Centella asiatica extract ameliorates deoxygenation-induced neurological dysfunction in zebrafish larvae

Abstract

Background: Oxygen deprivation (OD) is a critical condition that can lead to brain damage and even death. Current hypoxia management approaches are limited in effectiveness. Centella asiatica (CA), known for its neuroprotective properties, offers a potential alternative for OD treatment.

Aims: This study aims to investigate the neuroprotective effects of CA on the expression of brain-derived neurotrophic factor (BDNF) and vesicular glutamate transporter 1 (VGLUT1) in zebrafish larvae under oxygen-deficient conditions.

Methods: Zebrafish embryos were subjected to low oxygen levels (1.5 mg/l) 0-2 hours post-fertilization (hpf) until 3 days post-fertilization (dpf), simulating the early stages of OD. Subsequent treatment involved varying concentrations of CA (1.25-5 µg/ml) up to 9 days post-fertilization. The expression levels of BDNF and VGLUT1 were measured using PCR methods. Statistical analysis was conducted using a two-way analysis of variance to evaluate the impact of CA on the expression of BDNF and VGLUT1 in zebrafish larvae aged 3 and 9 dpf in oxygen-deprived conditions.

Results: CA significantly influenced the expression of BDNF and VGLUT1 under OD (p < 0.001). An increase in BDNF expression (p < 0.001) and a decrease in VGLUT1 (p < 0.01) were observed in zebrafish larvae experiencing OD and treated with CA. There was no significant difference in BDNF and VGLUT1 expression across age variations in zebrafish larvae at 3 dpf and 9 dpf in the treatment groups (p > 0.05). CA concentration of 2.5 µg/ml effectively enhanced BDNF and reduced VGLUT1 in 3-9 dpf zebrafish larvae.

Conclusion: CA demonstrates potential as a neuroprotective agent, modulating increased BDNF expression and reduced VGLUT1 under OD conditions. These findings lay a foundation for further research in developing therapies for oxygen deficiency.

Keywords: Centella asiatica; Neuroplasticity; Neuroprotective; Neurotoxicity.

Fig. 1.. BDNF expression of embryonic zebrafish at 3, and 9 dpf in the treatment groups. Data are shown as the mean ± SD. The administration of CA with varying concentrations under OD conditions showed a significant difference in BDNF expression (p < 0.001). Still, there was no difference in BDNF expression based on the duration of administration (3 dpf and 9 dpf). There was no interaction between the treatment and the duration of administration. * (p < 0.05), dan ** (p < 0.01).

Fig. 2.. VGLUT1 expression of embryonic zebrafish at 3, and 9 dpf in the CA treatment groups under OD conditions. There was no significant difference in VGLUT1 gene expression with the timing of CA administration (ages 3 dpf and 9 dpf), and an interaction occurred between the treatment and the timing of CA administration (p > 0.05). A significant difference in VGLUT1 gene expression was caused by the concentration of CA treatment (p < 0.05). The administration of CA at a concentration of 2.5 μg/ml. **** (p < 0.0001) dan ns (not significance).