Zhilong Huoxue Tongyu Capsules Ameliorate Early Brain Inflammatory Injury Induced by Intracerebral Hemorrhage via Inhibition of Canonical NFκβ Signalling Pathway

Background: Intracerebral hemorrhage (ICH) is a debilitating and fatal condition with continuously rising incidence globally, without effective treatment available. Zhilong Huoxue Tongyu (ZLHXTY) capsule is a traditional Chinese medicine that is used for ICH treatment in China. However, the evidence based mechanism is not clear. Purpose: To study the protective effects of ZLHXTY capsules against ICH pathogenesis via targetting nuclear factor kappa beta (NFk beta) canonical signalling pathway. Methods: C57BL/6 J mice ICH models using autologous blood injection were used to study the effect of ZLHXTY (1.4 g/kg P.O.) after 24 and 72 hrs of ICH induction. The neurological scoring, corner turn test and balance beam with scoring was performed to assess neurological damage. Hematoxylin/eosin and nissl staining was used for histopathological evaluation. Levels of TNF alpha, NFkB, iNOS, COX2, IL1, IL6 were measured using real time qPCR and western blotting. Protein levels of IKK beta and IkB alpha were analyzed through western blotting. Immunofluorescence for co-expression of NeuN/TNF alpha, NeuN/NFkB, Iba1/TNF alpha, and Iba1/NFkB was also performed. Results: Treatment with ZLHXTY capsules after ICH ameliorated inflammatory brain injury after 24 and 72 h; revealed by neurological scoring, hematoxylin/eosin and nissl staining. The qPCR and western blot analyses demonstrated significant downregulation of TNF alpha, NFkB, iNOS, COX2, IL1 beta and IL6. Further, the IKK beta and IkB alpha revealed significant downregulation and upregulation respectively in western blot. Immunofluorescence also revealed attenuated expression of TNF alpha and NFkB in neurons and also low expression of Iba1. Conclusion: ZLHXTY capsules elicit its neuroprotective effect by targetting the NFk beta canonical signalling pathway, thereby ameliorating the ICH induced brain injury.

Automated summary

This study investigates the protective effects of ZLHXTY capsules against intracerebral hemorrhage (ICH) by targeting the nuclear factor kappa beta (NFk beta) signaling pathway. The results show that ZLHXTY capsules alleviate inflammatory brain injury after ICH induction, as evidenced by neurological scoring and histopathological evaluation. The downregulation of pro-inflammatory factors and the modulation of NFk beta pathway are associated with its neuroprotective effects.