Asiaticoside Alleviates Neuroinflammatory Complications of Intracerebral Hemorrhage in Adult Zebrafish

Background

One of the etiologies of stroke is intracerebral hemorrhage and the demand for potential therapeutic agents to alleviate symptoms and improve quality of life in patients with ICH is growing. Several phytocompounds capable of such potency with minimal adverse effects can replace drugs in market. This requires high throughput screening and in vivo experimental demonstration of the potency in ICH models that simulate mammalian pathophysiology. We aimed to study the neuroprotective activity of Asiaticoside using an adult zebrafish ICH model.

Materials and Methods

Atorvastatin-induced ICH models were treated with non-toxic concentrations of Asiaticoside. Behavioural, biochemical, and cellular assays were performed to validate the developed model. Antioxidant potency was identified by testing the levels of lipid peroxidation, superoxide dismutase and xanthine oxidase in the brain homogenate. Iron chelating potency was assessed in Perl’s-stained brain tissue sections. The anti-inflammatory potency was observed by measuring fold change in the expression of inflammatory gene IL-1β in brain homogenates by RT-PCR method and by immunohistochemistry techniques. The neuroprotective role of the compound was studied by fluorescent staining of the brain whole cell suspension to check apoptotic and necrotic population post-treatment.

Results and Discussion

The iron accumulation in tissue sections and brain homogenates reduced 3-fold post treatment. With 600 μg/mL concentration of Asiaticoside, the malondialdehyde and xanthine levels reduced 3-fold, and scavenging activity was found to increase after 24 hr, indicating antioxidant potency. This concentration also resulted in one third reduction in the expression of pro-inflammatory cytokine IL-1β and reduced inflammatory cell infiltration showing anti-inflammatory properties. The compound was found to be neuroprotective by reducing the apoptotic cell population from 84% to 20% in comparison to untreated control. In addition, post-treatment the nitric oxide levels were restored better than the untreated control indicating modulation in cellular signalling mechanisms.

Conclusion

The adult Zebrafish ICH model was developed to present the multifaceted mammalian pathophysiology of ICH in one system. The phytocompound Asiaticoside is shown to modulate neuroinflammation through a cycle of biological properties thus being neuroprotective, opening avenues for further pre-clinical testing.