中国中药杂志

2021, v.46(19) 5052-5063

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慢性心力衰竭气虚血瘀证及复方人参补气颗粒干预机制的蛋白质组学研究
Chronic heart failure due to Qi deficiency and blood stasis and intervention mechanism of Compound Renshen Buqi Granules:a proteomics-based study

吴昱杰;王智博;李瑛;王道平;苗兰;任钧国;潘映红;刘建勋;
WU Yu-jie;WANG Zhi-bo;LI Ying;WANG Dao-ping;MIAO Lan;REN Jun-guo;PAN Ying-hong;LIU Jian-xun;Graduate School, Beijing University of Chinese Medicine;Institute of Basic Medical Science, Xiyuan Hospital, China Academy of Chinese Medical Sciences;Institute of Crop Science, Chinese Academy of Agricultural Sciences;Beijing Key Laboratory of Pharmacology of Traditional Chinese Medicine;

摘要(Abstract):

复方人参补气颗粒广泛用于治疗慢性心力衰竭(chronic heart failure, CHF)气虚血瘀证,但其作用机制不明确。该研究基于定量蛋白质组学技术研究CHF气虚血瘀证发病及复方人参补气颗粒的干预机制,进一步挖掘其生物学基础。将SD大鼠分为空白对照组、正常给药组、模型组、模型给药组和阳性药对照组,采用左前降支冠状动脉结扎复合慢性睡眠剥夺诱导CHF气虚血瘀证大鼠模型,正常给药组和模型给药组给予复方人参补气颗粒,阳性药对照组给予缬沙坦。6周后采集血清,采用非靶向定量蛋白质组学数据依赖性采集(data dependent acquisition, DDA)技术,比较各组间蛋白表达差异,通过靶向定量蛋白质组学非数据依赖性采集(data independent acquisition, DIA)技术对差异表达蛋白(differentially expressed proteins, DEPs)进行验证。证候指标显示,与空白对照组相比,模型组大鼠体质量、抓力值、脉搏幅度均降低,舌面RGB值升高,病理形态学显示心肌组织发生炎细胞浸润、细胞变性坏死、组织纤维化等,经复方人参补气颗粒干预后多项指标发生回调。蛋白质组学结果显示,与空白对照组相比,模型组和正常给药组分别发现144、111个DEPs;与模型组相比,模型给药组和阳性药对照组分别发现107、194个DEPs;与正常给药组相比,模型给药组发现119个DEPs。经过DIA验证,6个蛋白在各组中的定量与DDA保持一致。证候指标和心肌组织病理形态学显示建立CHF气虚血瘀证模型,模型组的蛋白表达谱发生了明显的改变,并发现复方人参补气颗粒可逆转免疫蛋白等的差异表达来调控CHF气虚血瘀证,为CHF气虚血瘀证的发病机制和复方人参补气颗粒的干预机制研究提供线索。
Compound Renshen Buqi Granules have been widely used to treat chronic heart failure(CHF) due to Qi deficiency and blood stasis, but the mechanism of action remains unclear. This paper explored the pathogenesis of CHF due to Qi deficiency and blood stasis and the intervention mechanism of Compound Renshen Buqi Granules based on quantitative proteomics for uncovering the biological basis. SD rats were divided into the normal control(N) group, normal+Compound Renshen Buqi Granules(ND) group, model(M) group, model+Compound Renshen Buqi Granules(D) group, and positive control(Y) group. The rat model of CHF due to Qi deficiency and blood stasis was established by ligation of the left anterior descending(LAD) coronary artery and chronic sleep deprivation. The rats in the ND group and D group were provided with Compound Renshen Buqi Granules, while those in the Y group received valsartan. Six weeks later, the serum was sampled and the data-dependent acquisition(DDA) was employed for the non-targeted quantitative proteomics analysis of the differences in protein expression among groups, followed by the targeted analysis of differentially expressed proteins(DEPs) generated by data-independent acquisition(DIA). Compared with the N group, the rats in the M group pre-sented with decreased body weight, grip strength, and pulse amplitude and increased RGB value on the tongue surface. The pathomorphological examination revealed inflammatory cell infiltration, cell degeneration and necrosis, tissue fibrosis, etc. After the intervention with Compound Renshen Buqi Granules, multiple indicators were reversed. As demonstrated by proteomics results, there were 144 and 111 DEPs found in the M group and ND group in comparison with the N group. Compared with the M group, 107 and 194 DEPs were found in the D group and the Y group, respectively. Compared with the ND group, 119 DEPs were detected in the D group. As illustrated by DIA-based verification, the quantitative results of six proteins in each group were consistent with those by DDA. The syndrome indicators and pathomorphological examination results demonstrated that the protein expression profile of rats with CHF due to Qi deficiency and blood stasis changed obviously. However, Compound Renshen Buqi Granules were able to reverse the differential expression of immune proteins to regulate CHF of Qi deficiency and blood stasis syndrome, which has provided clues for figuring out the pathogenesis of CHF due to Qi deficiency and blood stasis and the intervention mechanism of Compound Renshen Buqi Granules.

关键词(KeyWords): 慢性心力衰竭;气虚血瘀证;病证结合;中药;蛋白质组学
chronic heart failure(CHF);Qi deficiency and blood stasis syndrome;disease-syndrome combination;traditional Chinese medicine;proteomics

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金重点项目(82030124);; 国家自然科学基金面上项目(81873041)

作者(Author): 吴昱杰;王智博;李瑛;王道平;苗兰;任钧国;潘映红;刘建勋;
WU Yu-jie;WANG Zhi-bo;LI Ying;WANG Dao-ping;MIAO Lan;REN Jun-guo;PAN Ying-hong;LIU Jian-xun;Graduate School, Beijing University of Chinese Medicine;Institute of Basic Medical Science, Xiyuan Hospital, China Academy of Chinese Medical Sciences;Institute of Crop Science, Chinese Academy of Agricultural Sciences;Beijing Key Laboratory of Pharmacology of Traditional Chinese Medicine;

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