Phenothiazine Inhibits Neuroinflammation and Inflammasome Activation Independent of Hypothermia After Ischemic Stroke
Phenothiazine Inhibits Neuroinflammation and Inflammasome Activation Independent of Hypothermia After Ischemic Stroke
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DOI：
10.1007/s12035-021-02542-3
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发表时间：
2021-08-29
影响因子：
5.1
通讯作者：
Ding, Yuchuan
中科院分区：
文献类型：
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作者：
Guo, Sichao;Geng, Xiaokun;Ding, Yuchuan
据报道，氯丙嗪和异丙嗪（C + P）对脑活动的抑制或冬眠样作用可诱导神经保护，伴或不伴诱导低温。然而，其潜在机制仍不清楚。本研究旨在探讨C + P对缺血再灌注后神经炎症反应和炎性小体激活的抑制作用。共72只成年雄性SD大鼠进行大脑中动脉闭塞（MCAO）2小时，然后再灌注6或24小时。在再灌注开始时，大鼠接受C + P（8 mg/kg）并控制温度。通过测量CD 68和髓过氧化物酶（MPO）水平检测脑细胞死亡。通过NLRP 3、IL-1 β和TXNIP的mRNA水平以及NLRP 3、IL-1 β、TXNIP、切割的半胱天冬酶-1和IL-18的蛋白质量来测量炎性体活化。通过STAT 3（p-STAT 3）和JAK 2（p-JAK 2）的磷酸化以及p-STAT 3和NLRP 3的共定位来检测JAK 2/STAT 3通路的激活。用p-p38/p38蛋白水平评估p38通路的激活。定量HIF-1 α、FoxO 1和p-FoxO 1的mRNA和蛋白水平，以及p-STAT 3与HIF-1 α或FoxO 1的共定位。正如预期的那样，C + P显著减少了细胞死亡，并减弱了神经炎症反应，如通过减少CD 68和MPO所确定的。C + P降低了缺血诱导的炎性小体活化，表现为NLRP 3、IL-1 β、TXNIP、裂解型Caspase-1和IL-18的mRNA和蛋白表达降低。C + P还抑制JAK 2/STAT 3和p38通路的磷酸化以及p-STAT 3与NLRP 3的共定位。此外，C + P组中HIF-1 α和FoxO 1的mRNA水平降低。虽然C + P抑制HIF-1 α蛋白表达，但它增加了FoxO 1磷酸化，这促进了FoxO 1从细胞核中排出并抑制FoxO 1活性。同时，C + P减少了p-STAT 3与HIF-1 α或FoxO 1的共定位。总之，C + P治疗通过抑制神经炎症和NLRP 3炎性体激活来赋予中风中的神经保护。目前的研究表明，JAK 2/STAT 3/p38/HIF-1 α/FoxO 1是缺血性卒中后重要的调节因子和有效治疗的潜在靶点。
A depressive or hibernation-like effect of chlorpromazine and promethazine (C + P) on brain activity was reported to induce neuroprotection, with or without induced-hypothermia. However, the underlying mechanisms remain unclear. The current study evaluated the pharmacological function of C + P on the inhibition of neuroinflammatory response and inflammasome activation after ischemia/reperfusion. A total of 72 adult male Sprague-Dawley rats were subjected to 2 h middle cerebral artery occlusion (MCAO) followed by 6 or 24 h reperfusion. At the onset of reperfusion, rats received C + P (8 mg/kg) with temperature control. Brain cell death was detected by measuring CD68 and myeloperoxidase (MPO) levels. Inflammasome activation was measured by mRNA levels of NLRP3, IL-1 beta, and TXNIP, and protein quantities of NLRP3, IL-1 beta, TXNIP, cleaved-Caspase-1, and IL-18. Activation of JAK2/STAT3 pathway was detected by the phosphorylation of STAT3 (p-STAT3) and JAK2 (p-JAK2), and the co-localization of p-STAT3 and NLRP3. Activation of the p38 pathway was assessed with the protein levels of p-p38/p38. The mRNA and protein levels of HIF-1 alpha, FoxO1, and p-FoxO1, and the co-localization of p-STAT3 with HIF-1 alpha or FoxO1 were quantitated. As expected, C + P significantly reduced cell death and attenuated the neuroinflammatory response as determined by reduced CD68 and MPO. C + P decreased ischemia-induced inflammasome activation, shown by reduced mRNA and protein expressions of NLRP3, IL-1 beta, TXNIP, cleaved-Caspase-1, and IL-18. Phosphorylation of JAK2/STAT3 and p38 pathways and the co-localization of p-STAT3 with NLRP3 were also inhibited by C + P. Furthermore, mRNA levels of HIF-1 alpha and FoxO1 were decreased in the C + P group. While C + P inhibited HIF-1 alpha protein expression, it increased FoxO1 phosphorylation, which promoted the exclusion of FoxO1 from the nucleus and inhibited FoxO1 activity. At the same time, C + P reduced the co-localization of p-STAT3 with HIF-1 alpha or FoxO1. In conclusion, C + P treatment conferred neuroprotection in stroke by suppressing neuroinflammation and NLRP3 inflammasome activation. The present study suggests that JAK2/STAT3/p38/HIF-1 alpha/FoxO1 are vital regulators and potential targets for efficacious therapy following ischemic stroke.