通过基因芯片研究苯那普利对HK-2细胞TEMT过程的影响作用  

Research on the impacts of benazepril on TEMT process in HK-2 cells through gene chips

作　　者：孙世竹[1] 陈燕[2] 鲍军[1] 秦雪梅[1] 蒋娟娟[2] 申志祥[2] 高雪燕[2] 

机构地区：[1]江苏省老年医院中医科,江苏省南京市210024 [2]江苏省老年医院肾科,江苏省南京市210024

出　　处：《实用老年医学》2012年第1期26-30,52,共6页Practical Geriatrics

基　　金：江苏省中医药管理局资助课题(LB09067)

摘　　要：目的通过基因芯片探讨苯那普利对人肾近端小管上皮细胞(HK-2细胞)向间质肌成纤维细胞转分化(TEMT)过程中的影响作用。方法制备苯那普利含药血清,将HK-2细胞随机分为正常组、模型组、苯那普利(洛丁新)组。采用Illumina Bead Chip的人全基因组基因表达芯片(Human HT-12 v4),提取总RNA分离纯化后,经过逆转录、合成、杂交、洗片和扫描获得差异表达基因,并进行差异基因相关分析。结果通过基因芯片技术成功筛选差异表达基因,模型组与正常组比较差异表达基因80个,其中上调表达53个,下降表达27个;洛丁新组与模型组比较差异表达基因227个,其中上调表达118个,下降表达109个。通过基因GO分析,洛丁新组较模型组在细胞增殖、移动、转分化、连接酶活化等生物过程中有更多参与。结论本实验为更加全面阐明苯那普利对TEMT的调控作用提供了实验依据。Objective To explore the mediating impacts of benazepril on epithelial-to-mesenchymal transition in human renal proximal tubule epithelial cells induced by TGF-131 （TEMT） through the gene chip. Methods To prepare benazepril drug-containing serum, HK-2 cells were randomly divided into normal group, model group and benazepril group. The differentially expressed genes were obtained by applying Human HT-12 v4 of Illumina Bead Chip after the total RNAs were isolated from the three groups by TRIzol-method and then purified, reversely transcribed to cDNA, incorporating biotin labeling probe, hybridizated with genechip, incubation of HK-2 cell with TGF-β1 and drug-containing serum at 48 h. Picture signals of fluorescence in gene array were scanned and compared with differential expression of genes by computer ana- lysis. Results Eighty differentially expressed genes were successfully identified in the model group compared with the normal group, including 53 upregulated and 27 downregulated genes;221 differentially expressed genes were identified in the benazepril group compared with the model group, including 118 upregulated and 109 downregulated genes. Based on gene ontology （GO） analysis, the benazepril group was more actively involved in biological process than that in model group in cell proliferation, movement, differentiation and ligase activation. Conclusions The research provides an experimental basis for a more comprehensive elaboration on the regulating role of benazepril in EMT.

关 键 词：苯那普利 人肾近端小管上皮细胞(HK-2细胞) 肾小管细胞上皮细胞向间质肌成纤维细胞转分 化(TEMT) 基因芯片 差异表达基因 基因GO分析 

分 类 号：R972.4[医药卫生—药品]