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目的探究咖啡酸苯乙酯对脂多糖诱导的小胶质细胞BV-2神经炎症反应的抑制作用及其潜在的作用机制。方法采用脂多糖(1μg·mL-1)诱导小胶质细胞BV-2活化,制作炎症反应模型,利用四甲基偶氮唑蓝法检测细胞存活率,并用酶联免疫法测定咖啡酸苯乙酯对炎症因子一氧化氮、白细胞介素-1β和白细胞介素-6合成释放的影响。然后用蛋白印迹法研究了咖啡酸苯乙酯对炎症相关蛋白(诱导型一氧化氮合酶、环氧化酶-2、IκB以及P-IκB)表达的作用。此外,进一步研究了咖啡酸苯乙酯对炎症转录因子NF-κB核转位的作用。结果咖啡酸苯乙酯与脂多糖(1μg·mL-1)同时作用于BV-2小胶质细胞,分别于温箱中孵育24、8 h收集上清液,检测一氧化氮、白细胞介素-1β、白细胞介素-6的释放量。结果显示,脂多糖(1μg·mL-1)可以造成细胞损伤,增加炎症因子释放,咖啡酸苯乙酯则可以降低脂多糖诱导的小胶质细胞对一氧化氮、白细胞介素-1β和白细胞介素-6的释放,并可抑制诱导型一氧化氮合酶、环氧化酶-2、P-IκB、P-NF-κB蛋白的表达,还可以抑制NF-κB的核转位。结论上述研究说明,咖啡酸苯乙酯(5,10,20μmol·L-1)可以通过抑制炎性因子的表达而减少对小胶质细胞的损伤,其抗炎活性可能是通过抑制NF-κB信号通路介导的炎症反应实现的。
Objective To investigate the inhibitory effect of caffeic acid phenethyl ester on lipopolysaccharide-induced microglia BV-2 neuroinflammation and its potential mechanism. Methods Lipopolysaccharide (1μg · mL-1) was used to induce activation of BV-2 in microglia. The inflammatory response model was established. Cell viability was measured by MTT method and the content of caffeic acid Effects of Ester on the Synthesis and Release of Inflammatory Factors Nitric Oxide, Interleukin - 1β and Interleukin - 6. The effect of caffeic acid phenethyl ester on the expression of inflammatory-associated proteins (inducible nitric oxide synthase, cyclooxygenase-2, IκB, and P-IκB) was then studied by Western blotting. In addition, the effect of caffeic acid phenethyl ester on nuclear translocation of inflammatory transcription factor NF-κB was further studied. Results Caffeic acid phenethyl ester and lipopolysaccharide (1μg · mL-1) were applied to BV-2 microglial cells at the same time. The supernatant was collected for 24 and 8 hours respectively in incubators. Nitric oxide, interleukin -1β, interleukin-6 release. The results showed that lipopolysaccharide (1μg · mL-1) could cause cell injury and increase the release of inflammatory cytokines, and caffeic acid phenethyl ester could reduce the effect of lipopolysaccharide-induced microglial on nitric oxide, interleukin-1β and leukocyte 6 release, and can inhibit the expression of inducible nitric oxide synthase, cyclooxygenase-2, P-IκB, P-NF-κB protein, but also can inhibit the nuclear translocation of NF-κB. Conclusion The above study shows that caffeic acid phenethyl ester (5,10,20μmol·L-1) can reduce the damage to microglia by inhibiting the expression of inflammatory cytokines, and its anti-inflammatory activity may be through inhibition of NF-κB Signal Pathway-mediated Inflammation Response Achieved.