摘要
目的基于血清差异蛋白质组学技术研究强迫游泳应激抑制大鼠自然杀伤细胞（natural killer cells，NK）杀伤活性的机制。方法采用游泳力竭法制备应激大鼠模型，大鼠随机分为正常对照（A）组，强迫游泳（B）组，每组12只。MTT法检测NK细胞杀伤活性，酶联免疫吸附方法（enzyme linked immunosorbent assay，ELISA）检测血清γ-干扰素（interferon-γ，INF-γ）水平，采用iTRAQ蛋白质组学技术筛选差异表达蛋白，并采用ELISA方法验证蛋白组学检测结果的可靠性。结果与正常对照组大鼠相比，强迫游泳应激组大鼠NK细胞杀伤活性和血清IFN-γ水平降低（t＝3.68和t＝2.76，P〈0.05）。蛋白质组学检测结果表明，与正常对照组大鼠相比，强迫游泳应激大鼠111个蛋白质表达发生明显变化，其中47个蛋白质表达明显增加，54个蛋白质表达明显下降。ELISA验证结果与蛋白质组学检测结果一致。结论蛋白质组学筛选的差异表达蛋白对探讨慢性应激致免疫功能降低的机制有重要意义。
ObjectiveTo explore mechanisms of suppressed cytotoxicity of natural killer（NK） cells induced by forced swimming stress in Rats based on proteomics technology.MethodsThe forced-swimming rat model was made.The rats were divided into 2 groups： normal control group （A） and forced swimming group （B）, with 12 rats in each group. MTT assay and enzyme linked immunosorbent assay （ELISA）were used to detect cytotoxicity of NK cells and serum levels of IFN-γ. In addition, iTRAQ and ELISA were also adopted to screen different proteins expressed and to verify the reliability of results of proteomics analysis.ResultsCompared to normal control rats, the cytotoxicity of NK cells and serum levels of IFN-γ in forced swimming rats were both decreased（t=3.68 and t=2.76, both P〈0.05）. Proteomics analysis showed that compared to normal control rats, there were 111 differentially expressed proteins in forced swimming rats, including 47 proteins up-regulated, 54 proteins down-regulated.The result of ELISA results were consistent with the results of proteomics analysis.ConclusionIt is improtant to use proteomics to explore the mechanisms of chronic stress induced disorder.
作者
侯殿东
王德成
高嵋
马贤德
雷萍
徐铭
韩晓伟
关洪全
Hou Diandong;Wang Decheng;Gao Mei;Ma Xiande;Lei Ping;Xu Ming;Han Xiaowei;Guan Hongquan(Department of Pathology Immunology,Basic Medical College,Liaoning University of Traditional Chinese Medi-cine,Shenyang 110847,China;Biotechnology Laboratory Experiment and Technology Center,Department of Clinical Laboratory,Second Affiliated Hospital,Liaoning Univer-sity of Traditional Chinese Medicine,Shenyang 110034,China,Electric Inspection Section,Affiliated Hospital,Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China)
出处
《国际免疫学杂志》
CAS
2018年第3期260-264,共5页
International Journal of Immunology
基金
国家自然科学基金（81303079）
辽宁省高等学校杰出青年学者成长计划（LJQ2015072）
关键词
强迫游泳应激
蛋白质组学
自然杀伤细胞
Comparative proteomics analysis
Forced swimming stress
Natural killer cells
