菊[1] 黄颂敏[2] 欧三桃[2] 王里[1] 陈超[1] 

机构地区：[1]贵州省人民医院干医科,贵州贵阳550002 [2]四川大学华西医院肾内科,四川成都610041

出　　处：《贵阳医学院学报》2011年第5期460-464,共5页Journal of Guiyang Medical College

基　　金：贵州省科技厅基金资助项目(黔科合J字[2008]2299号);贵州省卫生厅优秀医学青年科技人才基金项目(gzwkj2010-02-006)

摘　　要：目的:检测糖尿病肾病(DN)大鼠肾脏中骨形成蛋白2(BMP-2)和基质Gla蛋白(MGP)的基因表达量的变化规律,探讨其对糖尿病肾病的影响。方法:设对照组(N组)及糖尿病肾病组(DN组,STZ诱导的DN大鼠模型);采用实时荧光定量PCR(Real-Time PCR)方法检测4、12、24周DN大鼠及正常大鼠肾脏中BMP-2及MGP的基因定量,茜素红染色观察肾实质内小动脉周围钙盐沉积。结果:(1)DN组各时间点血糖及24 h尿蛋白量均较N组显著增高(P<0.05),自第12周开始,DN组血肌酐、血磷较N组明显增高(P<0.05);(2)DN组4周时BMP-2 mRNA已有表达,随时间延长表达逐渐升高,24周最高(P<0.05);DN组MGP表达量4周最高,随时间延长逐渐降低,24周最低(P<0.05),而N组BMP-2和MGP mRNA各时间点表达量差异均无统计学意义;(3)茜素红染色DN组第4～12周时偶见肾内小动脉少许点状红色钙盐沉积,第24周时可见肾实质内小动脉及周围组织有较多红色点片状沉积,N组茜素红染色为阴性;(4)DN组BMP-2和MGP呈显著负相关(r=-0.494,P<0.01),BMP-2表达与尿蛋白、血肌酐、血磷(r=0.422,P<0.01;r=0.743,P<0.01 r=0.671,P<0.01)呈显著正相关,MGP表达与尿蛋白、血肌酐、血磷(r=-0.511,P<0.01;r=-0.490,P<0.01;r=-0.420,P<0.05)呈显著负相关。结论:DN大鼠肾内小动脉钙化前已有血管钙化标志物BMP-2和MGP的表达,随时间延长BMP-2基因表达量逐渐增多,MGP基因表达量逐渐减少,提示DN大鼠肾脏内BMP-2及MGP基因表达量的变化可能参与DN进展。To observe the expressions of bone morphogenetic protein-2 （BMP-2） and matrix Gla protein （MGP） genes in kidney of diabetic nephropathy rats, and so as to explore their effects to diabetic nephropathy. Methods： Adult Sprague-Dawley male rats were divided into diabetic nephropathy group （group DN） and normal group （group N）. DN model was established by STZ induction. The expressions of BMP-2 and matrix Gla protein MGP gene were detected with Real-Time PCR in 4, 12 and 24 weeks. Calcium depositing around the renal arteriole was observed by alizarin red staining. Results： （ 1 ） Contents of blood glucose and urine protein of group DN at all time points were much higher than those of group N （P 〈 0.05 ）. Serum creatinine and serum phosphorus of group DN were higher than those of group N since the 12^th week（P 〈0.05）. （2） BMP-2 mRNA was detectable in group DN in 4 weeks, and gradually increased with the time passing, reached peak at the 12^thweek. The expression level of MGP mRNA of group DN was evidently mcreasect m ~＋ weeks, sllgtmy decreased in 12 weeks, reached bottom at the 12th week; No significant difference was found among the expressions at different time points in group N; （3） A little calcium salt was found deposited in the renal arteriole of DN rats in 4 weeks, and a large amount of calcium salt was found in 24 weeks. No calcium was found in control rats. （4）BMP-2 gene expression was negatively correlated with MGP gene expression in DN group （ r = - O.494, P 〈 0.01 ）. BMP-2 gene expression was positively correlated with serum creatinine, serum phosphorus and urine protein （r = 0. 422, P 〈 0. 01 ; r = 0. 743, P 〈 0.01 ;r =0. 671 ,P 〈0.01 ）. MGP gene expression was negatively correlated with serum creatinine, se- rum phosphorus and urine protein （r= -0.511, P〈0.0I; r= -0.490, P〈0.01; r = -0.420, P 〈0.05）. Conclusions： （1） BMP-2 and MGP mBNA occur in renal arteriole before the formation of vascular calcification. The expression

关 键 词：糖尿病肾病 骨形态发生蛋白质类 基质GLA蛋白 聚合酶链反应 大鼠 SPRAGUE-DAWLEY 

分 类 号：R34-33[医药卫生—基础医学] R587.1