益肾抗纤复方对HK-2细胞转分化过程作用的基因组学研究  

Genomics Research on Roles of Yishen Kangxian Compound in the TEMT Process of HK-2 Cells

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

机构地区：[1]江苏省老年医院中医科,南京210024 [2]江苏省中医院肾内科,南京210029

出　　处：《中国中西医结合杂志》2013年第8期1129-1134,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：江苏省中医药管理局资助项目(No.LB09067)

摘　　要：目的通过基因芯片研究中药益肾抗纤复方与苯那普利含药血清对人肾近端小管上皮细胞(HK-2细胞)向间质成肌纤维细胞转分化(renal tubular epithelial cells to mesenchymal myofibroblasts transdif-ferentiation,TEMT)过程中的影响。方法制备SD大鼠含药血清,观察含药血清对HK-2细胞在5ng/mL转化生长因子β1(transforming growth factor-beta1,TGF-β1)诱导下TEMT过程的阻抑作用。HK-2细胞随机分为空白对照组、模型组、苯那普利组和中药组。采用IlluminaBeadChip的人全基因组基因表达芯片(HumanHT-12v4),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,苯那普利组与模型组比较差异表达基因227个,其中上调表达基因118个,下降表达基因109个;中药组与模型组比较差异表达基因97个,其中上调表达基因69个,下降表达基因28个。通过基因本体论分析,益肾抗纤复方较苯那普利在细胞损伤、凋亡、生长、NF-κB、蛋白激酶、神经元及血管生长等生物过程中有更多调控作用。结论益肾抗纤复方与苯那普利对HK-2细胞TEMT过程均有阻抑作用,但益肾抗纤复方对TEMT过程早期的细胞应对损伤、凋亡、生长等有更多途径的参与。Objective To study effects of Yishen Kangxian Compound （YKC） and benazepril containing serums on HK-2 cells （human renal proximal tubule epithelial cells） in the process of renal tu- bular epithelial cells to mesenchymal myofibroblasts transdifferentiation （TEMT） by gene chip. Methods YKC and benazepril containing serums were prepared. Their inhibitory effects on HK-2 cells in the trans- forming growth factor-beta1 （TGF-131）-induced TEMT process were observed. HK-2 cells were randomly di- vided into four groups, i.e.,the blank control group, the model group, the benazepril group, and the YKC group. The gross RNAs were extracted and purified by taking advantage of the HumanHT-12 v4 of Illumin- aBeadChip. Differentially expressed genes were obtained after they were reversely transcribed to cDNA, incorporating biotin labeling probe, hybridized with GeneChip,picture signals of fluorescence in gene ar- ray scanned and compared with differential genes by computer analysis. Results Differentially ex- pressed genes were successfully identified by gene chip. Compared with the model group, there were 227 differentially expressed genes in the benazepril group, including 118 up-regulated genes and 109 down- regulated genes. Compared with the model group, there were 97 differentially expressed genes in the YKC group, including 69 up-regulated genes and 28 down-regulated genes. The Gene Ontology （GO） a-nalysis indicated that YKC was more actively involved in the regulatory process than benazepril in terms of cell damage, apoptosis, growth, NF-KB, protein kinase, neuron, and blood vessel growth. Conclu- sions YKC and benazepril could inhibit the TEMT process of HK-2 cells. But YKC also had taken part in cell damage, apoptosis, growth ,and more pathways of early stage TEMT.

关 键 词：益肾抗纤复方 苯那普利 人肾近端小管上皮细胞 肾小管上皮细胞向间质成肌纤维细胞转分化 

分 类 号：R285.5[医药卫生—中药学]