Department of Endocrinology, Shandong Provincial Hospital Affiliated to Shandong University, Shandong Clinical Medical Center of Endocrinology and Metabolism, Institute of Endocrinology and Metabolism; Shandong Academy of Clinical Medicine, Jinan, Shandong China.
Eur Rev Med Pharmacol Sci. 2018 Apr;22(8):2421-2431. doi: 10.26355/eurrev_201804_14835.
To investigate the role of Toll-like receptor-4 (TLR4) in the free fatty acids (FFAs) induced human umbilical vein endothelial cells (HUVECs) inflammation and to explore the underlying mechanisms.
HUVECs and HEK293 cell lines were obtained from Shanghai Type Culture Collection. Cell counting kit-8 (CCK8) and flow cytometry (FCM) were performed to examine the cell viability and apoptosis rate of HUVECs induced by FFAs treatments with or without infection of toll-like receptor-4 interference (TLR4i) adenovirus. Enzyme-linked immunosorbent assay (ELISA) was performed to evaluate the inflammatory cytokines release. Quantitative polymerase chain reaction (qPCR) and Western Blot (WB) were used to test the molecular mechanisms of inflammation.
FFAs induced inflammatory responses in HUVECs via modulating the TLR4 receptor complex. TLR4i adenovirus interference increased cell viability and decreased cell apoptosis rate. FFAs treatments significantly increased the expressions of inflammatory cytokines interleukin-6 (IL-6), interleukin-8 (IL-8), C-C motif chemokine ligand 5 (CCL5) and CXC chemokine ligand 10 (CXCL10), while TLR4i adenovirus interference significantly reduced these cytokines levels. TLR4-mediated myeloid differential protein-88 (MyD88) expression activating the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and inhabiting kappa B kinase-beta (IKK-β). TLR4i adenovirus interference decreased the expressions of these genes at both mRNA level and protein level.
TLR4 mediates FFAs induced inflammatory responses in HUVECs. TLR4 interference in HUVECs significantly reduces the inflammatory cytokines expression, decreases the cell apoptosis rate and increases cell viability.
探讨 Toll 样受体 4（TLR4）在游离脂肪酸（FFAs）诱导的人脐静脉内皮细胞（HUVECs）炎症中的作用，并探讨其潜在机制。
本研究中的 HUVECs 和 HEK293 细胞系均购自上海细胞库。通过细胞计数试剂盒-8（CCK8）和流式细胞术（FCM）检测 FFAs 处理后感染 TLR4 干扰（TLR4i）腺病毒或不感染的 HUVECs 细胞活力和凋亡率。酶联免疫吸附试验（ELISA）用于评估炎症细胞因子的释放。实时定量聚合酶链反应（qPCR）和 Western Blot（WB）用于检测炎症的分子机制。
FFAs 通过调节 TLR4 受体复合物在 HUVECs 中诱导炎症反应。TLR4i 腺病毒干扰可增加细胞活力，降低细胞凋亡率。FFAs 处理显著增加了炎症细胞因子白细胞介素-6（IL-6）、白细胞介素-8（IL-8）、C-C 基序趋化因子配体 5（CCL5）和 CXC 趋化因子配体 10（CXCL10）的表达，而 TLR4i 腺病毒干扰则显著降低了这些细胞因子的水平。TLR4 介导的髓样分化蛋白-88（MyD88）表达激活核因子 kappa-轻链增强子的 B 细胞（NF-κB）和抑制κB 激酶-β（IKK-β）。TLR4i 腺病毒干扰降低了这些基因在 mRNA 水平和蛋白水平的表达。
TLR4 介导 FFAs 诱导的 HUVECs 炎症反应。HUVECs 中 TLR4 的干扰显著降低了炎症细胞因子的表达，降低了细胞凋亡率，增加了细胞活力。
