脑心通胶囊药效成分对人脐静脉内皮细胞JAK/STAT信号通路、血管活性物质、黏附分子、炎症因子的影响

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)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...

Effects of Active Components of Naoxintong Capsule on the JAK/STAT Signal Pathway,Vasoactive Substances,Adhesion Molecules and Inflammatory Factors of HUVEC

OBJECTIVE:To study the effects of the main active components of Naoxintong capsule(NXTC)on the JAK/STAT signal pathway of human umbilical vein endothelial cell(HUVEC)vasoactive substances,adhesion molecules and inflammatory factors so as to clarify the mechanism of NXTC for promoting blood circulation and removing blood stasis.METHODS:The effects of different concentration of 12 active componentscaffeic acid(1.56-200μmol/L),ferulic acid(1.56-200μmol/L),senkyunolide H(3.125-200μmol/L),n-butylidenephthalide(3.125-200μmol/L),ligustilide(1.56-200μmol/L),cryptotanshinone(0.625-80μmol/L),tanshinol sodium(1.56-200μmol/L),paeoniflorin(1.56-200μmol/L),formononetin(1.56-200μmol/L),salvianolic acid B(1.56-200μmol/L),catechin(1.56-200μmol/L)and astragalosideⅣ(1.56-200μmol/L)]on the proliferation of HUVECs were evaluated by CCK-8 assay.The effects of above active components(3 dose groups,setting up 0μmol/L blank control group,hereinafter)on mRNA expression of key proteins JAK2,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(6.25,25-200μg/mL),senkyunolide H(6.25-200μmol/L),ligustilide(200μmol/L),cryptotanshinone(10-80μmol/L),paeoniflorin(1.56,6.25,12.5μmol/L),salvianolic acid B(1.56-12.5μmol/L,200μmol/L)and catechin(25μmol/L)could significantly inhibit the proliferation of HUVECs;caffeic acid(1.56,12.5μmol/L),ligustilide(50μmol/L),trashinol sodium(6.25μmol/L)and paeoniflorin(1.56,100,200μmol/L)could significantly promote the proliferation of HUVECs(P<0.05 or P<0.01).Compared with blank control group,mRNA expression of JAK2,STAT3 and Akt were decreased significantly in some dose groups of ferulic acid,formononetin,salvianolic acid B and astragalosideⅣ(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 astragalosideⅣ;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.