目的 研究视黄酸(RA)对脂多糖(LPS)诱导的血管炎症与氧化应激的保护作用并探讨其作用机制。 方法 SD大鼠给予RA和TLR4抑制剂连续口腔灌胃2周后，除对照组外，LPS组、RA 3 mg/kg组、RA 15 mg/kg组和TLR4抑制剂组（TAK-242, 3 mg/kg），通过腹腔注射LPS (10 mg/kg)以建立血管炎症模型。血管张力测定系统检测血管舒张功能，硝酸还原酶法检测大鼠血清中一氧化氮（NO）的含量，ELISA法检测血清中IL-18、IL-1β、TNF-α、IL-6和GSH-px的水平，WST-1法和TBA法分别检测血清中超氧化物歧化酶（SOD）活力、丙二醛（MDA），DHE荧光探针检测血管活性氧(ROS)的水平，免疫组化法检测血管 NF-κB p65的表达，免疫印迹法检测血管TLR4、eNOS和p-eNOS的表达。 结果 与LPS组相比，RA能够改善血管舒张功能，增加p-eNOS和NO的水平，减少血清炎症因子IL-18、IL-1β、TNF-α、IL-6的含量，降低血清中MDA和ROS的生成，升高血清中SOD的生成和GSH-px的释放量，并且下调血管TLR4和NF-κB p65的表达水平。另外，RA对LPS诱导的血管炎症与氧化应激的影响与TLR4抑制剂作用相似。 结论 RA对LPS诱导的血管炎症与氧化应激具有抑制作用，其可能通过TLR4/NF-κB p65信号通路发挥作用。
Abstract
Objective To study potential protective effects of retinoic acid (RA) on lipopolysaccharide (LPS)-induced vascular inflammation and oxidative stress and explore its possible molecular mechanism. Methods After SD rats were given RA and TLR4 inhibitors by oral gavage for 2 weeks, except for the control group, LPS group, RA 3 mg/kg group, RA 15 mg/kg group and TLR4 inhibitor group (TAK-242, 3 mg/kg), LPS (10 mg/kg) was injected intraperitoneally to establish a vascular inflammation model. Vascular tension measurement system was used to detect vasodilation function, nitrate reductase method was used to detect nitric oxide (NO) content in rat serum. Serum IL-18, IL-1β, TNF-α, IL-6 and GSH- px level was detected by ELISA method, WST-1 and TBA methods were used to detect the activity of superoxide dismutase (SOD) and malondialdehyde (MDA) in the serum respectively, DHE fluorescent probe was used to detect the level of vascular reactive oxygen species (ROS), and immunohistochemical method to detect the expression of vascular NF-κB p65, Western blotting to detect the expression of vascular TLR4, eNOS and p-eNOS. Results Compared with the LPS group, RA can improve the vasodilation function, increase the levels of p-eNOS and NO, decrease the serum inflammatory factors IL-18, IL-1β, TNF-α, IL-6 level, and reduce the serum MDA and ROS, increase the production of SOD and the release of GSH-px in the serum, and down-regulate the expression levels of vascular TLR4 and NF-κB p65. In addition, the effect of RA on LPS-induced vascular inflammation and oxidative stress was similar to that of TLR4 inhibitors. Conclusions RA has an inhibitory effect on LPS-induced vascular inflammation and oxidative stress, through the TLR4/NF-κB p65 signaling pathway.
关键词
视黄酸； /
/
TLR4/NF-κB； /
/
血管内皮功能障碍； /
/
炎症； /
/
氧化应激
Key words
Retinoic acid; /
/
TLR4/NF-κB; /
Vascular endothelial dysfunction; /
Inflammation; /
Oxidative stress
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基金
辽宁省兴辽计划领军人才专项资金（XLYC 1802017）
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