Análise do perfil cromatográfico, do teor de marcador, e avaliação da atividade cicatrizante de feridas pépticas e de pele e da toxicidade do extrato aquoso das folhas de Bryophyllum pinnatum (Lam.) Pers. e Kalanchoe laciniata (L.) DC.: potencial aplicação do desenvolvimento de insumo

Abstract

Bryophyllum pinnatum and Kalanchoe laciniata (Crassulaceae), popularly known as “coirama” and “saião”, are used in folk medicine to treat gastric ulcers and in the skin wound healing. The present study aims to evaluate the chromatographic profile and the chemical marker content, in vivo anti-ulcer activities (oral route) and in vivo wound healing activities (topical route) of the aqueous extract of its leaves against a model of chronic gastric ulcer induced by acetic acid and an induction model of skin wounds, respectively, as well as to evaluate the acute and subchronic toxicity of B. pinnatum and K. laciniata leaf extracts (freeze dried extract) and the gastroprotective activity of the major flavonoid of B. pinnatum leaf extract, quercetin 3-O-α-L-arabinopyranosyl- (1→2)-O-α-L -rhamnopyranoside against acute gastric lesions induced by ethanol and indomethacin. Regarding the content of total phenols, B. pinnatum and K. laciniata extracts showed, respectively, levels of 80.87 ± 4.96 and 72.07 ± 4.78 mg (mg of gallic acid/g of dried extract) and total flavonoids of 38.97 ± 4.72 and 33 .83 ± 4.28 mg (mg of quercetin/g of dried extract). While the gel formulation contains B. pinnatum leaf extract at 5% had a total phenolic content of 2.77 ± 0.06 mg (mg of gallic acid/g of gel formulation) and total flavonoids of 1.58 ± 0,03 mg (mg of quercetin/g of gel formulation). Leaf extracts were analyzed by ultrafast liquid chromatography and the major flavonoids were identified. However, only the analytical method for B. pinnatum extract was validated according to national legislation and their major flavonoids quercetin 3-O-α-L-arabinopyranosyl-(1→2)-α-L-rhamnopyranoside (Bp1), kaempferol 3- O-α-L-arabinopyranosyl-(1→2)-α-L-rhamnopyranoside (Bp2) and quercetin-3-Orhamnopyranoside (Bp3) showed, respectively, a content of 33.12 ± 0.056, 3.98 ± 0.049 and 4.26 ± 0.022 mg/g of dried extract. Both extracts did not show signs of acute or subchronic toxicity and estimated lethal dose was defined as > 2,000 mg/kg, according to the OECD guide. B. pinnatum and K. laciniata extrats (250 and 500 mg/kg) significantly reduced the ulceration index (p < 0.0001 for doses) in chronic gastric ulcer in vivo model. This effect was accompanied by a rebalancing of gastric oxidative stress with increased levels of glutathione and expression of superoxide dismutase (SOD), in addition to reduced levels of malonaldehyde (MDA). Modulation of inflammatory markers was also observed with the reduction of myeloperoxidase (MPO) activity, of interleukin-1β (IL 1-β) and tumor necrosis factor-α (TNF-α) levels, increased interleukin-10 (IL 10), decreased expression of cyclooxygenase-2 and transcription factor NF-kB (p65). Additionally, treatment with K. laciniata extract stimulated healing of gastric tissue with increased expression of vascular endothelial growth factor (VEGF) and stimulated restoration of mucus content observed in the periodic acid-schiff staining technique. Pre-treatment with Bp1 (5 mg/kg) reduced gastric lesions induced by ethanol (p < 0.0001) and indomethacin (p < 0.001), increased GSH levels in gastric tissue and decreased MDA levels, reduced MPO activity reduced IL 1-β and TNF-α levels, increased IL 10 levels and presented a cytoprotective effect observed in histopathological analysis. The gel formulations contains the extracts stimulated the skin wound healing through a reduction in the wound area accompanied by a decrease in inflammatory infiltrate, a reduction in tissue levels of IL 1-β and TNF-α, stimulation of angiogenesis and an increase in the expression of VEGF These findings may support the development of a new inputs or herbal medicine from B. pinnatum and K. laciniata leaf extrat as gastroprotective or in ulcers treatment and for topical use to stimulate the healing of skin wounds.