脑心通胶囊药效成分对HUVEC增殖及其JAK/STAT信号通路、血管活性物质、黏附分子、炎症因子的影响
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篇名: 脑心通胶囊药效成分对HUVEC增殖及其JAK/STAT信号通路、血管活性物质、黏附分子、炎症因子的影响
TITLE: Effects of Active Components of Naoxintong Capsule on the Proliferation of HUVEC and Its JAK/STAT Signal Pathway,Vasoactive Substances ,Adhesion Molecules and Inflammatory Factors
摘要: 目的:研究脑心通胶囊主要药效成分对人脐静脉内皮细胞(HUVEC)增殖及其JAK/STAT信号通路、血管活性物质、黏附分子、炎症因子的影响,以期阐明脑心通胶囊活血化瘀的作用机制。方法:采用CCK-8法检测不同浓度的12个脑心通胶囊药效成分[咖啡酸(1.56~200μmol/L)、阿魏酸(1.56~200μg/mL)、洋川芎内酯H(3.125~200μmol/L)、丁烯基苯酞(3.125~200μmol/L)、藁本内酯(1.56~200μmol/L)、隐丹参酮(0.625~80μmol/L)、丹参素钠(1.56~200μmol/L)、芍药苷(1.56~200μmol/L)、芒柄花素(1.56~200μmol/L)、丹酚酸B(1.56~200μmol/L)、儿茶素(1.56~200μmol/L)、黄芪甲苷(1.56~200μmol/L)]作用24h后对HUVEC增殖的影响;采用逆转录-聚合酶链式反应(RT-PCR)法检测上述各药效成分(均分别设置3个剂量组,并设0μmol/L的空白对照组,下同)对HUVEC的JAK/STAT信号通路关键蛋白Janus激酶(JAK2)、信号转导子及转录激活子(STAT3)、蛋白激酶B(Akt)、细胞外调节蛋白激酶(ERK)mRNA表达的影响;采用酶联免疫吸附试验(ELISA)检测各药效成分对HUVEC的纤溶酶原激活物抑制因子1(PAI-1)、血管内皮细胞黏附分子1(VCAM-1)、细胞间黏附分子1(ICAM-1)、血管内皮生长因子(VEGF)、核因子κBp65(NF-κBp65)表达的影响。结果:阿魏酸(6.25、25~200μg/mL)、洋川芎内酯H(6.25~200μmol/L)、藁本内酯(200μmol/L)、隐丹参酮(10~80μmol/L)、芍药苷(1.56、6.25、12.5μmol/L)、丹酚酸B(1.56~12.5、200μmol/L)和儿茶素(25μmol/L)可显著抑制HU-VEC增殖,咖啡酸(1.56、12.5μmol/L)、藁本内酯(50μmol/L)、丹参素钠(6.25μmol/L)、芒柄花素(1.56、100、200μmol/L)可显著促进HUVEC细胞增殖(P<0.05或P<0.01)。与空白对照组比较,阿魏酸、芒柄花素、丹酚酸B和黄芪甲苷部分剂量组细胞中JAK2、STAT3和Akt的mRNA相对表达量均显著降低(P<0.05或P<0.01);咖啡酸、阿魏酸和丁烯基苯酞部分剂量组细胞中PAI-1表达水平显著降低,咖啡酸、阿魏酸、丁烯基苯酞、隐丹参酮、芒柄花素和儿茶素部分剂量组细胞中ICAM-1和VCAM-1表达水平显著降低,阿魏酸、丁烯基苯酞、芒柄花素、丹酚酸B和黄芪甲苷部分剂量组细胞中NF-κBp65表达水平显著降低,咖啡酸和儿茶素部分剂量组细胞中VEGF表达水平显著升高(P<0.05或P<0.01)。结论:脑心通胶囊中药效成分可能通过抑制HUVEC中JAK/STAT信号通路关键蛋白mRNA和PAI-1、ICAM-1、VCAM-1、NF-κBp65的表达,并促进VEGF表达,从而发挥其活血化瘀的功效。
ABSTRACT: OBJECTIVE:To study the effects of the main active components of Naoxintong capsule (NXTC)on the proliferation of human umbilical vein endothelial cell(HUVEC) and its key protein JAK/STAT signal pathway , vasoactive substances ,adhesion molecules and inflammatory factors so as to clarify the m echanism of NXTC for promoting blood circulation and removing blood stasis. METHODS :The effects of different concentration of 12 active components [caffeic acid(1.56-200 μmol/L),ferulic acid (1.56-200 μmol/L),senkyunolide H (3.125-200 μmol/L),n-butylidenephthalide(3.125-200 μmol/L),ligustilide(1.56-200 μmol/L),cryptotanshinone(0.625-80 μmol/L),tanshinol sodium (1.56-200 μmol/L),paeoniflorin (1.56-200 μmol/L),formononetin(1.56-200 μmol/L),salvianolic acid B (1.56-200 μmol/L),catechin(1.56-200 μmol/L)and astragaloside Ⅳ(1.56-200 μmol/L)] on the proliferation of HUVECs were evaluated by CCK- 8 assay. The effects of above active components(3 dose groups ,setting up 0 μmol/L blank control group,hereinafter)on mRNA expression of key proteins JAK 2, STAT3,Akt,ERK in JAK/STAT signal pathway were measured by RT-PCR. The effects of each active component on the expression of PAI- 1,VCAM-1,ICAM-1,VEGF and NF-κB p65 were detected by ELISA. RESULTS :Ferulic acid (6.25,25-200 μg/mL),senkyunolide H (6.25-200 μmol/L),ligustilide(200 μmol/L),cryptotanshinone(10-80 μmol/L),paeoniflorin(1.56, 6.25,12.5 μmol/L),salvianolic acid B (1.56-12.5 μmol/L,200 μmol/L)and catechin (25 μmol/L)could significantly inhibit the proliferation of HUVECs ;caffeic acid (1.56,12.5 μmol/L),ligustilide(50 μmol/L),trashinol sodium (6.25 μmol/L)and paeoniflorin(1.56,100,200 μmol/L)could significantly promote the proliferation of HUVECs (P<0.05 or P<0.01). Compared with blank control group ,mRNA expression of JAK 2,STAT3 and Akt were decreased significantly in some dose groups of ferulic acid,formononetin,salvianolic acid B and astragaloside Ⅳ(P<0.05 or P<0.01);the expression of PAI- 1 were significantly decreased in some dose groups of caffeic acid ,ferulic acid and n-butylphthalide;the expression of ICAM- 1 and VCAM- 1 were decreased significantly in some dose groups of caffeic acid ,ferulic acid ,n-butenylphthalide,cryptotanshinone,formononetin and catechin;the expression of NF-κB p65 were decreased significantly in some dose groups of ferulic acid ,n-butenylphthalide, formononetin,salvianolic acid B and astragaloside Ⅳ;the expression of VEGF were increased significantly in some dose groups of caffeic acid and catechin (P<0.05 or P<0.01). CONCLUSIONS :The active components of Naoxintong capsule may play the role of promoting blood circulation and removing blood stasis by inhibiting the expression of JAK/STAT signal pathway key protein mRNA and PAI- 1,ICAM-1,VCAM-1,NF-κB p65 in HUVEC ,and promoting the expression of VEGF.
期刊: 2021年第32卷第03期
作者: 李伟霞,王晓艳,贾文汇,张明亮,唐进法,李学林
AUTHORS: LI Weixia ,WANG Xiaoyan ,JIA Wenhui ,ZHANG Mingliang ,TANG Jinfa,LI Xuelin
关键字: 脑心通胶囊;人脐静脉内皮细胞;血管活性物质;黏附分子;炎症因子;药效成分;JAK/STAT信号通路;作用机制
KEYWORDS: Naoxintong capsule ; Human umbilical vein endothelial cells ;Vasoactive substances ;Adhesion molecule ;
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