Role of NLRP3 inflammasome in the inflammatory response induced by cortical spreading depression in mice

• Main Authors: Hsun-Ting Hsieh, 謝勛庭
• Other Authors: Yueh-Hsin Ping
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/kq452u

碩士 === 國立陽明大學 === 藥理學研究所 === 106 === Cortical spreading depression (CSD) is the pathophysiological basis of migraine aura and the trigger of migraine headaches. It is known that CSD can induce neuronal caspase-1 activation and release of interleukin-1β(IL-1β) from neurons. However, the mechanism underlying CSD-induced neuronal caspase-1 activation remains unknown. This study aimed to investigate the role of NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome in CSD-evoked cortical inflammation. In this study, C57BL/6 mice were used and anaesthetized with isoflurane. The skulls of mice were then drilled for 2 burr holes. One was the site for CSD induction, the other was the site for CSD recording. CSD was recorded by a glass microelectrode. KCl (300 mM) absorbed in a cotton ball was used to induce CSD. The result showed that the protein levels of NLRP3 in the cortex were increased by KCl-evoked CSD in a time-dependent manner. Similarly, following CSD generation induced by KCl for 2 hours, the expression levels of caspase-1 and IL-1β were significantly higher than that in the sham group. Moreover, induction of NLRP3 colocalized with NeuN was observed in the cortex after 2 hours stimulation of KCl-induced CSD. We further found that the increased protein levels of caspase-1 and IL-1β could be prevented by pretreatment with MCC950, an inhibitor of NLRP3 inflammasome activation. Also, A438079, an inhibitor of P2X7/PANX1 complex, could mildly suppress the increase of expressions of NLRP3, caspase-1 and IL-1β. In conclusion, CSD may increase caspase-1 and IL-1β production in mice cortex via NLRP3 inflammasome assembly which was triggered by activation of P2X7/PANX1 complex.