Si Yanhong, Guo Shoudong, Fang Yongqi, Qin Shucun, Li Furong, Zhang Ying, Jiao Peng, Zhang Chunduo, Gao Linlin
College of Basic Medical Sciences, Taishan Medical University, Shandong, China.
Am J Chin Med. 2015;43(3):443-55. doi: 10.1142/S0192415X15500287. Epub 2015 Apr 27.
Oxidized low-density lipoprotein (ox-LDL)-induced macrophage foam cell formation and injury is one of the major atherogenic factors. This study is aimed to investigate the protective effect of celery seed extract (CSE) on ox-LDL-induced injury of macrophages and the underlying signaling pathway. RAW264.7 macrophages were pre-incubated with CSE for 24 h, followed by stimulation with ox-LDL. Oil red O staining and enzymatic colorimetry indicated CSE significantly lessened lipid droplets and total cholesterol (TC) content in ox-LDL-injured macrophages. ELISA revealed that CSE decreased the secretion of inflammatory cytokine TNF-α and IL-6 by 12-27% and 5-15% respectively. MTT assay showed CSE promoted cell viability by 16-40%. Cell apoptosis was also analyzed by flow cytometry and laser scanning confocal microscope and the data indicated CSE inhibited ox-LDL-induced apoptosis of macrophages. Meanwhile, western blot analysis showed CSE suppressed NF-κBp65 and notch1 protein expressions stimulated by ox-LDL in macrophages. These results suggest that CSE inhibits ox-LDL-induced macrophages injury via notch1/NF-κB pathway.
氧化型低密度脂蛋白（ox-LDL）诱导的巨噬细胞泡沫细胞形成和损伤是主要的致动脉粥样硬化因素之一。本研究旨在探讨芹菜籽提取物（CSE）对ox-LDL诱导的巨噬细胞损伤的保护作用及其潜在的信号通路。将RAW264.7巨噬细胞与CSE预孵育24小时，然后用ox-LDL刺激。油红O染色和酶比色法表明，CSE显著减少了ox-LDL损伤巨噬细胞中的脂滴和总胆固醇（TC）含量。酶联免疫吸附测定（ELISA）显示，CSE分别使炎性细胞因子肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的分泌降低了12%-27%和5%-15%。噻唑蓝（MTT）法检测显示，CSE使细胞活力提高了16%-40%。还通过流式细胞术和激光扫描共聚焦显微镜分析细胞凋亡，数据表明CSE抑制了ox-LDL诱导的巨噬细胞凋亡。同时，蛋白质免疫印迹分析显示，CSE抑制了ox-LDL刺激巨噬细胞中NF-κBp65和Notch1蛋白的表达。这些结果表明，CSE通过Notch1/NF-κB通路抑制ox-LDL诱导的巨噬细胞损伤。
