芒柄花黄素调节TLR4/NF-κB通路对ox-LDL诱导的人冠状动脉内皮细胞损伤的影响  

Effect of Formononetin on Human Coronary Endothelial Cell Against ox-LDL-induced Injury by Regulating TLR4/NF-κB Pathway

作　　者：袁小翠 廖锦钰 付丽 罗勇[2] 陈东 YUAN Xiaocui;LIAO Jinyu;FU Li;LUO Yong;CHEN Dong(Mianyang Central Hospital,Mianyang 621000,Sichuan,China;The Third People′s Hospital of Chengdu;Mianyang People′s Hospital)

机构地区：[1]绵阳市中心医院,四川绵阳621000 [2]成都市第三人民医院 [3]绵阳市人民医院

出　　处：《中西医结合心脑血管病杂志》2025年第7期1015-1021,共7页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：四川省卫生健康委员会科研项目(No.19PJ011)。

摘　　要：目的:探讨芒柄花黄素(Form)调节Toll样受体4(TLR4)/核因子κB(NF-κB)通路对氧化型低密度脂蛋白(ox-LDL)诱导的人冠状动脉内皮细胞(HCAECs)损伤的影响。方法:采用24μg/mL的ox-LDL处理HCAECs 40 h诱导损伤和炎症,未用ox-LDL处理的HCAECs作为对照组。将ox-LDL处理的HCAECs分为HCAECs组、L-Form组(16μmol/L Form)、M-Form组(32μmol/L Form)、H-Form组(64μmol/L Form)、脂多糖(LPS)组(5μg/mL TLR4/NF-κB p65通路激活剂LPS)、H-Form+LPS组(64μmol/L Form+5μg/mL LPS)。采用细胞计数(CCK-8)法检测细胞增殖活性;流式细胞术检测细胞凋亡;Transwell法测定细胞迁移和侵袭;基质胶评估管形成能力;酶联免疫吸附试验(ELISA)检测白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平;蛋白免疫印迹法(Western Blot)检测TLR4/NF-κB通路蛋白表达水平。结果:与对照组相比,HCAECs组凋亡率,IL-1β、IL-6、TNF-α水平,TLR4、p-NF-κB p65/NF-κB p65蛋白表达明显升高(P<0.05),细胞增殖活性OD值明显降低(P<0.05),迁移细胞数量、侵袭细胞数量、毛细管管样的节点数量明显减少(P<0.05),管总长度明显缩小(P<0.05);与HCAECs组相比,L-Form组、M-Form组、H-Form组HCAECs细胞凋亡率,IL-1β、IL-6、TNF-α水平,TLR4、p-NF-κB p65/NF-κB p65蛋白表达明显下降(P<0.05),细胞增殖活性OD值、迁移细胞数量、侵袭细胞数量、毛细管管样的节点数量、管总长度明显增加(P<0.05),而LPS组趋势相反;LPS消除了高浓度Form对HCAECs细胞有利的影响。结论:Form可能通过抑制TLR4/NF-κB信号通路来缓解ox-LDL诱导的HCAECs损伤。ObjectiveObjective:To investigate the effect of formononetin(Form)in regulating toll-like receptor 4(TLR4)/nuclear factorκB(NF-κB)pathway on human coronary endothelial cell(HCAECs)against oxidative low-density lipoprotein(ox-LDL)-induced injury.MethodsMethods:The HCAECs treated with 24μg/mL ox-LDL for 40 h induced injury and inflammation,and the non-ox-LDL treated HCAECs served as the control group.HCAECs treated with ox-LDL were divided into HCAECs group,L-form group(16μmol/L Form),M-Form group(32μmol/L Form),H-Form group(64μmol/L Form)and lipopolysaccharide(LPS)group(5μg/mL TLR4/NF-κB p65 pathway activator LPS)and H-Form+LPS groups(64μmol/L Form+5μg/mL LPS).Cell proliferation activity was detected by cell counting(CCK-8).Flow cytometry was used to detect apoptosis.Cell migration and invasion were measured by Transwell method.Matrix glue was used to evaluate tube formation ability.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).Western Blot was used to detect TLR4/NF-κB pathway protein expression.ResultsResults:Compared with the control group,the apoptosis rate,the levels of IL-1β,IL-6,TNF-α,the expression of TLR4,P-NF-κB p65/NF-κB p65 protein in HCAECs group significantly increased(P<0.05),and the OD value of cell proliferation activity significantly decreased(P<0.05),the number of migrating cells,the number of invading cells,the number of capillary tube nodes significantly decreased(P<0.05),and the total tube length significantly reduced(P<0.05).Compared with HCAECs group,the apoptosis rate of HCAECs cells,the levels of IL-1β,IL-6,and TNF-α,and the expression of TLR4 and P-NF-κB p65/NF-κB p65 protein in L-Form,M-Form,and H-Form groups significantly decreased(P<0.05).OD value of cell proliferation activity,number of migrating cells,number of invading cells,number of capillary tube nodes,and total tube length significantly increased(P<0.05),but the trend was opposite in LPS group.LPS elimina

关 键 词：芒柄花黄素 人冠状动脉内皮细胞 氧化型低密度脂蛋白 Toll样受体4/核因子κB信号通路 实验研究 

分 类 号：R285[医药卫生—中药学] R285.5[医药卫生—中医学]