Abstract：
BACKGROUND: Saffron, a traditional Chinese medicine, is derived from Crocus sativus L. stigmas and has been reported to possess neuroprotective properties and potentially contribute to the inhibition of apoptosis and inflammation. Safranal, a potent monothyral aldehyde, is a main component of saffron that has been reported to have antiepileptic activity. However, the specific mechanism by which safranal suppresses epileptic seizures via its antiapoptotic and anti-inflammatory properties is unclear.
OBJECTIVE: To evaluate the effect of safranal on seizure severity, inflammation, and postictal neuronal apoptosis in a mouse model of pentetrazole (PTZ)-induced seizures and explore the underlying mechanism involved.
METHODS: The seizure stage and latency of stage 2 and 4 were quantified to assess the efficacy of safranal in mitigating PTZ-induced epileptic seizures in mice. Electroencephalography (EEG) was employed to monitor epileptiform afterdischarges in each experimental group. The cognitive abilities and motor functions of the mice were evaluated using the novel object recognition test and the open field test, respectively. Neurons were quantified using hematoxylin and eosin staining. Additionally, bioinformatics tools were utilized to predict the interactions between safranal and specific target proteins. Glycogen synthase kinase-3β (GSK-3β), mitochondrial apoptosis-related proteins, and inflammatory factor levels were analyzed through western blotting. Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) concentrations in brain tissue were assessed by ELISA.
RESULTS: Safranal decreased the average seizure stage and increased the lantency of stage 2 and 4 seizures in PTZ-induced epileptic mice. Additionally, safranal exhibited neuroprotective effects on hippocampal CA1 and CA3 neurons and reduced hyperactivity caused by postictal hyperexcitability. Bioinformatics analysis revealed that safranal can bind to five specific proteins, including GSK-3β. By promoting Ser9 phosphorylation and inhibiting GSK-3β activity, safranal effectively suppressed the NF-κB signaling pathway. Moreover, the findings indicate that safranal treatment can decrease TNF-α and IL-1β levels in the cerebral tissues of epileptic mice and downregulate mitochondrial apoptosis-related proteins, including Bcl-2, Bax, Bak, Caspase 9, and Caspase 3.
CONCLUSIONS: Safranal can suppress the NF-κB signaling pathway and mitochondrial-dependent apoptosis through GSK-3β inactivation, suggesting that it is a promising therapeutic agent for epilepsy treatment.
OBJECTIVE: To evaluate the effect of safranal on seizure severity, inflammation, and postictal neuronal apoptosis in a mouse model of pentetrazole (PTZ)-induced seizures and explore the underlying mechanism involved.
METHODS: The seizure stage and latency of stage 2 and 4 were quantified to assess the efficacy of safranal in mitigating PTZ-induced epileptic seizures in mice. Electroencephalography (EEG) was employed to monitor epileptiform afterdischarges in each experimental group. The cognitive abilities and motor functions of the mice were evaluated using the novel object recognition test and the open field test, respectively. Neurons were quantified using hematoxylin and eosin staining. Additionally, bioinformatics tools were utilized to predict the interactions between safranal and specific target proteins. Glycogen synthase kinase-3β (GSK-3β), mitochondrial apoptosis-related proteins, and inflammatory factor levels were analyzed through western blotting. Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) concentrations in brain tissue were assessed by ELISA.
RESULTS: Safranal decreased the average seizure stage and increased the lantency of stage 2 and 4 seizures in PTZ-induced epileptic mice. Additionally, safranal exhibited neuroprotective effects on hippocampal CA1 and CA3 neurons and reduced hyperactivity caused by postictal hyperexcitability. Bioinformatics analysis revealed that safranal can bind to five specific proteins, including GSK-3β. By promoting Ser9 phosphorylation and inhibiting GSK-3β activity, safranal effectively suppressed the NF-κB signaling pathway. Moreover, the findings indicate that safranal treatment can decrease TNF-α and IL-1β levels in the cerebral tissues of epileptic mice and downregulate mitochondrial apoptosis-related proteins, including Bcl-2, Bax, Bak, Caspase 9, and Caspase 3.
CONCLUSIONS: Safranal can suppress the NF-κB signaling pathway and mitochondrial-dependent apoptosis through GSK-3β inactivation, suggesting that it is a promising therapeutic agent for epilepsy treatment.
摘要：
背景：藏红花，中药，来源于番红花柱头，据报道具有神经保护特性，并可能有助于抑制细胞凋亡和炎症。Safranal,一种强效的单丁醛,是藏红花的主要成分，据报道具有抗癫痫活性。然而,safranal通过其抗凋亡和抗炎特性抑制癫痫发作的具体机制尚不清楚.
目的：为了评估安全治疗对癫痫发作严重程度的影响，炎症,在戊四唑（PTZ）诱导的癫痫发作小鼠模型中和后神经元凋亡，并探讨其相关机制。
方法：对第2阶段和第4阶段的癫痫发作阶段和潜伏期进行定量，以评估safranal在减轻PTZ诱导的小鼠癫痫发作中的功效。在每个实验组中，采用脑电图（EEG）监测癫痫样放电后。采用新型物体识别试验和开场试验对小鼠的认知能力和运动功能进行评价,分别。使用苏木精和曙红染色对神经元进行定量。此外,生物信息学工具被用来预测安全蛋白和特定靶蛋白之间的相互作用。糖原合成酶激酶-3β(GSK-3β),线粒体凋亡相关蛋白,通过蛋白质印迹分析炎症因子水平。ELISA法检测脑组织中肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的浓度。
结果：Safranal降低了PTZ诱导的癫痫小鼠的平均癫痫发作阶段，增加了第2阶段和第4阶段的癫痫发作。此外,safranal对海马CA1和CA3神经元具有神经保护作用，并减少了由后兴奋过度引起的过度活动。生物信息学分析显示，Safranal可以与5种特定的蛋白质结合，包括GSK-3β。通过促进Ser9磷酸化和抑制GSK-3β活性，safranal有效抑制NF-κB信号通路。此外，结果表明，三联疗法可以降低癫痫小鼠脑组织中TNF-α和IL-1β的水平，下调线粒体凋亡相关蛋白，包括Bcl-2,Bax,Bak,Caspase9和Caspase3。
结论：Safranal可通过GSK-3β失活抑制NF-κB信号通路和线粒体依赖性细胞凋亡,这表明它是一种有前途的治疗癫痫的药物。
目的：为了评估安全治疗对癫痫发作严重程度的影响，炎症,在戊四唑（PTZ）诱导的癫痫发作小鼠模型中和后神经元凋亡，并探讨其相关机制。
方法：对第2阶段和第4阶段的癫痫发作阶段和潜伏期进行定量，以评估safranal在减轻PTZ诱导的小鼠癫痫发作中的功效。在每个实验组中，采用脑电图（EEG）监测癫痫样放电后。采用新型物体识别试验和开场试验对小鼠的认知能力和运动功能进行评价,分别。使用苏木精和曙红染色对神经元进行定量。此外,生物信息学工具被用来预测安全蛋白和特定靶蛋白之间的相互作用。糖原合成酶激酶-3β(GSK-3β),线粒体凋亡相关蛋白,通过蛋白质印迹分析炎症因子水平。ELISA法检测脑组织中肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的浓度。
结果：Safranal降低了PTZ诱导的癫痫小鼠的平均癫痫发作阶段，增加了第2阶段和第4阶段的癫痫发作。此外,safranal对海马CA1和CA3神经元具有神经保护作用，并减少了由后兴奋过度引起的过度活动。生物信息学分析显示，Safranal可以与5种特定的蛋白质结合，包括GSK-3β。通过促进Ser9磷酸化和抑制GSK-3β活性，safranal有效抑制NF-κB信号通路。此外，结果表明，三联疗法可以降低癫痫小鼠脑组织中TNF-α和IL-1β的水平，下调线粒体凋亡相关蛋白，包括Bcl-2,Bax,Bak,Caspase9和Caspase3。
结论：Safranal可通过GSK-3β失活抑制NF-κB信号通路和线粒体依赖性细胞凋亡,这表明它是一种有前途的治疗癫痫的药物。
