Huang-Qi-Long-Dan Granule alleviates ischemic stroke injury by regulating the crosstalk between Nrf2 and NF-κB signaling

Background: Huang-Qi-Long-Dan Granule (HQLDG) is an herbal formula that is commonly used to treat ischemic stroke (IS) in clinical practice. Comprehension of the underlying mechanism is conductive to an improved understanding of the use of this formula.

Purpose: The anti-IS effect and the underlying mechanism of HQLDG were explored in this study.

Methods: A zebrafish cerebral thrombosis model and a mouse transient middle cerebral artery occlusion (tMCAO) model were used in this study.

Results: HQLDG significantly decreased brain thrombosis; increased blood vessel diameter and blood flow velocity; ameliorated motor activity, including velocity, acceleration, sinuosity, etc.; and regulated expression of the coagulation cascade genes vWF, proca and tfpia in zebrafish model. Ameliorated motor activity and improved brain pathology were also observed in tMCAO mice. Furthermore, HQLDG decreased oxidative stress-, inflammation- and apoptosis- related indicators in zebrafish, which were also confirmed in tMCAO mice. Moreover, HQLDG could increase the translocation of Nrf2 into the nucleus in brain tissue. However, the protective effects of HQLDG on cerebral thrombosis zebrafish were decreased after the inhibition of Nrf2 with ML385 or CRISPR/Cas9 gene-editing technology in zebrafish. Moreover, inhibition of Nrf2 significantly attenuated the effect of HQLDG on NF-κB. The protective effect of HQLDG on motor activity was significantly decreased in NF-κB inhibited gene-edited zebrafish.

Conclusions: These findings underscore the potential of HQLDG as a promising therapeutic agent for IS, while further confirming the reliability of zebrafish as an IS model and expanding the visualization indicators of IS to complement mammalian indicators.

Huang-Qi-Long-Dan Granule; Ischemic stroke; Nuclear factor erythroid 2-related factor 2; Nuclear factor kappa-B, CRISPR/Cas9.