埃他卡林对重要器官心脑肝基因表达的影响  

Analysis of Iptakalim-induced gene expression changes in heart, brain and liver by cDNA microarray

作　　者：王玉[1] 潘志远[1] 汪海[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《中国药理学通报》2007年第3期302-310,共9页Chinese Pharmacological Bulletin

基　　金：国家高技术研究发展计划(863计划)重大专项基金资助项目(No2002AA2Z3137)

摘　　要：目的利用BioStarR40s cDNA芯片,检测经全新结构类型化合物埃他卡林3mg·kg-1灌胃2wk处理后的大鼠心脏、脑、肝脏组织基因表达的变化,初步判断埃他卡林的基因作用谱;同时探讨埃他卡林给药2wk后,其心脏血流动力学的变化、心肌形态学和超微结构的变化,为阐述埃他卡林的体内生物学作用特征和评价其长期用药安全性提供实验依据。方法①给予埃他卡林(3mg·kg-1)2wk后取大鼠心脏、脑、肝脏组织,抽提总RNA,反转录合成cDNA探针;将荧光标记的cDNA探针与BioStarR-40S芯片杂交,对荧光信号扫描分析。结合RT-PCR方法对芯片结果进行验证。②Wistar♂大鼠随机分为5组:正常对照组、埃他卡林1、3、9mg·kg-1组以及9mg·kg-1撤药组。采用直接插管法,观察麻醉大鼠的血压、心率、收缩参数以及舒张参数的变化;采用HE染色和电镜技术观察不同组对心肌的形态学和超微结构的变化。结果①埃他卡林对心、脑和肝脏基因表达的调节作用具有选择性。采用4096个具有重要功能的基因序列的芯片,埃他卡林3mg·kg-1灌胃2wk,大鼠心脏236个基因的表达发生变化,其中100个基因表达上调,136个基因表达下调;肝脏6个基因表达上调;脑组织的基因表达未发生变化;埃他卡林调节心脏组织的基因表达包括与肌肉收缩相关的肌球蛋白、肌动蛋白和微管蛋白等;与信号转导相关的G蛋白、腺苷酸环化酶和钙调结合蛋白等;②埃他卡林1、3、9mg·kg-1灌胃2wk,在体心脏功能未发现有药理学意义的变化,也未发现心肌组织及其超微结构有病理学意义的变化。结论该实验体系下埃他卡林对重要器官、心、脑和肝脏基因表达的调节作用具有选择性;反复给予埃他卡林未发现其对心脏的不良反应。Aim To observe changes of gene expression profile in rat heart , liver and brain treated with iptakalim using cDNA microarray and analyze its molecule mechanism about pharmacology and treatment. Methods ①Rats were treated with iptakalim for 14 days at a dose of 3 mg · kg^-1 by garage. Total RNA was extracted from heart, liver and brain tissues and reversely transcribed to eDNA, which were labeled with Cy3 and Cy5 fluorescent as probes and hybridized to Gene Chip （ BiostarR-40s ）. Scan array 3 000 was used for scanning the hybridizing signals and Image-GENE 3.0 for data analysis. Data were confirmed using RT-PCR. ②Rats were randomly divided into normal control group and 4 iptakalim groups. Iptakalim was administered orally at doses of 1, 3, 9 mg · kg^-1 body weight per day for 2 weeks. The last group was orally administered at a dose of 9 mg · kg^- 1 body weight per day for 2 weeks, and iptakalim was withdrawn for 10 days. We observe effects of iptakalim on hemodynamics parameters in anesthetized rats and changes of myocardial structure, myofilament ultra - structure after 24h at the end of the last administration. Results ① The new chemical entity iptakalim had selectivity on changes of gene expression in rat heart, liver and brain. Compared with control group, 236 genes are changed （100 increased and 136 decreased） in rat hearts and 6 transcripts （6 increased） in rat livers, there are no changes on gene expression in rat brains. ② In anesthetized rats, iptakalim at doses of 1, 3, 9 mg · kg^-1 neither affected pharmacological effects on cardiovascular hemodynamics parameters nor had pathological changes on myocardial morphological and ultrastructure. Conclusion Under the same experimental conditions, the new chemical entity iptakalim has selectivity on changes of gene expression in vital organs. Iptakalim shows no side-effects on cardiac function and tissue structure.

关 键 词：基因芯片 埃他卡林 基因表达 心功能 

分 类 号：R-332[医药卫生] R322.11