基于蛋白组学探讨龙生蛭胶囊抗动脉粥样硬化作用机制  

Mechanism of Longshengzhi Capsules(龙生蛭胶囊)against Atherosclerosis Based on Proteomics

作　　者：方欢乐[1] 姜欢欢 周亚明 王礼丰 陈衍斌 FANG Huanle;JIANG Huanhuan;ZHOU Yaming;WANG Lifeng;CHEN Yanbin(Medical School,Xi’an Peihua University,Xi’an 710125;Scientific Research Department of Buchang Pharma Co.,Ltd,Xi’an 710075;Xi’an Jiaotong University Health Science Center,Xi’an 710061)

机构地区：[1]西安培华学院医学院,西安710125 [2]陕西步长制药有限公司科研部,西安710075 [3]西安交通大学医学部,西安710061

出　　处：《中药药理与临床》2023年第4期13-18,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：陕西省重点研发计划项目(编号:2021SF-362);国家级大学生创新创业项目(编号:S202111400002)。

摘　　要：目的:探讨龙生蛭胶囊对动脉粥样硬化(AS)ApoE^(-/-)小鼠的干预作用及可能机制。方法:ApoE^(-/-)雄性小鼠分为模型对照组、龙生蛭胶囊1.8 g/kg组,每组10只,另取10只C57BL/6小鼠作为正常对照组;高脂饮食喂养ApoE^(-/-)小鼠8 w建立动脉粥样硬化模型。生化法检测小鼠血脂及MPO含量;HE染色观察主动脉病理形态变化;Western Blot检测主动脉TGF-β、Smad3蛋白表达;4D-Label-free定量蛋白质组学技术分析各组小鼠主动脉血管差异蛋白的表达;GO注释和KEGG通路富集分析相关通路和蛋白;平行反应监测(PRM)验证典型特异蛋白表达。结果:造模8 w后,与正常对照组比较,模型对照组血脂TC、TG、LDL含量升高,HDL含量降低(P<0.05或P<0.01);HE染色可见主动脉壁内膜明显增厚;血管组织中MPO、TGF-β、Smad3蛋白表达显著上调;与模型对照组相比较,龙生蛭胶囊1.8 g/kg组血脂降低,内膜损伤减轻,MPO、TGF-β、Smad3蛋白表达明显下调(P<0.05或P<0.01)。蛋白定量分析显示,与正常对照组相比,模型对照组有213个差异蛋白,其中上调177个,下调36个;与模型对照组比较,龙生蛭胶囊1.8 g/kg组共筛选出74个差异蛋白,其中42个表达上调,32个表达下调。这些差异蛋白主要参与了JAK2-STAT3信号通路、TGF-β信号通路、半乳糖代谢、RAP1信号通路、鞘磷脂信号通路、等。龙生蛭胶囊1.8 g/kg组可显著下调CDK9、MPO、SAA2、MSR1、FBN1、等蛋白表达,显著上调ABCA1等蛋白表达。结论:龙生蛭胶囊具有一定的抗动脉粥样硬化作用,其可能主要通过调控动脉粥样硬化ApoE^(-/-)小鼠炎症、脂质代谢相关蛋白及通路改善动脉粥样硬化。Objective:To investigate the effect of Longshengzhi Capsules(龙生蛭胶囊,LSZCs)on ApoE^(-/-)mice with atherosclerosis(AS)and the mechanism.Methods:ApoE^(-/-)mice were divided into a model group and a 1.8 g/kg LSZC treatment group,with 10 mice in each group,and another 10 C57BL/6 mice were taken as a normal control group.ApoE^(-/-)mice were fed with high-fat diet for 8 weeks to establish AS model.Biochemical method was applied to detect the serum lipid and myeloperoxidase(MPO)content.The hematoxylin-eosin(HE)staining was used to observe the pathological changes in aorta.Western blotting was employed to determine the protein expression levels of transforming growth factor-β(TGF-β)and Smad3.4D-label-free quantitative proteomics technology was used to screen out the differential expressed proteins in the aortic vessels.Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed for the differential proteins.The typical specific protein expression was verified by parallel reaction monitoring(PRM).Results:After 8 weeks of modeling,as compared with normal control group,the content of total cholesterol(TC),triglyceride(TG),and low-density lipoprotein(LDL)was increased in the model group,while the content of high-density lipoprotein(HDL)was decreased(P<0.05 or P<0.01).HE staining showed a significant increase in the intima of the aortic wall.The protein expression levels of MPO,TGF-β,and Smad3 were significantly up-regulated in vascular tissues.As compared with the model group,the blood lipids and intimal injury were decreased in the 1.8 g/kg LSZC group,and the protein expression levels of MPO,TGF-β,and Smad3 were down-regulated significantly(P<0.05 or P<0.01).Quantitative protein analysis showed that as compared with the normal control group,213 differential proteins were screened out in the model group,in which 177 were up-regulated and 36 were down-regulated.As compared with the model group,74 differential proteins were screened out

关 键 词：龙生蛭胶囊 ApoE^(-/-)小鼠 动脉粥样硬化 多肽体外标记定量技术 作用机制 

分 类 号：R285.5[医药卫生—中药学]