mTOR和VEGF基因表达在糖尿病大鼠肾损伤中的相关性  

Correlation of mTOR and VEGF Gene with Nephropathy in Diabetic Rats

作　　者：陶文玉[1] 杨莹[1] 熊煜欣[1] 李奕平[1] 彭辉[1] 杨涵麟[1] 陈郊丽[1] 谢余澄[2] 冯倩[3] 

机构地区：[1]昆明医科大学第四附属医院内分泌科,云南昆明650021 [2]昆明医科大学第四附属医院病理科,云南昆明650021 [3]昆明医科大学第四附属医院检验科,云南昆明650021

出　　处：《昆明医科大学学报》2016年第5期22-27,共6页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(30960148;81260135);云南省应用基础研究基金资助项目(2013FZ193);云南省中青年学术技术带头人后备人才基金资助项目(2011CI045);云南省医学学科带头人基金资助项目(D201217)

摘　　要：目的观察糖尿病大鼠mTOR和VEGF基因表达与肾损伤间的相关性.方法糖尿病(DM)大鼠成模后12、16、20、24周,选取DM组分别为7、6、6、9只和非糖尿病组(NDM)组各5只,测定肾小球及肾小管病理改变程度半定量评分(HS)及肾小球基底膜(GBM)厚度;免疫组化(IHC)测定mTOR、VEGF、VEGFR2抗体在肾脏中的表达;实时定量PCR(RT-Q-PCR)荧光染料法检测基因在肾脏的mRNA表达,基因的标化值(SCt)定量PCR产物.结果光镜下HS在总的DM组及各病程DM组均高于同期NDM组,20、24周DM组明显升高,GBM厚度在总的DM组明显高于NDM组,P<0.01,24周DM组增厚更明显;VEGF、VEGFR2基因IHC评分在总的DM组及各病程DM组均高于NDM组,两者在总的DM组中正相关,均与光镜下HS正相关,P<0.05.VEGF、VEGFR2基因SCt值在总的DM组均明显高于NDM组,VEGF基因SCt值在DM组间12周表达最高,24周次之,较16、20周明显升高,P<0.01;VEGFR2基因SCt值在DM组间随病程延长逐渐降低,24周最低.VEGF基因SCt值与VEGFR2基因SCt值在总的DM组明显正相关,P<0.01,与GBM厚度在总的DM组正相关.mTOR基因与VEGF、VEGFR2基因三者的IHC评分在总的DM组中均互相正相关,均与光镜下HS正相关,P<0.05.结论光镜下HS及GBM厚度在DM大鼠中升高,反应肾损伤,VEGF、VEGFR2基因蛋白及mRNA在DM大鼠肾脏中表达,且随着DM病程增长表达协同增高;VEGF mRNA表达与GBM厚度正相关;mTOR基因可能与VEGF、VEGFR2基因蛋白表达在DM大鼠肾损伤有协同相关作用,但不排外不同病程、不同动物样本量表达结果及实验条件可能存在结果差异.Objective The purpose of this study was to observe the correlation of mTOR and VEGF gene with nephropathy indicators in diabetic rats. Methods Forty-eight male Sprague-Dawley（SD） rats were divided into diabetes mellitus group（DM=28） and control group（NDM=20）. Diabetic models were produced by injection of streptozotocin. In the courses of 12,16,20 and 24 weeks, the histology scores（HS） and glomerular basement membrane（GBM） thickness were collected. The protein and m RNA expressions of the gene of mTOR,VEGF and VEGFR2 were observed by immunohistochemistry and real-time quantitative polymerase chain reaction（RT-Q-PCR） by SYBR Green. And the standardized cycle of threshold（SCt） was regarded as the indicators of the m RNA expression. Results HS and GBM thickness were significantly higher in DM rats than those in NDM rats,especially in DM rats of the courses of 20 and 24 weeks（P〈0.01）. IHC scores of VEGF and VEGFR2 were higher in total DM rats and were positively correlated with each other. There were positive correlations between HS with VEGF and VEGFR2 in total DM rats（P〈0.05）. SCts of VEGF and VEGFR2 were significantly higher and were positively correlated with each other in total DM rats（P〈0.01）. SCt of VEGF and GBM thickness showed positive correlation in total DM rats. SCt of VEGF was highest in the course of 12 w DM rats. SCt of VEGFR2 gradually decreased following by the diabetic course, and was lowest in the course of 24 w. There were no significantly differences in IHC scores and SCt of mTOR between DM and NDM rats. But the IHC scores of mTOR, VEGF and VEGFR2 were positively correlated with each other and with HS in total DM rats（P〈0.05）. Conclusion HS and GBM thickness were higher in diabetic rats, especially in the course of 24 w, which could reflect the injury of nephropathy. The protein and m RNA of VEGF and VEGFR2 were high expressed in kidney of DM rats and increased with the increasing of diabetic course. The m RNA expression of VEGF was positively co

关 键 词：MTOR VEGF 糖尿病大鼠 肾损伤 

分 类 号：R587.1[医药卫生—内分泌]