天然产物研究与开发 ›› 2024, Vol. 36 ›› Issue (10): 1742-1750.doi: 10.16333/j.1001-6880.2024.10.011

• 开发研究 • 上一篇    下一篇

怀菊花茎叶中聚炔类化合物对ox-LDL诱导的HUVECs细胞损伤的保护作用

王小兰1,3,石静亚2,李   孟2,王滢清2,郑晓珂2,3,冯卫生2,3*   

  1. 1河南中医药大学中医药科学院;2河南中医药大学药学院;3河南省中药开发工程技术研究中心,郑州 450046
  • 出版日期:2024-10-28 发布日期:2024-10-28
  • 基金资助:
    河南省重大科技专项(171100310500);河南省博士后基金;河南省高等学校重点科研项目(21B360003);河南省中医药科学研究专项(20-21ZY2148)

Protective effect of polyacetylene compounds from the stem and leaves of Chrysanthemum morifolium Ramat on the HUVECs damage induced by ox-LDL

WANG Xiao-lan1,3,SHIJing-ya2,LI Meng2,WANG Ying-qing2,ZHENG Xiao-ke2,3,FENG Wei-sheng2,3*   

  1. 1Academy of Chinese Medical Sciences,Henan University of Chinese Medicine;2School of Pharmacy,Henan University of Chinese Medicine;3The Engineering and Technology Center for Chinese Medicine Development of Henan Province,Zhengzhou 450046,China
  • Online:2024-10-28 Published:2024-10-28

摘要:

研究怀菊花茎叶中分离得到的聚炔类化合物对氧化低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)损伤的保护作用及初步探讨作用机制。采用ox-LDL(200 μg/mL)处理HUVECs细胞建立内皮细胞损伤模型,同时给予聚炔类化合物1~10(100 μg/mL)干预;通过CCK-8法检测细胞生存率,筛选出有细胞保护作用的目标化合物;采用目标化合物59(12.5、25、50、100 μg/mL)干预细胞24 h,CCK-8检测细胞生存率,采用流式细胞术检测细胞凋亡率和活性氧(reactive oxygen species,ROS)生成率,酶联免疫(ELISA)检测细胞上清液中白介素-1β(interleukin-1β,IL-1β)、白介素-6(interleukin-6,IL-6)及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量;细胞免疫印迹法(In-Cell Western)检测细胞中核转录因子-κB(nuclear transcription factor-κB,NF-κB)2p100/p52 与NF-κBp65蛋白的相对表达水平。结果显示:与正常对照组(Con)相比,模型组(ox-LDL)中的细胞生存率明显下降,细胞凋亡率、ROS生成,炎性因子IL-1β、IL-6及TNF-α明显升高(P<0.01);与ox-LDL组相比,聚炔类化合物5和9可以明显提高HUVECs细胞的生存率、降低细胞凋亡率、ROS生成率,炎性因子IL-1β、IL-6及TNF-α的分泌(P<0.05 或P<0.01),下调NF-κB2 p100/p52、NF-κB p65蛋白的表达(P<0.05或P<0.01)。综上所述,怀菊花茎叶中聚炔苷类成分可以改善ox-LDL诱导的HUVECs细胞损伤与功能障碍,其作用机制可能与NF-κB信号通路相关。

关键词: 怀菊花茎叶, 聚炔类化合物, HUVECs, 凋亡与炎症, NF-κB

Abstract:

The aim of this research was to study the protective effect of polyacetylene compounds isolated from the stems and leaves of Chrysanthemum morifolium Ramat on human umbilical vein endothelial cells (HUVECs) damaged by oxidized low density lipoprotein (ox-LDL),and explore the preliminary mechanism of effect.HUVECs cells were treated with ox-LDL (200 μg/mL) to establish the endothelial cell injury model,and polyacetylene compounds 1-10 (100 μg/mL) were given at the same time.The cell survival rate was measured by CCK-8,and the target compounds with cell protection were screened.The compounds 5 and 9 (12.5,25,50,100 μg/mL) were used to treat the cells for 24 h,CCK-8 was used to detect the cell survival rate,and flow cytometry was used to detect the apoptosis rate and reactive oxygen species (ROS) production rate.The contents of inflammatory factors interleukin-1β (IL-1β),interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the cell culture fluid were detected by enzyme-linked immunosorbent assay (ELISA).In-Cell Western was performed to detect the relative expression levels of Nuclear transcription factor-κB (NF-κB) p100/p52 and NF-κBp65.The results showed:compared with the normal control group (Con),the cell survival rate in model group (ox-LDL) was significantly decreased,and the cell apoptosis rate,ROS production rate,inflammatory factors IL-1β,IL-6 and TNF-α were significantly increased (P<0.01).Compared with ox-LDL group,compounds 5 and 9 significantly increased the survival rate of HUVECs cells,reduced apoptosis rate,ROS production rate,and inflammatory factors IL-1β,IL-6 and TNF-α (P<0.05 or P<0.01),down-regulated the expression of NF-κB2 p100/p52 and NF-κB p65 proteins(P<0.05 or P<0.01).In conclusion,polyacetylene glycosides in stems and leaves of C. morifolium can improve ox-LDL induced HUVECs cell damage and dysfunction,and the mechanism of action may be related to NF-κB signaling pathway.

Key words:

中图分类号:  R285.5