Gene expression in obese humans before and during treatment with rimonabant. Homo sapiens

Rimonabant was prescribed to eight patients, admitted to the Overweight Study Unit at the Department of Medicine, Karolinska University Hospital. All patients had a BMI > 35 kg/m2, were treated on clinical indications (metabolic and mechanical disability) and without mental disturbances. Blood samples were collected before treatment and at the first clinical control, when the patients had received rimonabant, 20 mg daily, for 4 weeks. Overall design: Blood mononuclear cells were isolated by Ficoll separation (Ficoll-Paque PLUS, GE Healthcare). From the cell-pellet total RNA was prepared using Qiagen midi plus kit (Qiagen GmbH, Hilden Germany) as recommended by the manufacturer and was quality controlled on an Agilent Bioanalyzer (Agilent Technologies, Inc. Palo Alto, CA). Six pretreatment samples and seven samples obtained after rimonabant treatment passed the quality control. The cRNA synthesis for microarray experiments and the hybridizations were carried out using Affymetrix Human Gene 1.0 ST Array (Affymetrix, Inc., Santa Clara, CA) according to standard Affymetrix protocols at the core facility for Bioinformatics and Expression Analysis, Department of Biosciences and Nutrition, Karolinska Institutet.本研究纳入瑞典卡罗林斯卡大学医院（Karolinska University Hospital）内科超重研究病房的8名受试者，所有受试者的身体质量指数（Body Mass Index, BMI）均大于35 kg/m²，且因代谢与机械性功能障碍的临床指征接受治疗，无精神类疾病史。分别于治疗前，以及受试者每日服用20mg利莫那班（Rimonabant）、持续4周后的首次临床随访时采集血液样本。 实验整体流程如下：采用菲科分离法分离外周血单个核细胞（Blood mononuclear cells），所用试剂为Ficoll-Paque PLUS（通用电气医疗，GE Healthcare）；依照制造商推荐操作指南，使用Qiagen midi plus试剂盒（Qiagen GmbH，德国希尔德）从细胞沉淀中提取总RNA，并通过安捷伦生物分析仪（Agilent Bioanalyzer，安捷伦科技公司，Agilent Technologies, Inc.，美国加州帕洛阿尔托）完成质量质控。最终共有6份治疗前样本与7份利莫那班治疗后样本通过质控检测。 在卡罗林斯卡学院（Karolinska Institutet）生物科学与营养系的生物信息学与表达分析核心平台，按照Affymetrix标准实验流程，使用Affymetrix人类基因1.0 ST芯片（Affymetrix, Inc.，美国加州圣克拉拉）完成微阵列实验所需的互补RNA（complementary RNA, cRNA）合成与杂交实验。