衰老大鼠主动脉增龄性改变的比较蛋白质组学研究  

Comparative proteomics analysis of aging rat aorta during the process of increasing age

作　　者：生玉平[1] 王琰[1] 赵韶华[1] 季翔[1] 高海青[1] 邱洁[1] 

机构地区：[1]山东大学齐鲁医院老年病科山东省心血管疾病蛋白质组学重点实验室,济南250012

出　　处：《中华老年医学杂志》2013年第1期91-95,共5页Chinese Journal of Geriatrics

基　　金：山东省优秀中青年科学家科研奖励基金（BS2009SW015）;山东省科技发展计划项目（2012GB021817）;山东省科技攻关计划项目（2010GGC10294）;国家科技部重大科技攻关项目分课题（20122×09303016-003）

摘　　要：目的研究老年大鼠主动脉与衰老相关的蛋白质，为进一步探讨衰老机制提供依据。方法建立衰老大鼠模型，取成年（9周）和老年（12个月）wistar_Kyoto（WKY）大鼠和同龄的自发性高血压大鼠（SHR），观察血压变化情况，采用苏木伊红（HE）染色观察主动脉形态学改变，应用同位素标记相对和绝对定量（iTRAQ）多重化学标记串联质谱技术鉴定差异蛋白，同时应用实时定量聚合酶链反应（PCR）、免疫印迹（Western-blot）检测部分差异蛋白。结果老年组中SHR大鼠的平均收缩压比WKY大鼠高出97．1％（t=39．00，P〈0．05）；成年组中SHR大鼠的平均收缩压比WKY大鼠高出5．40（t=3．64，P〈0．05）。HE染色结果显示老年WKY组和老年SHR组大鼠主动脉出现衰老改变，且老年SHR组更明显。质谱分析鉴定了7个与衰老相关的差异蛋白，分别是前纤维蛋白1（Profilin—1）、前层蛋白A（Prelamin—A）、热休克蛋白70（HSP70）、肌酸激酶M（CK—M）、腓骨蛋白5（Fibulin-5）、真核翻译起始因子5A_1（eIF5A）、抑制素蛋白（Prohibitin）。实时定量PCR结果显示，与成年组相比，WKY与SHR老年组中Prelamin—A的mRNA表达均升高，分别为0．45±0．04与0．15±0．01、0．78±0．06与0．34±0．02（t值分别为12．67、12．06，均P〈0．01）。与成年组相比，WKY与SHR老年组中Prohibitin mRNA表达降低，分别为1．34±0．05与1．01±0．06、1．24±0．05与0．88±0．08（t值分别为7．41、7．09，均P〈0．01）。Westernblot检测结果显示，与成年组相比，WKY与SHR老年组中Profilin-1的蛋白表达均升高，分别为9．12±0．40与20．76±0．80、16．84±0．50与55．16±0．90（t值分别为22．55、64．46，P〈0．01），且Profilin-1在SHR老年组的蛋白表达比WKY老年组的高，分别为55．16±0．90与20．76±0．80（t=49．49，P〈0．01）。结论通过比较蛋白质组学方法鉴定老�Objective To study the proteins related to aging in aortic of old rats for laying the foundation of further study of aging mechanism. Methods The rat model of aging was built,and all model rats were divided into 4 groups： the adult group（9 weeks）, the old group（12 months）of Wistar- Kyoto （WKY） rats, and the age-matched spontaneously hypertensive rats （SHR）. Blood pressure of 4 groups was observed. Morphological change of aorta was observed by HE staining. Differential proteins were identified by isobaric tags for relative and absolute quantitation, （iTRAQ）-coupled liquid chromatography and tandem mass spectrometry technology. Part of differential proteins was subsequently detected by real time PCR and Western blot. Results The mean SBP of the old group SHR was higher than WKY of 97.1%（t= 39.00,P〈0.05）, and the adult group of SHR was also higher than WKY of 5.4%（t=3.64,P〈0.05）. Compared with the adult group, aging change in the aortic morphology of old SHR and WKY were shown in HE staining, and the change in SHR rats was more marked. 7 proteins related to aging were identified by Mass spectrum analysis, and they were Profilin-1, Prelamin A, HSP70, creatine kinase-M, Fibulin-5, eIF5A and Prohibitin. Part of differential proteins was subsequently confirmed by real time PCR and Western blot. Prelamin-A was up-regulated in the old group of WKY and SHR （0. 15±0.01 vs.0. 45±0.04, 0.34±0.02 vs. 0. 78± 0.06） （t= 12.67,12.06, all P〈0.01）, Prohibitin was down-regulated in the old group of WKY and SHR（1.34±0.05 vs. 1.01±0.06, 1.24±0.05 vs. 0.88±0.08） （t=7.41, 7.09,all P〈0.01）. Profilin-1 was up-regulated in the old group of WKY and SHR （9.12±0.4 vs. 20.76±0.8, 16.84± 0. 5 vs. 55. 16±0.9） 0=22.55, 64.46,both P〈0.01）, and Profilin-1 expression in the old group of SHR was higher thanWKY （55.16±0.90 vs. 20.76±0.8, t=49.49,P〈0.01）. Conclusions Differential proteins of the old rat aorta are identified through the comparative proteomics method. These

关 键 词：衰老 主动脉 蛋白质组 

分 类 号：R285.5[医药卫生—中药学]