Yue Xin, Wu Minghui, Jiang Hong, Hao Jing, Zhao Qinghao, Zhu Qing, Saren Gaowa, Zhang Yun, Zhang Xiaoli
Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Department of Cardiology, Qilu Hospital, Shandong University, Jinan, Shandong 250012, P.R. China.
Key Laboratory of The Ministry of Education for Experimental Teratology, Department of Histology & Embryology, Medical College, Shandong University, Jinan, Shandong 250012, P.R. China.
Mol Med Rep. 2016 Sep;14(3):1979-85. doi: 10.3892/mmr.2016.5457. Epub 2016 Jun 30.
To investigate the effects of inflammatory factor interleukin (IL)‑6 on the expression of endothelial lipase (EL) and its potential signaling pathways in atherosclerosis, a primary culture of human umbilical vein endothelial cells (HUVECs) was established and treated as follows: i) Control group without any treatment; ii) recombinant human (rh)IL‑6 treatment (10 ng/ml) for 0, 4, 8, 12 and 24 h; iii) p38 mitogen‑activated protein kinases (MAPKs) inhibitor (SB203580, 10 µmol/l) pretreatment for 1 h prior to rhIL‑6 (10 ng/ml) treatment; iv) nuclear factor (NF)‑κB activation inhibitor (pyrrolidine dithiocarbamate, 10 mmol/l) pretreatment for 1 h prior to rhIL‑6 (10 ng/ml) treatment. EL levels were detected by immunocytochemical staining and western blot analysis. Proliferation of HUVECs was detected by immunostaining of proliferating cell nuclear antigen (PCNA) and an MTT assay. p38 MAPK and NF‑κB p65 levels were detected by western blotting. The results showed that rhIL‑6 treatment increased EL expression and proliferation of HUVECs. NF‑κB p65 and MAPK p38 protein levels also increased in a time‑dependent manner in HUVECs after rhIL‑6 treatment. NF‑κB inhibitor and MAPK p38 inhibitor prevented the effects of rhIL‑6 on EL expression. In conclusion, inflammatory factor IL‑6 may participate in the pathogenesis of atherosclerosis by increasing EL expression and the proliferation of endothelial cells via the p38 MAPK and NF-κB signaling pathways.
为研究炎症因子白细胞介素（IL）-6对动脉粥样硬化中内皮脂肪酶（EL）表达及其潜在信号通路的影响，建立人脐静脉内皮细胞（HUVECs）原代培养并进行如下处理：i）对照组不做任何处理；ii）重组人（rh）IL-6处理（10 ng/ml）0、4、8、12和24小时；iii）p38丝裂原活化蛋白激酶（MAPKs）抑制剂（SB203580，10 μmol/l）在rhIL-6（10 ng/ml）处理前预处理1小时；iv）核因子（NF）-κB激活抑制剂（吡咯烷二硫代氨基甲酸盐，10 mmol/l）在rhIL-6（10 ng/ml）处理前预处理1小时。通过免疫细胞化学染色和蛋白质印迹分析检测EL水平。通过增殖细胞核抗原（PCNA）免疫染色和MTT法检测HUVECs的增殖。通过蛋白质印迹检测p38 MAPK和NF-κB p65水平。结果显示，rhIL-6处理增加了HUVECs的EL表达和增殖。rhIL-6处理后，HUVECs中NF-κB p65和MAPK p38蛋白水平也呈时间依赖性增加。NF-κB抑制剂和MAPK p38抑制剂可阻止rhIL-6对EL表达的影响。总之，炎症因子IL-6可能通过p38 MAPK和NF-κB信号通路增加EL表达和内皮细胞增殖，从而参与动脉粥样硬化的发病机制。
