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(1 μgmL-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(1 μgmL-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(1 μgmL-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(5,10,20 μmolL-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。
Abstract
OBJECTIVE To investigate the inhibitory effects of caffeic acid phenethyl ester (CAPE) on LPS-induced inflammatory responses in BV-2 microglial cells and its mechanism.METHODS The model was build by BV-2 microglial cells induced by LPS (1 μgmL-1), which was used to inspect the cell survivability by means of MTT, investigate the effect of CAPE on the release of inflammatory factor NO, IL-1β, IL-6 by ELISA, and the expression of inflammation-related protein (iNOS, COX-2, IκB, P-IκB, NF-κB, P-NF-κB) were analyzed by used of Western Blot. Finally, the regulatory effects of CAPE on inflammatory transcription factor NF-κB activation were inspected. RESULTS CAPE and LPS affected on BV-2 cells, which are collected after being incubated at 37 ℃ for 24 or 8 h, then test the release of NO, IL-1β, IL-6. As is shown, LPS (1 μgmL-1) can injury the cells,and increase inflammatory cytokines. CAPE (5, 10, 20 μmolL-1) can decrease these inflammatory cytokines, down-regulate iNOS, COX-2, IκB, P-IκB expression and restrain NF-κB nuclear translocation. CONCLUSION The anti-inflammatory activity of CAPE may be achieved through inhibiting NF-κB signaling pathway.
关键词
小胶质细胞 /
神经炎症 /
咖啡酸苯乙酯 /
抗炎 /
脂多糖
{{custom_keyword}} /
Key words
BV-2 microglia cell /
neuroinflammation /
caffeic acid phenethyl ester /
anti-inflammation /
lipopolysaccharide
{{custom_keyword}} /
中图分类号：
R965
{{custom_clc.code}}
({{custom_clc.text}})
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] HUANG Y, MUCKE L. Alzheimer mechanisms and therapeutic strategies [J]. Cell, 2012, 148(6): 1204-1222.[2] ZHANG W, LEE L, CHU Y Q, et al. Effects of ginsenoside on nerve cells injuried by Aβ-induced monocyte supernatant [J]. Chin Pharm J (中国药学杂志), 2007, 42(21): 1626-1628.[3] XUE Y, LEE H F, XU L Y. Microglia: In inflammation-mediated neurodegenerative diseases[J]. Chin Bull Life Sci (生命科学), 2007, 19(1): 43-46.[4] TOMOHARU T, SHINICHI K, TOMONORI M, et al.General anesthetics inhibit LPS-induced IL-1β expression in glial cells [J]. Plos One, 2013, 8(12): 1-13.[5] ZHAO W X, WANG L, YANG J L, et al. Caffeic acid phenethyl ester attenuates pro-inflammatory and fibrogenic phenotypes of LPS-stimulated hepatic stellate cells through the inhibition of NF-kappaB signaling [J]. Int J Mol Med, 2014, 33(3): 687-694.[6] KOKSEL O, OZDULGER A, TAMER L, et al. Effects of caffeic acid phenethyl ester on lipopolysaccharide-induced lung injury in rats [J]. Pulm Pharmacol Ther, 2006, 19(2): 90-95.[7] LI Y, DU X F, DU J L. Physiological properties and functions of microglia [J]. Acta Physiol Sin(生理学报), 2013, 65(5): 471-482.[8] PERRY V H, NICOLL J A, HOLMES C. Microglia in neurodegenerative disease [J]. Nat Rev Neurol, 2010, 6(4): 193-201.[9] KRAUSE D L, MULLER N. Neuroinflammation, microglia and implications for anti-inflammatory treatment in Alzheimer's disease [J]. Int J Alzheimers Dis, 2010,2010. pii: 732806. doi: 10.4061/2010/732806.[10] BANNO M, MIZUNO T, KATO H, et al. The radical scavenger edaravone prevents oxidative neurotoxicity induced by peroxynitrite and activated microglia [J]. Neuropharmacology, 2005, 48(2): 283-290.[11] YU S C, LI X Y. Effect of ginsenoside on IL-1 beta and IL-6 mRNA expression in hippocampal neurons in chronic inflammation model of aged rats [J]. Acta Pharmacol Sin(中国药理学报), 2000, 21(10): 915-918.[12] KARIN M, YAMAMOTO Y, WANG Q M. The IKK NF-kappa B system: A treasure trove for drug development [J]. Nat Rev Drug Discov, 2004, 3(1): 17-26.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
基金
国家自然科学基金资助项目(30873072)
{{custom_fund}}
PDF(1553 KB)
