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Application and Prospect of Platelet Multi-omics Technology in Study of Blood Stasis Syndrome

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Chinese Journal of Integrative Medicine 2022年第2期28卷 99-105页

作者： LI Ying SUN Ming-qian LI Lei ZHANG Ye-hao MIAO Lan liu jian-xun Beijing Key Laboratory of Pharmacology of Chinese Materia Region Xiyuan HospitalChina Academy of Chinese Medical SciencesNational Clinical Research Center for Chinese Medicine CardiologyBeijing(100091)China

The abnormality of platelet function plays an important role in the pathogenesis and evolution of blood stasis syndrome(BSS).The explanation of its mechanism is a key scientific issue in the study of cardiovascular and cerebrovascular diseases and treatment.System biology technology provides a good technical platform for further development of platelet multi-omics,which is conducive to the scientific interpretation of the biological mechanism of BSS.The article summarized the pathogenesis of platelets in BSS,the mechanism of action of blood activating and stasis resolving drugs,and the application of genomics,proteomics,and metabonomics in platelet research,and put forward the concept of"plateletomics in BSS".Through the combination and cross-validation of multi-omics technology,it mainly focuses on the clinical and basic research of cardiovascular and cerebrovascular diseases;through the interactive verification of multi-omics technology and system biology,it mainly focuses on the platelet function and secretion system.The article systematically explains the molecular biological mechanism of platelet activation,aggregation,release,and other stages in the formation and development of BSS,and provides a new research idea and method for clarifying the pathogenesis of BSS and the mechanism of action of blood activating and stasis resolving drugs.

关键词： blood stasis syndrome platelet transcriptomics metabonomics proteomics

Huoxin Pill Reduces Myocardial Ischemia Reperfusion Injury in Rats via TLR4/NFκB/NLRP3 Signaling Pathway

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Chinese Journal of Integrative Medicine 2023年第12期29卷 1066-1076页

作者： CAO Ce QI Yu-tong WANG Ao-ao WANG Zi-yan liu Zi-xin MENG Hong-xu LI Lei liu jian-xun Institute of Basic Medical Sciences of Xiyuan Hospital China Academy of Chinese Medical SciencesBeijing Key Laboratory of Chinese Materia PharmacologyNational Clinical Research Center of Traditional Chinese Medicine for Cardiovascular DiseasesBeijing(100091)China Institute of Chinese Medicine Sciences Guangdong Pharmaceutical UniversityGuangzhou(510006)China

Objective:To explore the protective effect of Huoxin Pill(HXP)on acute myocardial ischemia-reperfusion(MIRI)injury in rats.Methods:Seventy-five adult SD rats were divided into the sham-operated group,model group,positive drug group(diltiazem hydrochloride,DH),high dose group(24 mg/kg,HXP-H)and low dose group(12 mg/kg,HXP-L)of Huoxin Pill(n=15 for every group)according to the complete randomization method.After 1 week of intragastric administration,the left anterior descending coronary artery of the rat's heart was ligated for 45 min and reperfused for 3 h.Serum was separated and the levels of creatine kinase(CK),creatine kinase isoenzyme(CK-MB)and lactate dehydrogenase(LDH),superoxide dismutase(SOD),and malondialdehyde(MDA),hypersensitive C-reactive protein(hs-CRP)and interleukin-1β(IL-1β)were measured.Myocardial ischemia rate,myocardial infarction rate and myocardial no-reflow rate were determined by staining with Evans blue and 2,3,5-triphenyltetrazolium chloride(TTC).Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN)databases were used to screen for possible active compounds of HXP and their potential therapeutic targets;the results of anti-inflammatory genes associated with MIRI were obtained from GeneC ards,Drugbank,Online Mendelian Inheritance in Man(OMIM),and Therapeutic Target Datebase(TTD)databases was performed;Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment were used to analyze the intersected targets;molecular docking was performed using AutoD ock Tools.Western blot was used to detect the protein expression of Toll-like receptor 4(TLR4)/nuclear factor kappa-B(NFκB)/NOD-like receptor protein 3(NLRP3).Results:Compared with the model group,all doses of HXP significantly reduced the levels of LDH,CK and CK-MB(P<0.05,P<0.01);HXP significantly increased serum activity of SOD(P<0.05,P<0.01);all doses of HXP significantly reduced the levels of hs-CRP

关键词： Houxin Pill myocardial ischemia-reperfusion injury TLR4/NFκB/NLRP3 signaling pathway network pharmacology molecular docking

巨噬细胞极化在肺部疾病中的作用机制及中药的干预

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中国中药杂志 2024年第2期49卷 334-343页

作者：梁晓刘建勋中国中医科学院西苑医院基础医学研究所国家中医心血管病研究中心中药药理北京市重点实验室北京100091

巨噬细胞作为机体重要的免疫细胞,是参与维持肺内微环境稳态的关键因素,由于其高度的可塑性,可在不同的条件下极化为不同的亚型:经典活化巨噬细胞M1和选择性活化巨噬细胞M2,两者分别具有促炎和抗炎作用。M1/M2表型与肺部疾病的发生发展... 详细信息

巨噬细胞作为机体重要的免疫细胞,是参与维持肺内微环境稳态的关键因素,由于其高度的可塑性,可在不同的条件下极化为不同的亚型:经典活化巨噬细胞M1和选择性活化巨噬细胞M2,两者分别具有促炎和抗炎作用。M1/M2表型与肺部疾病的发生发展密切相关,在巨噬细胞极化过程中涉及的多种信息分子和细胞因子在各种肺部疾病中均有重要的调控表型的作用,且在不同疾病中表型的转换也存在明显差异。该文主要通过综述巨噬细胞极化生物学特性,探讨巨噬细胞极化在支气管哮喘、慢性阻塞性肺疾病、急性肺损伤、肺纤维化等肺部疾病中的作用特点和以调控巨噬细胞极化为靶点的中药单体及中药复方对相关疾病的研究进展,旨在挖掘巨噬细胞极化在调控肺部炎症方面的潜力,为相关临床研究提供新思路。

关键词：巨噬细胞极化 M1型巨噬细胞 M2型巨噬细胞肺部疾病中药

Salvianolic Acid A Protects Neonatal Cardiomyocytes against Hypoxia/Reoxygenation-Induced Injury by Preserving Mitochondrial Function and Activating Akt/GSK-3β Signals

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Chinese Journal of Integrative Medicine 2019年第1期25卷 23-30页

作者： LI Xue-li FAN Ji-ping liu jian-xun LIANG Li-na Institute of Basic Medical Sciences Xiyuan Hospital China Academy of Chinese Medical Sciences Experimental Research Center China Academy of Chinese Medical Sciences Post-doctoral Research Station China Academy of Chinese Medical Sciences China Academy of Chinese Medical Sciences Eye Function Laboratory Yanke Hospital China Academy of Chinese Medical Sciences

Objective: To investigate the effects of salvianolic acid A(SAA) on cardiomyocyte apoptosis and mitochondrial dysfunction in response to hypoxia/reoxygenation(H/R) injury and to determine whether the Akt signaling pathway might play a role. Methods: An in vitro model of H/R injury was used to study outcomes on primary cultured neonatal rat cardiomyocytes. The cardiomyocytes were treated with 12.5, 25, 50 μg/m L SAA at the beginning of hypoxia and reoxygenation, respectively. Adenosine triphospate(ATP) and reactive oxygen species(ROS) levels were assayed. Cell apoptosis was evaluated by flow cytometry and the expression of cleavedcaspase 3, Bax and Bcl-2 were detected by Western blotting. The effects of SAA on mitochondrial dysfunction were examined by determining the mitochondrial membrane potential(△Ψm) and mitochondrial permeability transition pore(m PTP), followed by the phosphorylation of Akt(p-Akt) and GSK-3β(p-GSK-3β), which were measured by Western blotting. Results: SAA significantly preserved ATP levels and reduced ROS production. Importantly, SAA markedly reduced the number of apoptotic cel s and decreased cleaved-caspase 3 expression levels, while also reducing the ratio of Bax/Bcl-2. Furthermore, SAA prevented the loss of △Ψm and inhibited the activation of m PTP. Western blotting experiments further revealed that SAA significantly increased the expression of p-Akt and p-GSK-3β, and the increase in p-GSK-3β expression was attenuated after inhibition of the Akt signaling pathway with LY294002. Conclusion: SAA has a protective effect on cardiomyocyte H/R injury; the underlying mechanism may be related to the preservation of mitochondrial function and the activation of the Akt/GSK-3β signaling pathway.

关键词： salvianolic acid A cardiomyocyte hypoxia/reoxygenation injury mitochondria Akt/GSK-3β

天然药物调控星形胶质细胞促进缺血性脑卒中后神经修复的研究进展

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中国药理学通报 2024年第6期40卷 1013-1019页

作者：黄子珊张业昊刘建勋广东药科大学中医药研究员院广东广州510006 中国中医科学院西苑医院基础医学研究所中药药理北京市重点实验室北京100091

星形胶质细胞在治疗缺血性脑卒中过程中起到了不可忽视的作用。我国拥有丰厚资源的天然药物,近年来针对星形胶质细胞的天然药物研究逐年增多,为治疗缺血性脑卒中提供了丰富的依据及思路。因此,该文对星形胶质细胞在缺血性脑卒中后对神... 详细信息

星形胶质细胞在治疗缺血性脑卒中过程中起到了不可忽视的作用。我国拥有丰厚资源的天然药物,近年来针对星形胶质细胞的天然药物研究逐年增多,为治疗缺血性脑卒中提供了丰富的依据及思路。因此,该文对星形胶质细胞在缺血性脑卒中后对神经修复的影响及机制,以及天然药物对该过程的调控从而促进修复的作用作一阐述。

关键词：星形胶质细胞神经修复天然药物缺血性脑卒中中药复方综述

GABAergic网络在神经发育和脑疾病中对髓鞘再生的研究进展

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中国药理学通报 2024年第5期40卷 801-806页

作者：范晓迪张业昊刘建勋中国中医科学院西苑医院基础医学研究所中药药理北京市重点实验室北京100091

少突胶质细胞前体细胞(oligodendrocyte precursor cells,OPCs)分化为成熟的少突胶质细胞(oligodendrocytes,OLs)是中枢神经系统轴突髓鞘形成和脱髓鞘疾病髓鞘再生的关键事件。研究表明,神经递质γ-氨基丁酸(γ-aminobutyric acid,GABA)... 详细信息

少突胶质细胞前体细胞(oligodendrocyte precursor cells,OPCs)分化为成熟的少突胶质细胞(oligodendrocytes,OLs)是中枢神经系统轴突髓鞘形成和脱髓鞘疾病髓鞘再生的关键事件。研究表明,神经递质γ-氨基丁酸(γ-aminobutyric acid,GABA)、GABAergic突触及其网络调控对少突神经胶质细胞的增殖、分化、迁移及髓鞘形成具有重要的调节作用。因此,该文总结了近些年关于GABA及其受体在少突胶质细胞谱系中发挥的生物学功能,GABAergic中间神经元与OPCs之间的交互作用及其介导髓鞘形成的网络调控和未来潜在的候选药物的研究。基于此了解控制OLs分化机制对于确定促进髓鞘修复的治疗策略至关重要,围绕GABAergic的研究可能对开发脱髓鞘疾病的新型修复疗法具有潜在意义。

关键词： γ-氨基丁酸 GABA能网络少突胶质细胞中间神经元髓鞘再生神经发育与疾病

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延胡索乙素通过ULK1/FUNDC1通路抑制线粒体自噬减轻H9c2心肌细胞缺氧/复氧损伤

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中国中药杂志 2024年第5期49卷 1286-1294页

作者：辛高杰刘子馨陈原原张会雨郭帆彭涵李磊韩笑刘建勋付建华中国中医科学院西苑医院基础医学研究所北京100091 国家中医心血管病临床医学研究中心北京100091

该研究探讨了延胡索乙素(tetrahydropalmatine,THP)通过UNC-51样激酶1(UNC-51-like kinase 1,ULK1)/FUN14结构域蛋白1(FUN14 domain containing 1,FUNDC1)通路抑制线粒体自噬减轻H9c2心肌细胞缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的具体机制。该研究以H9c2心肌细胞为研究对象,构建心肌细胞H/R损伤模型。首先采用细胞活力检测试剂盒检测细胞活性、微量法检测乳酸脱氢酶(lactate dehydrogenase,LDH)漏出量评估THP对H9c2心肌细胞H/R损伤的保护作用。然后采用化学荧光法检测细胞内活性氧、线粒体内活性氧、线粒体膜电位、自噬小体情况,并用萤光素法检测细胞内三磷酸腺苷(adenosine 5′-triphosphate,ATP)含量评估THP对线粒体的保护作用。进一步用免疫印记检测微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)膜型(LC3-Ⅱ)和包浆型(LC3-Ⅰ)的比值,活化的半胱氨酸天冬氨酸蛋白水解酶-3(cleaved caspase-3)表达水平、ULK1表达水平及其在Ser555位点的磷酸化程度,FUNDC1表达水平及其在Ser17位点的磷酸化程度探索其具体机制。结果显示,THP有效减轻H9c2细胞H/R后线粒体损伤,THP通过降低细胞内和线粒体内活性氧水平、升高线粒体膜电位保护线粒体,进而增加细胞ATP生成、升高细胞活性、降低细胞LDH漏出,减轻H9c2细胞H/R损伤;进一步研究发现THP能够降低自噬小体生成,降低LC3-Ⅱ/LC3-Ⅰ比值,降低凋亡相关蛋白cleaved caspase-3表达,表明THP通过抑制自噬减轻细胞凋亡;深入研究发现THP通过降低ULK1在Ser555位点和FUNDC1在Ser17位点的磷酸化程度,抑制线粒体自噬ULK1/FUNDC1通路激活,抑制线粒体自噬发生。ULK1激动剂BL-918的应用逆向验证了THP降低ULK1和FUNDC1磷酸化的作用。综上所述,THP通过ULK1/FUNDC1通路抑制线粒体自噬减轻H9c2心肌细胞H/R损伤。

关键词：延胡索乙素线粒体自噬 H9c2心肌细胞缺氧/复氧损伤 ULK1/FUNDC1通路

Mechanism of'Invigorating Qi and Promoting Blood Circulation'Drug Pair Ginseng-Danshen on Treatment of Ischemic Heart Disease Based on Network Pharmacology

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Chinese Journal of Integrative Medicine 2021年第6期27卷 440-445页

作者： XIN Gao-jie ZHAO Yu-wei LI Ling-mei JIA Fei-fan HAN Xiao LI Lei GUO Hao MENG Hong-xu FU jian-hua liu jian-xun Institute of Basic Medical Sciences of Xiyuan Hospital China Academy of Chinese Medical SciencesBeijing Key Laboratory of Pharmacology of Chinese MateriaBeijing(100091)China National Clinical Research Center for Chinese Medicine Cardiology Xiyuan HospitalChina Academy of Chinese Medical SciencesBeijing(100091)China

Objective:Using network pharmacology to explore the mechanism of the'invigorating qi and promoting blood circulation'drug pair Ginseng-Danshen(Salvia miltiorrhiza)on treatment of ischemic heart disease(IHD).Methods:The chemical constituents of ginseng and Danshen drug pair were identified by searching the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),and the potential targets of the pair were identified.The pharmacodynamics of the pair was analyzed using network pharmacology.The targets of IHD were identified by database screening.Using protein-protein interaction network,the interaction targets of Ginseng-Danshen on IHD were constructed.A"constituent-target-disease"interaction network was constructed using Cytoscape software,Gene Ontology(GO)term enrichment analysis and biological pathway enrichment analysis were carried out,and the mechanism of improving myocardial ischemia by the Ginseng-Danshen drug pair was investigated.Results:Seventeen active constituents and 53 targets were identified from ginseng,53 active constituents and 61 targets were identified from Danshen,and 32 protein targets were shared by ginseng and Danshen.Twenty GO terms were analyzed,including cytokine receptor binding,cytokine activity,heme binding,and antioxidant activity.Sixty Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathways were analyzed,including phosphatidylinositol 3-kinase-serine-threonine kinase(PI3 K-AKT)signaling pathway,p53 signaling pathway,interleukin 17 signaling pathway,tumor necrosis factor signaling pathway,and the advanced glycation end product(AGE)-the receptor for AGE(RAGE)signaling pathway in diabetic complications.Conclusion:The specific mechanism of Ginseng-Danshen drug pair in treating IHD may be associated with improving the changes of metabolites inbody,inhibiting the production of peroxides,removing the endogenous oxygen free radicals,regulating the expression of inflammatory factors,reducing myocardial cell apoptosis and promoting vascular regeneration.

关键词： network pharmacology Ginseng-Danshen Salvia miltiorrhiza drug pair ischemic heart disease invigorating qi and promoting blood circulation

人参皂苷Re通过Nrf2/HO-1/PGC-1α通路调控线粒体生物发生减轻H9c2心肌细胞缺氧/复氧损伤的研究

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中国中药杂志 2024年第4期49卷 1064-1072页

作者：辛高杰陈原原刘子馨胥淑娟张会雨郭帆彭涵李磊韩笑刘建勋付建华中国中医科学院西苑医院基础医学研究所北京100091 国家中医心血管病临床医学研究中心北京100091

该文探讨了人参皂苷Re通过核因子E2相关因子2(nuclear factor-erythroid 2-related factor 2,Nrf2)/血红素氧合酶-1(heme oxidase-1,HO-1)/过氧化物增殖体激活受体γ共激活因子-1α(PGC-1α)调控线粒体生物发生减轻H9c2细胞缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的机制。将H9c2细胞缺氧培养4 h再复氧培养2 h构建心肌细胞H/R损伤模型。Re预给药干预后,首先检测细胞活性、超氧化物歧化酶(superoxide dismutase,SOD)活性、丙二醛(malondialdehyde,MDA)含量、细胞内活性氧(Cyto-ROS)和线粒体内活性氧(Mito-ROS)水平,评估Re通过抗氧化应激对H9c2细胞H/R损伤的保护作用。其次通过荧光探针检测线粒体膜电位(mitochondrial membrane potential,ΔΨ_(m))、线粒体膜通透性开放孔(membrane permeability open pore,mPTP)变化情况,通过免疫荧光检测TOM20表达水平,研究Re对线粒体的保护作用。进一步用免疫印迹检测caspase-3、cleaved caspase-3、Cyto C、Nrf2、HO-1、PGC-1α蛋白表达水平,探讨Re抗氧化应激保护线粒体减轻H/R损伤的具体机制。结果显示,与模型组相比,Re有效减轻H9c2细胞H/R损伤氧化应激反应,Re可显著升高细胞中SOD活性,降低MDA含量,降低Cyto-ROS和Mito-ROS水平;Re对线粒体有较好保护作用,升高ΔΨ_(m),降低mPTP开放,升高TOM20表达;进一步研究表明Re促进Nrf2、HO-1、PGC-1α蛋白表达,降低凋亡相关调节因子caspase-3向cleaved caspase-3活化、Cyto C蛋白表达。综上所述,人参皂苷Re减轻H9c2细胞H/R损伤的具体机制与通过Nrf2/HO-1/PGC-1α通路促进线粒体生物发生,进而升高线粒体数量、改善线粒体功能,增强细胞抗氧化应激能力,减轻细胞凋亡有关。

关键词：人参皂苷Re 线粒体生物发生 Nrf2/HO-1/PGC-1α 缺氧/复氧损伤氧化应激