肥胖者脂肪组织基因表达谱特征及人类肥胖相关新基因C10orf116的功能研究

【作者】 倪毓辉；

【导师】 陈荣华； 郭锡熔；

【作者基本信息】 南京医科大学 ， 儿科学， 2006， 博士

【摘要】 肥胖是一种常见的能量代谢性疾病，是多种疾病(如：糖尿病、心脑血管疾病)的始发因素，在全球范围内已呈流行态势。近年来，肥胖低龄化的趋势日趋突出，儿童与青少年肥胖及并发症问题也引起广泛关注，因此积极探讨肥胖及其并发症的病因与发病机制已成为儿科学者目前重要的研究方向之一。 遗传因素在肥胖发生中的重要性已获得广泛认同。20世纪90年代以来，研究者从脂肪组织中克隆获得的多个肥胖相关基因，已为肥胖病因与发病机制的研究开拓了思路。然而，以这些基因为靶标开展的肥胖防治研究并未获得满意的临床疗效。因此，进一步克隆肥胖相关基因，并从基因网络角度分析这些基因在肥胖发生发展过程中的作用，可望为肥胖病因与发病机制的研究提供新的理论依据，为肥胖及其并发症的有效防治提供新的靶标。 本课题以抑制性差减杂交(suppression subtractive hybridization，SSH)技术，筛选肥胖与正常人腹膜后脂肪组织中的差异表达基因(differentially expressed genes)，以期分析肥胖者脂肪组织的基因表达谱特征，积极寻找肥胖相关新基因，并深入探讨其功能。 研究第一部分共筛查获得426个差异表达基因，其中216个特异高表达于肥胖者脂肪组织，210个特异高表达于正常人脂肪组织。进一步随机选取部分差异表达基因，应用RT-PCR技术验证其在肥胖与正常人脂肪组织中的表达趋势，结果发现其表达趋势与SSH结果一致，提示本课题SSH研究结果具有较高的可重复性。 生物信息学(Bioinformatics)分析发现，筛选获得的差异表达基因具有如下特征：(1)426个差异表达基因中，232个在生物数更多还原

【Abstract】 With the aim of identifying novel molecular pathways in human obesity, suppression subtractive hybridization (SSH) was performed in our study.Non-obese subjects and obese patients without any other diseases, such as endocrine diseases, metabolism diseases, cardiovascular diseases, malignant diseases and chronic inflammation, were recruited and the differentially expressed genes in omental adipose tissue were screened. Insulin resistance, as the common complication of obesity, was analyzed and excluded more strictly. Since the two groups of subjects have comparable state except weight and BMI, any differences in gene expression between groups are most likely due to obesity. In our study 426 genes were screened as the potential obesity-related genes, 216 obese up-regulated gene and 210 normal up-regulated gene. Several obesity related genes, such as ACRP30, Glut4, IGF1, LPL and aP2, were also screened in this study and in so doing served as compelling controls for the SSH procedure. To further verify and validate the SSH results, we performed RT-PCR analysis of 10 randomly chosen differentially expressed genes. Overall, the expression patterns obtained by RT-PCR reflected the results obtained by SSH, demonstrating the low false positive rate associated with SSH. 232 genes were identical to sequences that had previously been identified and characterized, among which, 174 genes had functional research, while 58 had no functional research. Besides, 158 genes were identical to previously identified更多还原