Key Laboratory of Bioresource Research and Development of Liaoning Province, College of Life and Health Sciences, Northeastern University, Shenyang, China; National Frontiers Science Center for Industrial Intelligence and Systems Optimization, Key Laboratory of Data Analytics and Optimization for Smart Industry, Ministry of Education, Northeastern University, Shenyang, China.
Key Laboratory of Bioresource Research and Development of Liaoning Province, College of Life and Health Sciences, Northeastern University, Shenyang, China.
Phytomedicine. 2023 Nov;120:155044. doi: 10.1016/j.phymed.2023.155044. Epub 2023 Aug 21.
The urgent challenge for ischemic stroke treatment is the lack of effective neuroprotectants that target multiple pathological processes. Crebanine, an isoquinoline-like alkaloid with superior pharmacological activities, presents itself as a promising candidate for neuroprotection. However, its effects and mechanisms on ischemic stroke remain unknown.
The effects of crebanine on brain damage following ischemic stroke were evaluated using the middle cerebral artery occlusion and reperfusion (MCAO/R) model. Mechanism of action was investigated using both MCAO/R rats and lipopolysaccharide (LPS)-activated BV-2 cells.
We initially demonstrated that crebanine effectively ameliorated the neurological deficits in MCAO/R rats, while also reducing brain edema and infarction. Treatment with crebanine resulted in the up-regulation of NeuN fluorescence density and down-regulation of FJB cell count, and mitigated synaptic damage. Crebanine attenuated the hyperactivation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX2) by downregulating NADP+ and NADPH levels, suppressing gp91 and p47 expressions, and reducing p47 membrane translocation in Iba-1 cells. Additionally, crebanine reduced the quantity of Iba-1 cells and protein expression. Correlation analysis has demonstrated that the inhibition of NOX2 activation in microglia is beneficial for mitigating I/R brain injuries. Moreover, crebanine exhibited significant antioxidant properties by down-regulating the expression of superoxide anion and intracellular reactive oxygen species in vivo and in vitro, and reducing lipid and DNA peroxidation. Crebanine exerted anti-inflammatory effect, as evidenced by the reduction in the expressions of nitric oxide, interleukin 1β, tumor necrosis factor α, interleukin 6, and inducible nitric oxide synthase. The effect of crebanine was achieved through the suppression of nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPK) signaling pathway. This is supported by evidence showing reduced NF-κB p65 promoter activity and nucleus translocation, as well as suppressed IκBα phosphorylation and degradation. Additionally, it inhibited the phosphorylation of ERK, JNK, and p38 MAPKs. Importantly, the anti-oxidative stress and neuroinflammation effects of crebanine were further enhanced after silencing gp91 and p47.
Crebanine alleviated the brain damages of MCAO/R rats by inhibiting oxidative stress and neuroinflammation mediated by NOX2 in microglia, implying crebanine might be a potential natural drug for the treatment of cerebral ischemia.
缺血性脑卒中治疗的当务之急是缺乏针对多种病理过程的有效神经保护剂。具有优异药理活性的异喹啉类生物碱船形乌头碱，作为神经保护的候选药物具有很大的潜力。然而，其对缺血性脑卒中的作用及机制仍不清楚。
采用大脑中动脉闭塞再灌注（MCAO/R）模型评价船形乌头碱对缺血性脑卒中后脑损伤的影响。采用 MCAO/R 大鼠和脂多糖（LPS）激活的 BV-2 细胞探讨其作用机制。
我们首先发现船形乌头碱能有效改善 MCAO/R 大鼠的神经功能缺损，减轻脑水肿和梗死体积。船形乌头碱治疗后可上调 NeuN 荧光密度，下调 FJB 细胞计数，减轻突触损伤。船形乌头碱通过下调 NADP+和 NADPH 水平、抑制 gp91 和 p47 表达以及减少 Iba-1 细胞中 p47 的膜转位，抑制烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶 2（NOX2）的过度激活。此外，船形乌头碱还可减少 Iba-1 细胞数量和蛋白表达。相关性分析表明，抑制小胶质细胞中 NOX2 的激活有利于减轻 I/R 脑损伤。此外，船形乌头碱在体内和体外均通过下调超氧阴离子和细胞内活性氧的表达以及减少脂质和 DNA 过氧化，表现出显著的抗氧化特性。船形乌头碱还具有抗炎作用，表现为降低一氧化氮、白细胞介素 1β、肿瘤坏死因子α、白细胞介素 6 和诱导型一氧化氮合酶的表达。船形乌头碱的作用是通过抑制核因子-κB（NF-κB）和丝裂原活化蛋白激酶（MAPK）信号通路来实现的。这一点可以从 NF-κB p65 启动子活性和核转位减少、IκBα 磷酸化和降解抑制以及 ERK、JNK 和 p38 MAPK 磷酸化抑制得到证明。此外，它还抑制了 gp91 和 p47 的磷酸化。重要的是，沉默 gp91 和 p47 后，船形乌头碱的抗氧化应激和神经炎症作用进一步增强。
船形乌头碱通过抑制小胶质细胞中 NOX2 介导的氧化应激和神经炎症减轻 MCAO/R 大鼠的脑损伤，提示船形乌头碱可能是治疗脑缺血的一种有潜力的天然药物。
