Figure 6

A comparative effect of banaba leaf extract and policosanol (alone or in combination) on the hepatic histology of the streptozotocin-treated hyperlipidemic zebrafish (n = 3). (A) Hematoxylin and eosin (H&E) staining (400× magnified): blue and red arrows highlight the infiltration of neutrophils and lipid droplets, respectively [100 μm, scale bar]. The H&E area in the black box is 1000× magnified and images (B) represent this magnified view. (C) Oil red O (ORO) staining. (D) Immunohistochemical (IHC) staining for the detection of interleukin (IL)-6. (E) Red conversion of the IL-6-stained area (brown color) using the Image J software with a brown color threshold value of 20–120; the red conversion was performed to improve the intensity of the IL-6-stained area. (F) Percentage neutrophil counts. A semiquantitative evaluation of neutrophils (stained with a dark purple color) was conducted through a microscopic analysis of a predefined region (1.23 mm²) across three independent sections, with five distinct areas assessed per section. (G,H) illustrate the quantification of ORO and IL-6-stained areas, respectively. Abbreviations: HCD (high cholesterol diet), STZ (streptozotocin), BNB (banaba), and PCO (policosanol). The symbols *** underline the statistical significance at p < 0.001vs the HCD+STZ group. †, †† and ††† indicate statistical significance at p < 0.05, p < 0.01 and p < 0.001 vs. the BNB+PCO group; ns represents a non-significant (p > 0.05) difference.