Anti-inflammatory effect of rosmarinic acid isolated from Blechnum brasiliense in adult zebrafish brain

Neuroinflammation has been associated to neurodegenerative disease development, with evidence suggesting that high levels of proinflammatory cytokines promote neuronal dysfunction and death. Therefore, it is necessary to study new compounds that may be used as adjuvant treatments of neurodegenerative diseases by attenuating the inflammatory response in the central nervous system (CNS). The aim of this study was to utilize the lipopolysaccharide (LPS) induction model of neuroinflammation to evaluate the modulation of inflammation by rosmarinic acid (RA) isolated from Blechnum brasiliense in adult zebrafish. First, we investigated the toxicity and antioxidant properties of fractionated B. brasiliense extract (ethyl acetate fraction- EAF) and the isolated RA in zebrafish embryos. Next, we developed a model of neuroinflammation induction by intraperitoneal (i.p.) injection of LPS to observe the RA modulation of proinflammatory cytokines. The median lethal concentration (LC₅₀) calculated was 185.2 ± 1.24 μg/mL for the ethyl acetate fraction (EAF) and 296.0 ± 1.27 μM for RA. The EAF showed free radical inhibition ranging from 23.09% to 63.44% at concentrations of 10-250 μg/mL. The RA presented a concentration-dependent response ranging from 18.24% to 47.63% at 10-250 μM. Furthermore, the RA reduced LPS induction of TNF-α and IL-1β levels, with the greatest effect observed 6 h after LPS administration. Thus, the data suggested an anti-inflammatory effect of RA isolated from B. brasiliense and reinforced the utility of the new model of neuroinflammation to test the possible neuroprotective effects of novel drugs or compounds.

Keywords: Blechnum brasiliense; Neuroinflammation; Rosmarinic acid; Zebrafish.