【作者】 薛峰；

【导师】 江华； 林子豪； 吴宏； 王文静； 袁相斌； 赵耀忠；

【作者基本信息】 第二军医大学 ， 外科学， 2006， 硕士

【摘要】 背景：病理性瘢痕在临床上可分为增生性瘢痕和瘢痕疙瘩两种类型。二者均由胶原纤维和细胞以及新生血管构成，其特征是大量的成纤维细胞增生和过度的胶原沉积。成纤维细胞是产生胶原的主要细胞，与病理性瘢痕的形成有重要关系。随着现代细胞生物学和分子生物学的迅猛发展，有人利用基因芯片技术对瘢痕组织与正常皮肤进行研究，已经证实了基因表达差异的存在。这些研究结果为本课题提供了有力的基因学依据。蛋白质芯片指纹图谱(SELDI)技术将蛋白质芯片与飞行质谱相结合，使它既具有芯片的高通量、高效率和重复性好的特点，又具有飞行质谱的高灵敏度检测的功能，可直接对血清及组织粗样品中的各种蛋白质进行检测，其理论和技术的发展为各种疾病的研究提供了新的方法。 目的：对体外培养的增生性瘢痕及正常皮肤组织中的成纤维细胞，利用赛弗吉的SELDI-TOF-MS(表面加强激光解吸电离飞行时间质谱)技术，分别建立增生性瘢痕成纤维细胞与正常皮肤组织成纤维细胞的蛋白质指纹图谱，找寻两者的蛋白质表达差异，建立两者的蛋白质指纹差异图谱，为增生性瘢痕形成机制的研究提供新的思路。 方法：选择增殖期的增生性瘢痕18例，部分瘢痕个体的正常皮肤组织6例。标本全部手术切取，立即行细胞培养。经3～4次传代后，将成纤维细胞裂解，裂解液上样于蛋白质芯片，用SELDI-TOF-MS芯片阅读仪读取数据，建立增生性瘢痕及正常皮肤组织成纤维细胞的蛋白质指纹图谱。进一步用生物信息学方法分析比较所得数据，选择有意义的蛋白质位点，建立一个敏感可靠的的生物信息学模型。 结果：共检测出394个波峰，其中m／z值在2～5KD之间的有143个；5～10KD之间的71个；10～20KD之间的47个；20～30KD之间的18个；30～40KD之间的34个；40～50KD之间的17个。主要的峰值集中在2～10KD之间。经过Ciphergen Biomarker Wizard软件处理，得出m／z位于2805Da、2910Da、更多还原

【Abstract】 Background: Pathological scars may be divided into two kinds: hypertrophic scars and keloids. They are all made up of collagen fibers, cells and new-born blood vessels with the characteristic of fibroblasts’ hyperplasia and collagen’ s over-deposit. Whereas the collagen is secreted mainly by fibroblasts, which, it is concerned , has a close relationship with the formation of pathological scars. Along with the rapid development of cellular and molecular biology in recent years, the comparison of gene profiling of normal skin with hypertrophic scars has identified some gene segment responsible for the formation of scar. Such are important evidences for our study. SELDI is a new technology which developed after gene chip. SELDI combines protein chip technology with Mass Spectrometry . It has some merits such as high quantum, high efficiency, good repeatability of chip , and high sensitivity of Mass also. The development of knowledge and technology in proteomics has provided a powerful method in studying diseases.Objective: To screen and evaluate protein biomarkers for the fibroblastic cells of hypertrophic scar and normal skin. By Ciphergen’s SELDI-TOF-MS , all different protein spots were detected from the protein chips. From the point set, some special spots were picked out as the part of desired protein fingerprint.. Combining with bioinformatics, a specific patterns was established successfully to discriminate hypertrophic scar from normal skin.Methods: The fibroblastie cells from eighteen cases of hypertrophic scar and six cases of normal skin were cultured individually in vitro.更多还原