中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2014年
5期
652-656
,共5页
丹皮酚%LPS%ATP%小胶质细胞%NLRP3 炎症小体%ROS
丹皮酚%LPS%ATP%小膠質細胞%NLRP3 炎癥小體%ROS
단피분%LPS%ATP%소효질세포%NLRP3 염증소체%ROS
paeonol%LPS%ATP%microglia%NLRP3 in-flammasome%ROS
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。
目的：觀察丹皮酚( Pae)對脂多糖( LPS)與三燐痠腺苷( ATP)誘導大鼠原代小膠質細胞NLRP3炎癥小體激活的影響,探討Pae對小膠質細胞炎癥反應的抑製作用及其具體機製。方法採用白細胞分化抗原11b(CD11b)免疫熒光染色法鑒定小膠質細胞；採用ELISA法測定培養液中白細胞介素-1β( IL-1β)的水平；採用Western blot檢測細胞NLRP3、ASC和caspase-1蛋白錶達水平；採用2′,7′-二氯二氫熒光素二乙酯( DCFH-DA)為熒光探針檢測細胞內活性氧( ROS)的水平。結果 LPS(0.5 mg·L-1)/ATP(5 mmol·L-1)能增加小膠質細胞ROS及上清液IL-1β水平,上調細胞NLRP3、ASC和caspase-1蛋白水平；Pae能減少細胞 ROS和上清液IL-1β水平,抑製LPS和ATP雙信號上調的NLRP3、ASC和caspase-1蛋白水平。結論 Pae能抑製LPS/ATP激活的小膠質細胞NLRP3炎癥小體,減少細胞上清液IL-1β水平,Pae對NLRP3炎癥小體抑製作用可能與其下調小膠質細胞ROS水平有關。
목적：관찰단피분( Pae)대지다당( LPS)여삼린산선감( ATP)유도대서원대소효질세포NLRP3염증소체격활적영향,탐토Pae대소효질세포염증반응적억제작용급기구체궤제。방법채용백세포분화항원11b(CD11b)면역형광염색법감정소효질세포；채용ELISA법측정배양액중백세포개소-1β( IL-1β)적수평；채용Western blot검측세포NLRP3、ASC화caspase-1단백표체수평；채용2′,7′-이록이경형광소이을지( DCFH-DA)위형광탐침검측세포내활성양( ROS)적수평。결과 LPS(0.5 mg·L-1)/ATP(5 mmol·L-1)능증가소효질세포ROS급상청액IL-1β수평,상조세포NLRP3、ASC화caspase-1단백수평；Pae능감소세포 ROS화상청액IL-1β수평,억제LPS화ATP쌍신호상조적NLRP3、ASC화caspase-1단백수평。결론 Pae능억제LPS/ATP격활적소효질세포NLRP3염증소체,감소세포상청액IL-1β수평,Pae대NLRP3염증소체억제작용가능여기하조소효질세포ROS수평유관。
Aim To investigate the effects of paeonol on lipopolysaccharide ( LPS) and adenosine 5′-triphos-phate ( ATP) induced NLRP3 inflammasome activation in primary rat microglia and the mechanisms responsi-ble for this anti-inflammatory effects. Methods Pri-mary rat microglia were identified immunohistochemi-cally using the cluster of differentiation 11 b ( CD11 b ) antibody. Proinflammatory cytokine IL-1β was deter-mined by ELISA. Western blot was performed to ob-serve the protein expression of NLRP3 , ASC and caspase-1 in cultured primary rat microglia. The level of intracellular reactive oxygen species ( ROS) was mo-nitored by using the fluorescent probe 2′, 7′-dichlo-rofluorescein 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 signifi-cantly decreased intracellular ROS level and IL-1β se-cretion, 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 pri-mary rat microglia, and this inhibitory effect may be through the suppression of intracellular ROS.
