摘要
目的探讨芍药苷抑制核因子-κB(NF-κB)/YY1信号通路改善肿瘤坏死因子-α(TNF-α)诱导的人脐静脉内皮细胞(HUVECs)炎症反应的机制。方法体外培养HUVECs,采用TNF-α(20 ng/mL)诱导的HUVECs炎症损伤模型,给予不同剂量的芍药苷(80μmol/L和160μmol/L)干预,采用细胞毒性活性检测试剂盒(CCK-8)检测各组细胞的活力;酶联免疫吸附实验(ELISA)法检测各组细胞上清液中白细胞介素-6(IL-6)和白细胞介素-8(IL-8)的含量;实时荧光定量聚合酶链反应(RT-PCR)检测各组细胞NF-κB和YY1基因表达;蛋白免疫印迹(Western Blot)法检测各组细胞NF-κB、YY1和环氧化酶-2(COX-2)蛋白表达。结果TNF-α可诱导HUVECs活力下降,提高炎性因子IL-6、IL-8的表达,促进炎症介质COX-2的活化,激活NF-κB/YY1信号通路;不同剂量的芍药苷能改善HUVECs的活力,降低炎性因子IL-6、IL-8和炎症介质COX-2的表达,抑制炎症信号通路NF-κB/YY1的活化。结论芍药苷通过抑制NF-κB/YY1信号通路改善内皮细胞的炎症反应,发挥抗动脉粥样硬化的作用。
Objective To explore the possible molecular mechanism of paeoniflorin(PF)protecting against TNF-α-induced inflammatory response in human umbilical vein cells(HUVECs)via NF-κB/YY1 signaling pathway.Methods HUVECs were cultured in vitro and pre-incubated 2 h with different concentration of PF(80μmol/L,160μmol/L)and followed by co-incubation with TNF-α(20 ng/mL)for 24 h.CCK-8 assay was used to examine cell viability.Enzyme-linked immunosorbent assays(ELISA)was performed to detect the concentration of IL-6 and IL-8.Real time polymerase chain reaction(RT-PCR)was conducted to measure the mRNA expression of NF-κB and YY1,and western blotting(WB)was conducted to measure protein level of NF-κB,YY1 and COX-2.Results The Results showed that TNF-α(20 ng/mL)impaired cell viabilities,induced overexpression of IL-6/IL-8,promoted the activation of NF-κB,YY1 and COX-2.Compared with modal group,different concentration of PF could increase the cell viability,decrease the expression of IL-6/IL-8 and COX-2,curb the activation of NF-κB,YY1.Conclusion PF suppressed TNF-α-mediated inflammatory response in HUVECs via NF-κB/YY1 signaling pathway and play an anti-atherosclerosis role.
作者
陈莉
范晓迪
史大卓
白瑞娜
CHEN Li;FAN Xiaodi;SHI Dazhuo;BAI Ruina(Institute of Cardiovascular Disease,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Clinical College of Traditional Chinese Medicine,Peking University(Xiyuan),Beijing 100191,China)
出处
《中西医结合心脑血管病杂志》
2020年第22期3756-3763,共8页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
基金
国家自然科学基金项目(No.81603489)
中央级公益性科研院所基本科研业务费专项基金(No.ZZ13-YQ-005)
中国中医科学院西苑医院“青师计划”。