Efecto in vitro de los terpenos lupeol y casearina G sobre células sanguíneas y tumorales / In vitro effect of lupeol and casearin G on peripheral blood mononuclear and tumor cells

Background: The rainforest is an important source of natural compounds with therapeutic properties. Although there are many anti-inflammatory and antineoplastic drugs available to the clinician, there is an ongoing need for new therapeutic drugs with fewer serious adverse effects. Aim: To evaluate the in vitro cytotoxic effects of lupeol and casearin G on tumor cells, on phagocytic activity and nitric oxide (NO) production by blood mononuclear cells. Material and Methods: The cytotoxic effect of these compounds on cell lines MCF-7 (human breast adenocarcinoma) and PC-3 (human prostate cancer) was measured by a colorimetric assay (MTS/PMS) and the sulphorhodamine B assay. Peripheral blood mononuclear cells were obtained from eight healthy volunteers. The effect of these compounds on nitric oxide (NO) production was measured using the Griess reaction. Their effect on phagocytic activity of PBMC was also evaluated. Results: Lupeol (≥ 2 mM) resulted in a reduction of both the phagocytic index and the percentage of phagocytic monocytes and macrophages. Treatment of monocytes/macrophages with lupeol (72 µM) and casearin G (4 µM) reduced the production of NO. Neither lupeol (< 969 µM) nor casearin G (< 55 µM) had cytotoxic effects on PBMC. Casearin G showed both cytotoxic (IC50, LC50) and cytostatic (GI50) effects against tumor cells, PC-3 (IC50 = 12.5 µM; GI50 = 13.3 µM; LC50 = 51.9 µM) and MCF-7 (IC50 = 112.8 µM; GI50 = 11.8 µM; LC50 = 49.4 µM), as well as a hemolytic effect (≥ 182 µM). Conclusions: These observations indicate that lupeol and casearin G might be useful compounds in the preparation of anti-inflammatory drugs, whereas casearin G might be useful in the elaboration of antitumor drugs.

Lactonas sesquiterpénicas de las plantas Viguiera sylvatica y Decachaeta thieleana (Asteraceae) modulan la producción de óxido nítrico y la fagocitosis de macrófagos RAW

Sesquiterpene lactones of the plants Viguiera sylvatica and Decachaeta thieleana (Asteraceae) modulate nitric oxide production and phagocytosis of macrophages cell line RAW. Different species of the Asteraceae family are a potential source of sesquiterpene lactones with anti-inflammatory properties. Macrophages play a central role in the regulation of immune responses. In the present study, the in vitro effect of two sesquiterpene lactones, a millerenolide and a thieleanin, was assessed by measuring the production of nitric oxide (NO) by cell line RAW (murine macrophages) using the Griess reagent. Additionally, the effect of these sesquiterpene lactones on phagocytic capacity of latex particles and the reduction of nitroblue tetrazolium (NBT) were evaluated microscopically. Treatment of macrophages with >2.5 µg/ml of both sesquiterpene lactones, reduced the production of NO. A decreased number of macrophages able to reduce NBT were observed when these cells were treated with 3 µg/ml of millerenolide or 7.5 µg/ml of thieleanin. Treatment of macrophages with 4 µg/ml of millerenolide or 7.5 µg/ml of thieleanin, reduced the phagocytic capacity of macrophages. Cytotoxic effects on the macrophages were only observed when the concentration was increased to 8 µg/ml of millerenolide or 25 µg/ml of thieleanin. Our results suggest that these sesquiterpene lactones could be useful compounds in the elaboration of anti-inflammatory drugs. Rev. Biol. Trop. 56 (3): 1063-1073. Epub 2008 September 30.

Las plantas de la familia Asteraceae son una fuente potencial de lactonas sesquiterpénicas con propiedades antiinflamatorias. Los macrófagos son células que desempeñan una función central en la regulación de la respuesta inmune. Este estudio evaluó el efecto in vitro de dos lactonas sesquiterpénicas, un millerenólido y thieleanina, sobre la producción de óxido nítrico (NO) en una línea celular de macrófagos de ratón (RAW), utilizando el reactivo de Griess. Además, se estudió el efecto sobre la capacidad fagocítica de RAW, evaluando al microscopio la fagocitosis de partículas inertes de látex y la reducción de nitroazul de tetrazolio (NBT). Se observó que los macrófagos tratados con lactona sesquiterpénica (>2.5 µg/ml) disminuyeron la producción de NO. Además, se observó una disminución de la cantidad de macrófagos capaces de reducir NBT, después que fueron tratados con millerenólido (3 µg/ml) o thieleanina (7.5 µg/ ml). Por otro lado, la exposición a 4 µg/ml de millerenólido ó 7.5 µg/ml de thieleanina redujo la cantidad promedio de partículas de látex fagocitadas. No se observaron diferencias entre tratamientos y control en cuanto al porcentaje de células fagocíticas. Sólo se observaron efectos citotóxicos sobre los macrófagos, cuando la concentración de millerenólido se incrementó a 8 µg/ml o la de thieleanina se aumentó a 25 µg/ml. Estos resultados sugieren que el millerenólido y la thieleanina podrían ser compuestos útiles en la elaboración de drogas antiinflamatorias.

Actividad supresora del millerenólido sobre células mononucleares de sangre periférica humana / Millerenolide induces suppressive activity on human peripheral blood mononuclear cells

Background: Natural products are used in the production of therapeutic drugs due to their wide diversity and excellent adaptability to biological structures. Sesquiterpene ¡aciones are the active constituents of several plants from the Asteraceae family. Aim: To assess the in vitro effect of a sesquiterpene lactone (millerenolide). Material and methods: The drug effect was assessed measuring the proliferation of lymphocytes using the 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonylJ-2H-tetrazolium hydroxide (XTT) technique. Changes on the cell cycle were analyzed on a FACSort flow cytometer The effect of millerenolide on the production of nitric oxide (NO) by macrophages was evaluated using the Griess reagent. Additionally, phagocytosis of latex particles and nitroblue tetrazolium (NBT) reduction by macrophages were evaluated microscopically. Results: Treatment of human peripheral blood mononuclear cells (PBMC) with millerenolide decreases the proliferation of lymphocytes, decreases the percentage of cells in S, and G2/Mphases, and increases the proportion of cells in GO/Gl phase. Treatment of macrophages with millerenolide, reduces the production of NO, the phagocytic capacity and the number of cells able to reduce NBT. Cytotoxic effects of the lactone on human PBMC were only observed when the concentration was increased to 6 fig/ml. Conclusions: Millerenolide could be considered as a potential therapeutic agent with immunosuppressiveproperties.