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目的探究白细胞介素(IL)-38在动脉粥样硬化患者血清中的水平及其对氧化型低密度脂蛋白(ox-LDL)诱导的内皮细胞炎症的作用及分子机制。方法收集65例颅内外颈动脉粥样硬化患者和30例健康人的血液,ELISA检测血清中IL-38的表达。在人动脉内皮细胞(HAECs)中先转染pcDNA3.1-IL-38质粒过表达IL-38,然后用100μg/ml ox-LDL刺激。ELISA检测细胞上清液中IL-6、IL-8、IL-17、ICAM-1和MCP-1等炎性介质的表达;real-time PCR检测细胞中凝集素样氧化低密度脂蛋白受体-1(LOX-1)和CD36 mRNA水平的表达,Western blot检测细胞中LOX-1、CD36、磷酸化核内转录因子p65(p-NF-κB p65,p-p65)和IκB蛋白水平的表达。最后转染pcDNA3.1-IL-38质粒24 h后,用100μmol/L PDTC(NF-κB抑制剂)预处理HAECs2 h,再加入100μg/ml ox-LDL刺激,检测细胞中LOX-1、IL-6、IL-8、IL-17、ICAM-1和MCP-1的表达。结果与健康人相比,脑动脉粥样硬化患者血清中IL-38的表达明显增加(P<0.05),且与其动脉粥样硬化程度呈正相关性。IL-38能够降低ox-LDL诱导的IL-6、IL-8、IL-17、ICAM-1和MCP-1等炎性介质、LOX-1 mRNA和蛋白水平、p-p65的表达,增加IκB蛋白水平表达(P均<0.05),但是对CD36 mRNA和蛋白表达无影响。PDTC能够减弱IL-38对LOX-1、IL-6、IL-8、IL-17、ICAM-1和MCP-1表达的影响。结论 IL-38的表达水平与动脉粥样硬化程度呈正相关,它能够抑制ox-LDL诱导的内皮细胞炎症,这可能与其抑制NF-κB信号通路相关。
Objective To investigate the level of interleukin-38 (IL-38) in serum of patients with atherosclerosis and its effect on endothelial cell inflammation induced by oxidized low density lipoprotein (ox-LDL) and its molecular mechanism. Methods Blood was collected from 65 patients with extracranial carotid atherosclerosis and 30 healthy individuals. The serum level of IL-38 was detected by ELISA. PcDNA3.1-IL-38 plasmids were first overexpressed in human arterial endothelial cells (HAECs) and then stimulated with 100 μg / ml ox-LDL. The expression of IL-6, IL-8, IL-17, ICAM-1 and MCP-1 in the supernatant was detected by ELISA. The expression of lectin-like oxidized low density lipoprotein receptor The expression of LOX-1, CD36 and phosphorylated nuclear transcription factor p65 (NF-κB p65, p-p65) and IκB protein were detected by Western blot . After being transfected with pcDNA3.1-IL-38 plasmid for 24 h, HAECs were pretreated with 100 μmol / L PDTC (NF-κB inhibitor) for 2 h and then stimulated with 100 μg / ml ox-LDL to detect the expression of LOX-1, IL -6, IL-8, IL-17, ICAM-1 and MCP-1. Results The serum level of IL-38 in patients with cerebral atherosclerosis was significantly higher than that in healthy people (P <0.05), and positively correlated with the level of atherosclerosis. IL-38 decreased the expression of LOX-1 mRNA and protein, p-p65, and increased IκB in the inflammatory mediators such as IL-6, IL-8, IL-17, ICAM-1 and MCP- (All P <0.05), but had no effect on the expression of CD36 mRNA and protein. PDTC attenuated the effects of IL-38 on the expression of LOX-1, IL-6, IL-8, IL-17, ICAM-1 and MCP- Conclusion The expression of IL-38 is positively correlated with the degree of atherosclerosis. It can inhibit the ox-LDL-induced endothelial cell inflammation, which may be related to the inhibition of NF-κB signaling pathway.