丹皮酚对脂多糖／三磷酸腺苷诱导的小胶质细胞NLRP3炎症小体激活的影响  

Effects of paeonol on lipopolysaccharide /adenosine 5'-triphosphate induced NLRP3 inflammasome activation in primary rat microglia

作　　者：王伟[1] 戴敏[2] 徐忠东[1] 

机构地区：[1]合肥师范学院生命科学系,安徽合肥230601 [2]安徽中医药大学药学院 安徽省中药研究与开发重点实验室 省部共建新安医学教育部重点实验室,安徽合肥230038

出　　处：《中国药理学通报》2014年第5期652-656,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81274134);安徽省自然科学基金资助项目(No 1408085MH146);安徽省高校省级自然科学研究项目(No KJ2013Z275,KJ2014A202)

摘　　要：目的观察丹皮酚(Pae)对脂多糖(LPS)与三磷酸腺苷(ATP)诱导大鼠原代小胶质细胞NLRP3炎症小体激活的影响,探讨Pae对小胶质细胞炎症反应的抑制作用及其具体机制。方法采用白细胞分化抗原11b(CD11b)免疫荧光染色法鉴定小胶质细胞;采用ELISA法测定培养液中白细胞介素-1β(IL-1β)的水平;采用Western blot检测细胞NLRP3、ASC和caspase-1蛋白表达水平;采用2',7'-二氯二氢荧光素二乙酯(DCFH-DA)为荧光探针检测细胞内活性氧(ROS)的水平。结果 LPS(0.5 mg·L-1)/ATP(5 mmol·L-1)能增加小胶质细胞ROS及上清液IL-1β水平,上调细胞NLRP3、ASC和caspase-1蛋白水平;Pae能减少细胞ROS和上清液IL-1β水平,抑制LPS和ATP双信号上调的NLRP3、ASC和caspase-1蛋白水平。结论 Pae能抑制LPS/ATP激活的小胶质细胞NLRP3炎症小体,减少细胞上清液IL-1β水平,Pae对NLRP3炎症小体抑制作用可能与其下调小胶质细胞ROS水平有关。Aim To investigate the effects of paeonol on lipopolysaccharide （LPS） and adenosine 5 ＇-triphosphate （ATP） induced NLRP3 inflammasome activation in primary rat microglia and the mechanisms responsible for this anti-inflammatory effects. Methods Primary rat microglia were identified immunohistochemically using the cluster of differentiation 11 b （CD11 b） antibody. Proinflammatory cytokine IL-1β was determined by ELISA. Western blot was performed to observe the protein expression of NLRP3, ASC and caspase-1 in cultured primary rat microglia. The level of intracellular reactive oxygen species （ROS） was monitored by using the fluorescent probe 2＇, 7＇-dichlorofluorescein diacetate （ DCFH-DA ）. Results LPS （0. 5 mg. L-1 ）/ATP （5 mmol. L-1） significantly increased intracellular ROS level and IL-1β secretion and upregulated NLRP3, ASC and caspase-1 protein expression in primary rat microglia. Paeonol significantly decreased intracellular ROS level and IL-1 β secretion, and inhibited LPS/ATP induced overexpres- sion of NLRP3, ASC and caspase-1 in cultured primary rat microglia. Conclusion Paeonol can inhibit LPS/ ATP induced NLRP3 inflammasome activation in primary rat microglia, and this inhibitory effect may be through the suppression of intracellular ROS.

关 键 词：丹皮酚 LPS ATP 小胶质细胞 NLRP3炎症小体 ROS 

分 类 号：R-332[医药卫生] R284.1