中国中药杂志

2020, v.45(19) 4719-4724

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红景天苷干预前后高原红细胞增多症大鼠红细胞膜的蛋白质组学研究
Proteomics analysis of erythrocyte membrane in rats with high altitude polycythemia before and after intervention with salidroside

贾守宁;张庆光;马春花;李欣;王双玺;陈文娟;窦萱;葛广波;
JIA Shou-ning;ZHANG Qing-guang;MA Chun-hua;LI Xin;WANG Shuang-xi;CHEN Wen-juan;DOU Xuan;GE Guang-bo;Qinghai Hospital of Traditional Chinese Medicine;Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine;

摘要(Abstract):

研究红景天苷对高原红细胞增多症(high altitude polycythemia, HAPC)大鼠红细胞膜的蛋白质组学影响,基于蛋白质组学分析结果探讨红景天苷改善HAPC的机制。首先建立HPAC大鼠模型,16只大鼠造模成功后随机分为HAPC模型组和红景天苷100 mg·kg~(-1)组,每组8只。HAPC模型组灌胃生理盐水,红景天苷组大鼠灌胃给予100 mg·kg~(-1)的红景天苷,每日1次,连续灌胃40 d后股动脉采血,提取红细胞膜总蛋白,采用双向电泳将总蛋白进行分离;对红景天苷干预前后红细胞膜总蛋白双向电泳图谱进行差异分析,通过质谱技术鉴定具有明显差异的蛋白质,最后利用生物信息学分析生物学功能。结果显示,应用双向电泳法建立了HAPC分辨率较高且重复性较好的大鼠红细胞膜蛋白表达图谱。红景天苷干预红细胞膜双向凝胶电泳(two-dimensional gel electrophoresis, 2-DE)图谱中18个蛋白点发生明显变化,其中13个表达上调,5个表达下调;并借助质谱分析成功鉴定了其中8个差异性蛋白。进一步生物信息分析发现这些差异性蛋白参与了氧化应激、氧化还原、过氧化物酶体途径等生物过程,且主要与过氧化物酶体途径和丝裂原活化蛋白激酶(mitogen-activated protein kinase, MAPK)信号通路有关。因此,红景天苷可使HAPC大鼠红细胞膜蛋白表达发生较为明显的改变,借助蛋白质组学技术鉴定了其中8个差异蛋白,发现这些差异性蛋白参与氧化应激、氧化还原、过氧化物酶体途径等生物过程。
To investigate the effect of salidroside on the proteomics of erythrocyte membrane in high altitude erythrocytosis(HAPC) rats, in order to explore the mechanism of salidroside in improving HAPC based on the proteomics analysis. First, HPAC rat models were established, and 16 rats were randomly divided into HAPC model group and salidroside(100 mg·kg~(-1)) treatment group(8 rats per group). Saline was administered to the HAPC model group, while salidroside treatment group was given 100 mg·kg~(-1) salidroside once a day. After continuous oral administration with salidroside for 40 days(once a day), blood was collected from the femoral artery to obtain total red blood cell membrane proteins. Two-dimensional electrophoresis was used to separate total proteins. The two-dimensional electrophoresis of erythrocyte membrane proteins was analyzed before and after salidroside intervention, and the proteins with significant differences were identified by mass spectrometry. Finally, biological functions were analyzed using bioinformatics. A two-dimensional electrophoresis method was used to establish a protein expression profile with a high resolution and reproducibility of erythrocyte membranes in HAPC rats. Salidroside treatment significantly changed 18 protein spots in the 2-DE map of erythrocyte membranes, of which 13 proteins were up-regulated and 5 proteins were down-regulated. Eight differential proteins were successfully identified by mass spectrometry. Moreover, bioinformatics analysis found that these differential proteins were involved in such biological processes as oxidative stress, redox, and peroxisome pathway, which are mainly associated with peroxisome and MAPK signaling pathways. Therefore, salidroside could significantly change the expressions of erythrocyte membrane proteins in HAPC rats. Eight differential proteins were identified by a proteomic-based approach. The differential proteins were involved in such biological processes as oxidative stress, redox, peroxisome pathway.

关键词(KeyWords): 红景天苷;高原红细胞增多症;红细胞膜;蛋白质组学
salidroside;high altitude polycythemia;erythrocyte membrane;proteomics

Abstract:

Keywords:

基金项目(Foundation): 青海省科学技术厅应用基础研究项目(2017-ZJ-713);; 青海省高层次卫生人才(领军人才)项目(2018年,2019年);; 青海省重点研发与转化科技合作专项(2019-HZ-819)

作者(Author): 贾守宁;张庆光;马春花;李欣;王双玺;陈文娟;窦萱;葛广波;
JIA Shou-ning;ZHANG Qing-guang;MA Chun-hua;LI Xin;WANG Shuang-xi;CHEN Wen-juan;DOU Xuan;GE Guang-bo;Qinghai Hospital of Traditional Chinese Medicine;Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine;

Email:

DOI: 10.19540/j.cnki.cjcmm.20200323.402

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