【作者】 顾水明； 魏盟； 张昀昀； 邵君； 李奇； 吴宗贵；

【Author】 GU Shui-ming~(1),WEI Meng~(1),ZHANG Yun-yun~(1),SHAO Jun ~(3),LI Qi ~(3),WU Zong-gui ~(2)(1.Department of Cardiology,Affiliated No 6 Hospital,Shanghai Jiaotong University,Shanghai,200233;2.Department of Cardiology,Changzheng Hospital,Second Military Medical University,Shanghai 200003,China;3.Shanghai National Engineering Center for Biochip,Shanghai 201203,China)

【机构】 上海交通大学附属第六人民医院心内科； 生物芯片上海国家工程研究中心； 第二军医大学附属长征医院心内科；

【摘要】 目的:应用基因芯片技术分析比较慢性心力衰竭(CHF)患者和健康对照组外周血单核细胞(PBMC)中细胞因子和其它炎症相关基因表达差异,以识别CHF细胞因子网络的不平衡。方法:分别抽提CHF和健康对照组PBMC总RNA并纯化mRNA,反转录合成单链、双链cDNA后,体外转录合成cRNA。分别用cy3-dCTP和cy5-dCTP标记对照和CHF组cRNA。将基因芯片和杂交探针变性后杂交、洗涤。用基因芯片扫描仪进行图像扫描,Im aGene3.0软件分析cy3、cy5两种荧光信号的强度和比值。结果:cDNA表达谱芯片中含365个炎性细胞因子相关基因,其中CHF组上调34个基因,下调2个。上调基因包括肿瘤坏死因子(TNF)超家族成员,趋化因子/趋化因子受体,转化生长因子超家族成员等。TNF超家族成员中,淋巴毒素(LTB)、TNF超家族成员5、7、10,TNF受体超家族成员13b,TNF相关凋亡诱导配体受体4(TRAIL-R4)在CHF组均上调。而TNF受体超家族成员10C(TRAIL-R3)在CHF显著下调。结论:CHF与正常人PBMC细胞因子相关基因的表达有显著差异,CHF存在细胞因子网络的不平衡;尤其是多个TNF超家族基因的高表达可能在CHF发生发展中起着重要作用。更多还原

【Abstract】 AIM: To investigate the gene expression difference of cytokines and related mediators in(peripheral) blood mononuclear cells(PBMC) between chronic heart failure(CHF) patients and healthy subjects,and identify the imbalance in the cytotokine network in CHF.METHODS: Total RNA was extracted from PBMCs of CHF patients and healthy subjects and transcripted to single,two stranded cDNA and cRNA.Two probes were labled with cy3 and cy5 fluorescence,mixed and hybridized with the gene chip.The hybridizing signals were deetected by laser scanner and analysed with and ImaGene 3.0 software.RESULTS: Among the 365 cytokines and related mediator genes in the gene chip,34 were upregulated and 2 downregulated in CHF patients.The regulated genes included several members of tumor necrosis factor(TNF) superfamily,chemokines/-receptors,members of the transforming growth factor superfamily.Of the TNF superfamily members,lymphotoxin,TNFSF5,TNFSF7,TNFSF10,TNFRSF13B,and TRAIL-R4 were upregulated,while TNFRSF10C(TRAIL-R3) was downregulated.(CONCLUSION): There are significant differences in gene expressions of cytokines and related mediators in PBMCs between CHF patients and healthy subjects and the imbalance in the cytokine network in CHF.In particular, the enhanced expression of several members in the TNF superfamily may play a potential pathogenic role in CHF.更多还原