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-1β, and TXNIP, and protein quantities of NLRP3, IL-1β, 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-1α, FoxO1, and p-FoxO1, and the co-localization of p-STAT3 with HIF-1α 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-1β, 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-1α and FoxO1 were decreased in the C + P group. While C + P inhibited HIF-1α 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-1α 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-1α/FoxO1 are vital regulators and potential targets for efficacious therapy following ischemic stroke.

据报道，氯丙嗪和异丙嗪 (C + P) 对大脑活动的抑制或冬眠样作用可诱导神经保护，无论是否诱导低温。然而，潜在的机制仍不清楚。本研究评估了 C+P 对缺血/再灌注后抑制神经炎症反应和炎症小体活化的药理作用。共有 72 只成年雄性 Sprague-Dawley 大鼠接受 2 小时大脑中动脉闭塞（MCAO），然后进行 6 或 24 小时再灌注。在再灌注开始时，大鼠接受温度控制的 C + P (8 mg/kg)。通过测量 CD68 和髓过氧化物酶 (MPO) 水平检测脑细胞死亡。通过 NLRP3、IL-1β 和 TXNIP 的 mRNA 水平以及 NLRP3、IL-1β、TXNIP、cleaved-Caspase-1、和 IL-18。通过 STAT3 (p-STAT3) 和 JAK2 (p-JAK2) 的磷酸化以及 p-STAT3 和 NLRP3 的共定位来检测 JAK2/STAT3 通路的激活。用 p-p38/p38 的蛋​​白质水平评估 p38 途径的激活。定量 HIF-1α、FoxO1 和 p-FoxO1 的 mRNA 和蛋白质水平，以及 p-STAT3 与 HIF-1α 或 FoxO1 的共定位。正如预期的那样，C + P 显着减少细胞死亡并减弱由减少的 CD68 和 MPO 确定的神经炎症反应。C + P 降低缺血诱导的炎性体活化，表现为 NLRP3、IL-1β、TXNIP、cleaved-Caspase-1 和 IL-18 的 mRNA 和蛋白质表达降低。C + P 也抑制 JAK2/STAT3 和 p38 通路的磷酸化以及 p-STAT3 与 NLRP3 的共定位。此外，C + P组中HIF-1α和FoxO1的mRNA水平降低。C+P 抑制 HIF-1α 蛋白表达的同时增加 FoxO1 的磷酸化，从而促进 FoxO1 从细胞核中排除并抑制 FoxO1 的活性。同时，C + P 减少了 p-STAT3 与 HIF-1α 或 FoxO1 的共定位。总之，C+P 治疗通过抑制神经炎症和 NLRP3 炎性体激活而赋予中风神经保护作用。本研究表明 JAK2/STAT3/p38/HIF-1α/FoxO1 是缺血性卒中后有效治疗的重要调节因子和潜在靶点。C + P 减少了 p-STAT3 与 HIF-1α 或 FoxO1 的共定位。总之，C+P 治疗通过抑制神经炎症和 NLRP3 炎性体激活而赋予中风神经保护作用。本研究表明 JAK2/STAT3/p38/HIF-1α/FoxO1 是缺血性卒中后有效治疗的重要调节因子和潜在靶点。C + P 减少了 p-STAT3 与 HIF-1α 或 FoxO1 的共定位。总之，C+P 治疗通过抑制神经炎症和 NLRP3 炎性体激活而赋予中风神经保护作用。本研究表明 JAK2/STAT3/p38/HIF-1α/FoxO1 是缺血性卒中后有效治疗的重要调节因子和潜在靶点。