Gastroenterology department, Children's Hospital of Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, China.
Department of Zoology, Faculty of Science, Aswan University, Aswan, Egypt.
Expert Opin Ther Targets. 2021 Jul;25(7):597-612. doi: 10.1080/14728222.2021.1952986. Epub 2021 Aug 6.
Reactive oxygen species (ROS)-mediated inflammation plays a crucial role in ischemic brain injury. Therefore, the activation of the nuclear erythroid 2 related protein and heme-oxygenase-1 (Nrf2/HO-1) pathway by thymoquinone (TQ) could ameliorate ischemic brain damage. The photo-thrombotic method was employed to assess the impact of TQ in attenuating ischemic brain damage in C57BL/6 J mice and thy1-YFP-16 transgenic mice. In vitro study of TQ efficiency to attenuate the oxygen-glucose deprivation/reoxygenation (OGD/R) induced cell death by fluorescence-activated cell sorting (FACs) analysis was also analyzed. The protein expression levels of Nrf2/HO-1, inflammatory, and apoptotic were evaluated by immunofluorescence and western blot techniques. Besides, mRNA expression level of inducible nitric oxide synthase (iNOS), proto-oncogene (c-MYC), proto-oncogene (c-FOS), 5-hydroxytryptamine receptors (5-HT), and autophagy-related 5 (Atg5) were evaluated by RT-qPCR. The dendritic spine density of YFP slices was determined by confocal microscope. Our in vivo and in vitro results indicated that TQ significantly mitigates brain damage and motor dysfunction after ischemic stroke. These observations coincided with curtailed cell death, inflammation, oxidative stress, apoptosis, and autophagy. Most importantly, Nrf2/HO-1 signaling pathway activation by TQ was vital in the modulation of the above processes. Lastly, we found TQ to have minimal toxicity in liver tissue. Our study gives credence to TQ as a promising intervention therapy for cerebral ischemia that decreases inflammation, oxidative stress, and neuronal cell death via the Nrf2/HO-1 pathway, along with modulation of apoptotic and autophagic processes.
活性氧（ROS）介导的炎症在缺血性脑损伤中起着至关重要的作用。因此，通过百里醌（TQ）激活核红细胞 2 相关蛋白和血红素加氧酶-1（Nrf2/HO-1）途径可以改善缺血性脑损伤。采用光血栓形成方法评估 TQ 减轻 C57BL/6 J 小鼠和 thy1-YFP-16 转基因小鼠缺血性脑损伤的影响。还通过荧光激活细胞分选（FACs）分析研究了 TQ 降低氧葡萄糖剥夺/再氧合（OGD/R）诱导的细胞死亡的体外效率。通过免疫荧光和western blot 技术评估了 Nrf2/HO-1、炎症和凋亡的蛋白表达水平。此外，通过 RT-qPCR 评估了诱导型一氧化氮合酶（iNOS）、原癌基因（c-MYC）、原癌基因（c-FOS）、5-羟色胺受体（5-HT）和自噬相关 5（Atg5）的 mRNA 表达水平。通过共聚焦显微镜确定 YFP 切片的树突棘密度。我们的体内和体外结果表明，TQ 可显著减轻缺血性中风后的脑损伤和运动功能障碍。这些观察结果与细胞死亡、炎症、氧化应激、凋亡和自噬减少相一致。最重要的是，TQ 通过 Nrf2/HO-1 信号通路的激活对上述过程的调节至关重要。最后，我们发现 TQ 对肝组织的毒性最小。我们的研究为 TQ 作为一种有前途的治疗缺血性脑损伤的干预疗法提供了依据，该疗法通过 Nrf2/HO-1 途径降低炎症、氧化应激和神经元细胞死亡，同时调节凋亡和自噬过程。
