桂枝加葛根含药血清干预纤维环细胞的蛋白组学差异  

Proteomic difference of serum containing Cinnamon Twig Decoction Plus Pueraria on annulus fibrosus cells

作　　者：仲卫红[1] 李宇涛[2] 郑其开[1] 叶佳佳[1] 林建平[1] 王诗忠 

机构地区：[1]福建中医药大学附属康复医院,国家中医药管理局中医康复技术三级实验室,福建中医药大学省级中医药重点研究室,脊柱与骨关节运动功能康复研究室,福建省康复技术工程研究中心,福建省福州市350003 [2]福建中医药大学,福建省福州市350003 [3]福建生物工程职业技术学院,福建省福州市350003

出　　处：《中国组织工程研究》2014年第11期1718-1723,共6页Chinese Journal of Tissue Engineering Research

基　　金：福建省自然科学基金资助项目(2010J01183)~~

摘　　要：背景:研究表明纤维环破裂是颈椎病椎间盘退变的诱发因素。课题组既往研究表明桂枝加葛根汤治疗颈椎病的疗效是肯定的,但有关桂枝加葛根汤干预纤维环细胞的差异蛋白组学研究尚未见报道。目的:从差异蛋白组学的角度,分析桂枝加葛根汤对椎间盘纤维环细胞作用的靶位蛋白或相关蛋白。方法:体外培养SD大鼠纤维环细胞,将传3代纤维环细胞随机分为桂枝加葛根汤组和空白组,分别加入含自制桂枝加葛根汤血清的培养液以及含生理盐水血清的培养液干预。运用同位素标记相对和绝对定量技术标记2组样本的蛋白,二维液相色谱质谱仪(2D LC-MS/MS)鉴定蛋白并进行相对定量分析。结果与结论:以空白组作为对照,在桂枝加葛根组干预纤维环细胞后血小板衍生生长因子A、骨形态发生蛋白2、胰岛素样生长因子1、聚集蛋白聚糖核心蛋白、转化生长因子β3、白细胞介素1β蛋白表达上调;同时碱性成纤维生长因子、Ⅰ型胶原α1链蛋白表达下调。提示桂枝加葛根汤对纤维环细胞的退变有修复作用。BACKGROUND：Anulus fibrosus cleavage is the main contributing factor of intervertebral disk degeneration in cervical spondylosis. Previous studies of our research group have demonstrated that, Cinnamon Twig Decoction Plus Pueraria is a definitely effective treatment of cervical spondylosis. However, differential proteomic research addressing the intervention of Cinnamon Twig Decoction Plus Pueraria on anulus fibrosus cel s is rarely reported. OBJECTIVE：To explore the target or related proteins of Cinnamon Twig Decoction Plus Pueraria intervention on anulus fibrosus cel s, at the differential proteomics level. 〈br〉 METHODS：Anulus fibrosus cel s were col ected from Sprague-Dawley rats that were cultured in vitro. Cultivated anulus fibrosus cel s at passage 3 were randomly divided into two groups：experiment group was cultured in serum containing Cinnamon Twig Decoction Plus Pueraria, while control group in saline. The isotopical y labeled relative and absolute quantification was applied to mark the sample proteins in two groups. Two-dimensional liquid chromatography mass spectrometry was used to identify the proteins and relative quantitative analysis was performed. RESULTS AND CONCLUSION：Compared with the control group, platelet-derived growth factor subunit A, bone morphogenetic protein 2, insulin-like growth factor I, aggrecan core protein, transforming growth factor beta-3, interleukin-1 beta expression were increased in the experiment group, while fibroblast growth factor 2 and col agen alpha-1 chain expression was decreased. Cinnamon Twig Decoction Plus Pueraria can repair the degeneration of anulus fibrosus cells.

关 键 词：组织构建 组织工程 纤维环细胞 颈椎 桂枝加葛根汤 含药血清 二维液相色谱质谱仪 差异蛋白质组学 福建省自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]