肥胖与非肥胖者腹部皮下及内脏脂肪组织蛋白质组差异的四例研究

Preliminary study on the differences of proteome in abdominal subcutaneous and visceral adipose tissue of obese and non-obese subjects

目的 采用二维电泳和质谱分析体系研究肥胖与非肥胖者腹部皮下及内脏脂肪组织的蛋白质组的差异 ,探讨胰岛素抵抗的机制。方法 采用固相 pH梯度二维电泳分离肥胖与非肥胖者腹部皮下及内脏脂肪组织的总蛋白质 ,采用凝胶银染显色 ,通过扫描仪获取凝胶图像 ,PDquest软件分析图像找出差异蛋白质点 ,对部分差异点用基质辅助激光解析电离飞行时间质谱仪 (MALDI TOF MS)测定其胶内酶解后的肽质量指纹图谱 ,并查询Mascot蛋白质数据库。结果 8个差异蛋白质点被初步鉴定为能参与细胞信号传导、代谢过程以及生长分化的蛋白质 ,分别为 :奎亭羧基末端水解酶 L1、AnnexinⅡ、组织相容性抗原ⅠA链、单酰甘油脂肪酶、血清反应元件的锌指结构域结合蛋白、AlloalbuminVenezia、PRO 2 0 4 4和未知蛋白。结论 差异表达的蛋白质在一定程度上提示腹部皮下及内脏脂肪组织之间可能存在功能差异 。

Objective To compare the proteins expressed in the abdominal subcutaneous and visceral adipose tissue of the obese and non obese subjects by using the technology of proteomics, and to find out the differently expressed proteins which might play a role in insulin resistance. Methods Two obese and two non obese subjects were studied. One gram of abdominal subcutaneous adipose tissue and one gram of visceral adipose tissue were acquired from each individual during the procedure. Two dimensional electrophoresis were carried out after sample preparation and protein quantification. The gel (silver stain) images were analyzed by PDquest software to find out the differentially expressed spots. Two dimensional electrophoresis was carried out for a second time for one of the samples. The gels were stained with Coomassie Brilliant Blue R250. The spots that showed the same site as those found in the silver stained gels were sectioned from the gels for mass spectrometry analysis. Peptide mass fingerprinting of eight spots were achieved. The eight spots were identified by searching the Mascot protein database. Results Eight spots were demonstrated, and they were PRO 2044 (NO.1), ubiquitin carboxyl terminal hydrolase L1 (NO.2), annexin Ⅱ(NO.3), monoglyceride lipase (NO.4), chain A of class Ⅰ histocompatibility antigen (NO.5), alloalbumin Venezia (NO.6), serum response element zinc finger domain binding protein (NO.7) and an unknown protein (NO.8). Conclusion Differentially expressed proteins in the abdominal subcutaneous and visceral adipose tissue of the obese and non obese subjects are found by using the technology of proteomics. These proteins are involved in signal conduction, metabolic process, growth and differentiation of the cells, The relationship of these proteins and insulin resistance remains unclarified and requires further study.