摘要
目的基于蛋白质组学技术筛选与脾阳虚证相关的差异蛋白质,为进一步从分子水平上探讨脾阳虚证的本质提供有力的实验依据。方法 SPF级SD大鼠36只,随机分为正常对照组(16只)和脾阳虚证组(20只)。采用饮食不节、劳倦过度及苦寒伤阳药联合应用的方法,塑造脾阳虚证模型大鼠。分离、提取脾阳虚证组与正常对照组大鼠回肠的总蛋白,行双向凝胶电泳(two-dimensional electrophoresis,2-DE)技术,应用DelyderTM2D6.5图像分析软件识别差异蛋白点,应用MALDI串联飞行时间质谱(MALDI-TOF/TOF)技术获得相应肽质量指纹图,Mascot搜库鉴定差异蛋白质。结果经过统计学和模糊数学判定,脾阳虚大鼠模型造模成功。蛋白质组学初步鉴定,得到涉及细胞骨架、能量代谢及信号传导等多方面的差异表达的蛋白质8个,其中4个表达上调的蛋白分别是:结蛋白(desmin)、角蛋白8(cytokeratin 8,CK8)、丙酮酸激酶(pyruvate kinase,PK)、埃兹蛋白(ezrin);4个表达下调的蛋白分别是:甘油醛三磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase,GAPDH)、角蛋白19(cytokeratin 19,CK19)、角蛋白1(cytokeratin 1,CK1)、肌动蛋白(actin)。结论脾阳虚证状态下机体能量代谢速率减慢,能量生成减少,回肠绒毛蛋白结构变化,吸收消化功能减弱,这可能是脾阳虚证的病理机制。
Objective Based on proteomics technology, Pi-yang deficiency syndrome （PYDS） correlated differential proteins were screened, thus providing powerful experiment reliance for exploring the essence of PYDS. Methods Totally 36 SD rats of SPF grade were randomly divided into the normal control group （ n =16）and the PYDS group （ n =20）. The PYDS model rats were induced by improper diet, overstrain, and administration of yang impairing bitter cold herbs. The total proteins of the ileum were separated and extracted from rats in the PYDS group and the normal control group. The differential protein dots were identified using DelyderTM 2D 6.5 image analysis software by two-dimensional gel electrophoresis （2-DE）technology. The finger print map of corresponding peptide qualities was obtained by applying MALDI TOF/TOF. The differential proteins were identified using Mascot search library. Results Judged by statistics and fuzzy mathematics, Pi-yang deficiency rat model was successfully established. Eight proteins with differential expressions involving cell skeleton, energy metabolism, and signal transduction, and so on were obtained. Of them, there were 4 up-regulated proteins, i.e., desmin, cytokeratin8 （CK8）, pyruvate kinase （PK）, and ezrin. Four down-regulated proteins were glyceraldehyde-3-phosphate dehydrogenase （ GAPDH ）, cytokeratin19 （ CK19 ）, cytokeratinl （ CK1 ）, and actin. Conclusion The pathogenesis of PYDS might be slowed energy metabolism rate, reduced energy production, changed structure of ileal villin, and weakened absorbing and digestive functions.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2013年第1期71-75,共5页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.30973689)
关键词
脾阳虚证
比较蛋白质组学
回肠
Pi-yang deficiency syndrome
comparative proteomics
the ileum
