Libidibia ferrea (jucá), a Traditional Anti-Inflammatory: A Study of Acute Toxicity in Adult and Embryos Zebrafish (Danio rerio)

Abstract

The plant species Libidibia ferrea (Mart. ex Tul.) LP Queiroz var. ferrea basionym of Caesalpinia ferrea (Mart. ex Tul.) is used in various regions of Brazil in folk medicine in the treatment of several health problems, especially in acute and chronic inflammatory processes. Most of the preparations employed are alcoholic. Therefore, this study aimed to evaluate the acute toxicity of the hydroethanolic extract of fruits of Libidibia ferrea (EHEFLf) in zebrafish, emphasizing the possible changes in the organic-cellular level of the gills, liver, kidneys, and intestine and on embryos. The result obtained by LC-M/MS from EHEFLf indicated a high concentration of possible polyhydroxylated substances. EHEFLf, at a dose of 2 g/kg orally, produced non-significant alterations of the analyzed organs. However, for embryos, the treatment with different concentrations demonstrated heart toxicity that was concentration-dependent. There is no evidence of a correlation of the observed effects with the phytochemical composition, and considering the species of animal used, it can be suggested that the oral use of L. ferrea hydroethanolic extract has an acceptable degree of safety for use as an oral medicinal product. and embryo results have shown significant affinity to the heart; however, it is perceived to be related to the concentrations used.

Keywords: Libidibia ferrea; embryos; histopathology; toxicity; zebrafish.

Figure 1

Base peak chromatogram of the hydroethanolic extract of fruits of Libidibia ferrea (EHEFLf) by Liquid Chromatography-High Resolution Tandem Mass Spectrometry (LC-HRMS/MS).

Figure 2

Histopathological alteration index (HAI) of D. rerio gills, liver, intestine, and kidneys, after 48 h of EHEFLf or propylene glycol oral administration at a dose of 2 g/kg.

Figure 3

Histological alterations of D. rerio gills after oral treatment with 2 g/kg EHEFLf (A) and propylene glycol 1% (B). Gill filament of an animal treated with EHEFLf, where secondary lamellae (LS), primary lamellae (LP), and central venous sinus (SVC) were observed (A); gill filament of a vehicle-treated animal, where dislocation or elevation of secondary lamellae (DELS), capillary derangement (DC), capillary dilation (DiC), and epithelial cell hyperplasia at the base of secondary lamellae (HCBLS) were observed (B).

Figure 4

Histological changes of the D. rerio liver after oral treatment with 2 g/kg of EHEFLf (A) and propylene glycol 1% (B). Liver of an animal treated with EHEFLf, where hepatocytes (H), venous sinusoids (SV), glycogen (G), cytoplasmic vacuolization (VC), and biliary stagnation (EsB) were observed (A); liver of animal treated with vehicle where nuclear vacuolization (VN), cytoplasmic vacuolization (VC), and nuclear atrophy (NA) were observed (B).

Figure 5

Histological changes of the D. rerio intestine after oral treatment with 2 g/kg of EHEFLf (A) and propylene glycol 1% (B). Intestine of an EHEFLf-treated animal where stromal lymphocytic infiltration (SLI), distal of the epithelial lining of the intestinal apex (DEL), leukocyte infiltration (ILI), and goblet cell hyperplasia (HGC) were observed (A); intestine of animal treated with vehicle-control, where stromal lymphocytic infiltration (ILS), lamina propria detachment (DLP), and leukocyte infiltration (ILI) were observed (B).

Figure 6

Histological changes of the D. rerio kidney after oral treatment with 2 g/kg of EHEFLf (A) and propylene glycol 1% (B). (A) Kidney of an EHEFLf-treated animal where renal tubules (RT), hematopoietic tissue (HT), glomerulus (G), Bowman capsule (CB), and tubular lumen enlargement (ALT) were observed; (B) kidney of animal treated with the vehicle-control where loss of the cellular contour (LCC), cytoplasmic tubular cell degeneration (CDTC), mild tubular hyaline degeneration (DHTL), and tubular obstruction (OT) were observed.

Figure 7

Teratogenic effects on D. rerio 96 hpf embryo. (A) Embryo treated at a concentration of 500 mg/L, where cardiac edema (Ce) and yolk edema (Ye) were observed; (B) embryo treated at a concentration of 250 mg/L, where scoliosis (Sc) was observed.