亚慢性砷暴露大鼠血清脂代谢相关差异表达蛋白的筛选和验证  

Screening and validation of differentially expressed serum proteins related to lipid metabolism in rats under subchronical arsenic exposure

作　　者：张锦慧[1] 宋选波 李颖[1] 纪晓红[1] 孙洪娜[1] 王慧 赵丽军[1] 孙殿军[3] 

机构地区：[1]哈尔滨医科大学中国疾病预防控制中心地氟病防治研究所国家卫生计生委病因流行病学重点实验室(23618504)、黑龙江省普通高校病因流行病学重点实验室,哈尔滨150081 [2]上海市嘉定区南翔镇社区卫生服务中心,上海201802 [3]哈尔滨医科大学中国疾病预防控制中心地方病控制中心,哈尔滨150081

出　　处：《中华地方病学杂志》2016年第6期401-405,共5页Chinese Journal of Endemiology

基　　金：国家自然科学基金（81202155）

摘　　要：目的筛选亚慢性砷暴露大鼠血清差异表达蛋白，并对与脂代谢有关的蛋白进行验证．为研究砷暴露人群脂代谢相关疾病的发生提供线索。方法将60只Wistar大鼠按体重采用随机数字表法分为对照组和低、中、高剂量染砷组，各组饮水中亚砷酸钠含量分别为0、2、10、50mg／L，每组15只。实验周期为12周。利用相对和绝对同位素定量标记（Isobaric tags for relative and absolute quantitation，iTRAQ）蛋白质组学技术筛选对照组与高剂量染砷组之间的血清差异表达蛋白。采用酶联免疫吸附试验（ELISA）法对筛选出的可能与脂代谢有关的差异表达蛋白进行验证。结果大鼠血清中共鉴定出201个蛋白，有12个蛋白的表达量在对照组和高剂量染砷组间存在显著性差异，其中与脂代谢有关的为载脂蛋白A-Ⅱ（上调：高剂量染砷组／对照组=1．40）和脂联素（下调：高剂量染砷组／对照组=0．75）。ELISA法验证结果表明，对照组和低、中、高剂量染砷组大鼠血清载脂蛋白A-Ⅱ含量分别为（4．37±1．08）、（5．04±0．37）、（4．94±0．84）、（4．65±0．61）μg／L，组间比较差异无统计学意义（F=2．071，P〉0．05）；4组大鼠血清脂联素含量分别为（1．06±0．43）、（0．95±0．42）、（0．88±0．28）、（0．54±0．17）mg／L，与对照组比较，高剂量染砷组表达水平显著下降（P〈0．05）。结论利用iTRAQ蛋白质组学技术筛选出一组与亚慢性砷暴露有关的大鼠血清差异表达蛋白，并初步验证脂联素有可能作为砷暴露致健康损害的生物标志物。Objective To screen and validate the differentially expressed serum proteins related to lipid metabolism in rats under subchronical arsenic exposure, and to provide serum protein biomarkers for lipid metabolism related diseases. Methods Sixty Wistar rats were randomly divided into four groups, and the dose of sodium arsenite in drinking water for each group was 0, 2, 10, and 50 mg/L, respectively, n = 15/group. The exposure lasted for 12 weeks. An isobaric tags for relative and absolute quantitation （iTRAQ）-based proteomic technique was used to identify the differential expressed proteins in serum between control and high-arsenic groups. Enzyme-linked immunosorbent assay （ELISA） method was applied to further validate differentially expressed proteins potentially related to lipid metabolism in the four groups. Results Totally 201 proteins were identified in rat serum, of which 12 were significantly changed which related to lipid metabolism including apolipoprotein A- Ⅱ （up-regulated, high-arsenic group/control = 1.40） and adiponectin （down-regulated, high-arsenic group/control = 0.75）. Validation of ELISA experiment showed that the concentrations of serum apolipoprotein A-II in control, lowarsenic, middle-arsenic, and high-arsenic groups were （4.37 ±1.08）, （5.04 ± 0.37）, （4.94 ± 0.84） and （4.65±0.61） μg/L, respectively, and no significant difference was found among the four groups （F = 2.071, P 〉 0.05）. The concentrations of serum adiponectin were （1.06 ±0.43）, （0.95±0.42）, （0.88 ± 0.28） and （0.54±0.17） mg/L, and the value in high-arsenic group was significantly decreased as compared to that of control （P 〈 0.05）. Conclusion This study has demonstrated that iTRAQ technology can be used in differential expression proteomics, and adiponectin may serve as a useful biomarker for damage to human health caused by arsenic poisoning.

关 键 词：砷暴露 相对和绝对同位素定量标记 血清 生物标志物 载脂蛋白A-Ⅱ 脂联 素 

分 类 号：R114[医药卫生—卫生毒理学]