Therapeutic impact of thymoquninone to alleviate ischemic brain injury via Nrf2/HO-1 pathway

ABSTRACT

Introduction: 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.

Areas covered: 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.

Results: 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.

Conclusion: 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.

KEYWORDS:

• Cerebral ischemia
• thymoquinone
• nuclear factor erythroid 2-related heme oxygenase–1
• oxidative stress
• oxygen-glucose deprivation/reoxygenation

Availability of data and materials

The datasets during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Declaration of interest

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Author contributions

Marong Fang and Zhiying Hu designed the experiments. Nashwa Amin performed the experiment, drafted the manuscript, and analyzed the data. Qiannan Ren participated in the in vitro study. Xiaoxue Du, Shijia Chen, Benson O.A. Botchway, and Azhar B. Hussien are mainly responsible for polishing the article and manuscript editing and writing. Marong Fang is mainly responsible for obtained funding. All authors read and approved the final manuscript.

Reviewer disclosures

Peer reviewers on this manuscript have no relevant financial or other relationships to disclose

Supplementary Material

Supplemental data for this article can be accessed here.

Additional information

Funding

The National Natural Science Foundation of China (grant numbers 81971769 and 81871063) supported this study. We extend our appreciation to the Core Facilities of Zhejiang University Institute of Neuroscience for their technical assistance, facilities, and support throughout this study.