ABSTRACT
OBJECTIVE@#Although Angelica archangelica is a medicinal and aromatic plant with a long history of use for both medicinal and food purposes, there are no studies regarding the antineoplastic activity of its root. This study aimed to evaluate the cytotoxicity and antitumor effects of the crude extract of A. archangelica root (CEAA) on breast cancer.@*METHODS@#The cytotoxicity of CEAA against breast adenocarcinoma cells (4T1 and MCF-7) was evaluated by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Morphological and biochemical changes were detected by Hoechst 33342/propidium iodide (PI) and annexin V/PI staining. Cytosolic calcium mobilization was evaluated in cells staining with FURA-4NW. Immunoblotting was used to determine the effect of CEAA on anti- and pro-apoptotic proteins (Bcl-2 and Bax, respectively). The 4T1 cell-challenged mice were used for in vivo assay.@*RESULTS@#Using ultra-high-performance liquid chromatography-mass spectrometry analysis, angelicin, a constituent of the roots and leaves of A. archangelica, was found to be the major constituent of the CEAA evaluated in this study (73 µg/mL). The CEAA was cytotoxic for both breast cancer cell lines studied but not for human fibroblasts. Treatment of 4T1 cells with the CEAA increased Bax protein levels accompanied by decreased Bcl-2 expression, in the presence of cleaved caspase-3 and cytosolic calcium mobilization, suggesting mitochondrial involvement in breast cancer cell death induced by the CEAA in this cell line. No changes on the Bcl-2/Bax ratio were observed in CEAA-treated MCF7 cells. Gavage administration of the CEAA (500 mg/kg) to 4T1 cell-challenged mice significantly decreased tumor growth when compared with untreated animals.@*CONCLUSION@#Altogether, our data show the antitumor potential of the CEAA against breast cancer cells in vitro and in vivo. Further research is necessary to better elucidate the pharmacological application of the CEAA in breast cancer therapy.
ABSTRACT
O ácido hialurônico de baixo peso molecular (AH) é proposto como agente que atua nas três fases da reparação tecidual. O objetivo deste trabalho foi avaliar se o AH é capaz de induzir proliferação celular de fibroblastos dérmicos humanos. Esses efeitos foram avaliados empregando-se as técnicas de azul de tripano e de brometo de [3-(4,5-dimetiltiazol-2yl)-2,5-difenil tetrazolium] (MTT),seguida de análise do ciclo celular, produção de colágeno e modelo de wound healing (WH). Após tratamento com AH, foi verificado aumento significativo na proliferação das células e nas fases S-G2-M do ciclo celular. Quando submetido ao teste de WH, a exposição a 0,2% de AH promoveu redução significativa da área da lesão, com taxas de migração celular de 74%. Os resultados confirmam a capacidade do AH em aumentar a proliferação de fibroblastos, o que é relevante para a fase proliferativa da cicatrização.
Subject(s)
Humans , Wound Healing , In Vitro Techniques , Hyaluronic AcidABSTRACT
Cicatrization can be divided into three phases: inflammation, fibroblastic, maturation and remodeling [1]. The extracellular matrix may be replaced by a stronger and more elastic connective tissue. In a scar collagen is the major component of the mature connective tissue [2]. In homeopathic area, the greater is the investigated segment ultra dilutions [6]. However, little research has been done to explore the effect of dynamized drugs in in vitro cell culture [4]. Using the Zincum metallicum 6CH and Calendula officinalis 6CH applied independently in different concentrations in fibroblast cultures sought to determine the increase in proliferative activity using techniques such as IC50, MTT, flow cytometry and quantification of collagen. As expected from the literature, ie both homeopathic according to the literature are used for treatments that Require skin healing, both showed increased proliferative activity, having Calendula most cellular response, presenting as cell cycle stimulating checked via flow cytometry.