6.1.5. Toxicity Studies

Regarding the toxic effects of extracts obtained from *B. antiacantha*, a cytotoxicity test was carried out using *Artemia salina* nauplii, on which the alcoholic extracts from fruits and methanolic and alcoholic extracts from leaves of this plant showed toxicity, with LD50 values of 618.3 μg/mL, 275.9 μg/mL and 362.1 μg/mL, respectively [182]. In another study, hexane extracts in dichloromethane and ethyl acetate from *B. antiacantha* leaves showed toxicity against *Artemia salina*, with LD50 values of 53.9 μg/mL, 112.4 μg/mL and 241.6 μg/mL, respectively. For fruits, extract in dichloromethane was the only one with cytototoxic activity, with an LD50 of 29.8 μg/mL [193].

The hemolysis index promoted by the aqueous extract from *B. antiacantha* leaves and fruits was stipulated after analyzing lamb blood, observing total hemolysis in 0.85% and 1.00% dilutions for fruits, and 0.90% and 1.00% for leaves, while partial hemolysis occurred at 0.70% dilution for fruits and leaves. This hemolytic action was associated with the presence of saponin compounds in extracts, such as daucosterol (**63**), which were isolated from this species [196].

Saponins have cytotoxic activity, and this activity can occur through the promotion of autophagic cell death or cytoskeleton disintegration. In the case of hemolysis promoted by saponins, this may occur due to their ability to complex with cholesterol of the erythrocyte cell membrane, which results in the formation of pores in the membrane, increasing its permeability. In addition, some saponins, such as daucosterol (**63**) can act through signaling pathways such as PI3K/AKT, Wnt/β-catenin, promoting apoptosis. Autophagy induction can also occur by increasing the levels of light chain protein 3 (LC3) induced by some saponins. LC3 is associated with microtubules and is a marker for autophagy, being related to the formation of autophagic vacuoles [205,206].
