Oil-resin of Copaifera multijuga Hayne is popularly used due to its anti-inflammatory properties. In this study, we evaluate the effects of the bark ethanolic extract on Ehrlich tumor cells. For extract preparation, bark was removed from stem and chopped into smaller pieces. Most were dried and grounded. The first extract was made with hexane, followed by ethyl acetate. Absolute ethanol was added to the residue and extract was filtered and concentrated (EtOH). In vitro cytotoxic activity of the EtOH was analyzed on both spleen and tumor cells. For evaluation of in vivo activity, Ehrlich carcinoma bearing Swiss mice were daily treated with different concentration of EtOH during 30 days. Evaluation of in vitro cell viability showed that 1mg/ml extract is cytotoxic to spleen and tumor cells. Evaluation of in vivo tumor development showed that treatment with 200mg/kg of EtOH was able to reduce in 45.28% the tumor growth. Treatment for 7 days increased IL-12p70, IFN-γ and TNF-α production and for 14 days reduced the lymphoproliferative activity in both tumor bearing and mice healthy. Our results show that the ethanolic extract of copaiba bark affect Ehrlich tumor cells, reducing their cell viability in vitro as well as their development in vivo.
Copaifera multijuga, commonly known as copaiba, is popularly used in the form of tea for various conditions due to the presence of antioxidant substances in its composition, which protect cells against damage caused by free radicals. Its oleoresin is also used as an anti-inflammatory and antitumoral agent. The present study investigated the antioxidant effect of the ethanolic extract of copaiba stem bark on Swiss mice inoculated with solid Ehrlich tumors. Mice were inoculated subcutaneously with 1x106 Ehrlich’s tumor cells and treated via gavage with ethanolic extract of copaiba for thirty days, with doses varying between 100 and 200 mg kg-1. Biochemical analyses of enzymatic antioxidants [superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST)], non-enzymatic antioxidants [reduced glutathione (GSH) and ascorbic acid (ASA)], substances reactive to thiobarbituric acid (TBARS) and protein carbonylation (carbonyl) in different tissues were significantly affected. The extract administered at 200 mg kg-1 presented higher antioxidant capacity in the liver, increased CAT, GST, GSH and decreased TBARS, as well as increased CAT activity and protein carbonylation in brain tissue. The results showed that the copaiba extract was able to reverse the oxidative stress caused by solid Ehrlich tumor, probably due to the presence of antioxidant compounds, and had potential antineoplasic effect after a 30-day treatment.
The genus Duguetia, of the Annonaceae family, includes around 100 species that have been used in folk medicine to treat several diseases. Its phytochemical compounds have been researched for their antifungal, antioxidant, antigenotoxic, anti-inflammatory and cytotoxic properties. Therefore, the objective of the present study was to conduct a preliminary study on the biological effects of Duguetia sp extracts by evaluating their in vitro cytoto xic effect on different cell types. To prepare the methanolic extract (ME), the leaves were macerated with 80% methanol. Part of the ME was dissolved in 10% phosphoric acid, and the acidic aqueous solution was partitioned with dichloromethane. The organic phase was evaporated to obtain an acetogenin-rich extract (ACE). The extracts were diluted in RPMI culture medium , supplemented with 20% fetal bovine serum , and added to Erhlich tumor cells or mice spleen cells at different concentrations: the ACE e xtract a t 1.35 mg mL-1 , 0.67 mg mL-1 or 0,34 mg mL-1 , and the ME e xtract at 1.40 mg mL-1 , 0.70 mg mL-1 or 0,35 mg mL-1. After 24 h, cytotoxicity analysis was performed using the Trypan Blue exclusion method. The results demonstrated that the extracts are cytotoxic at all concentrations to Ehrlich tumor cells and mice spleen cells. According to the results, we concluded that the extracts of Duguetia sp had a cytotoxic effect on the studied cells in vitro. This was the first study to report the biological effect of this plant genus in this type of cells; however, further study is needed to determine the current species used and the compounds present in the extracts.
Euphorbia tirucalli L. is commonly used to treat various pathologies, including cancer. The objective of this study was to evaluate the effect of a crude extract of E. tirucalli on the development of Ehrlich solid tumors and immune function. We prepared an extract by macerating the aerial parts of the plant with 80% ethanol (1:10 g/v) for 15 days. Its effects on tumor development were assessed in male Swiss mice (n=10). The mice were injected subcutaneously with 10⁶ tumor cells and then treated by gavage daily with the extract (33.33, 67, and 133.34 mg kg-1) or saline (0.5% ethanol) for 30 days. The treatment had no toxic effect and did not reduce tumor growth. However, the weight of tumor mass was lowest in the mice treated with extract at 67 mg kg-1. Immunomodulatory activity was evaluated in mice (n=8), with and without tumors, treated with the extract (67 mg kg-1), under the same conditions as described above for 7 and 14 days. Lymphoproliferation, NK activity and IL-12, TNF-a, IFN-y, and IL-10 levels did not differ significantly between the groups. However, IL-4 levels were reduced in the E. tirucalli treated groups at 7 and 14 days when compared to the controls. We concluded that the extract is not toxic and cannot inhibit Ehrlich solid tumor development. Its immunomodulatory activity involves its ability to modulate IL-4 levels.
AIMS: Copaifera multijuga Hayne oleoresin is commonly used in traditional medicine owing to its anti-inflammatory, antiseptic, antitumor, and antibacterial properties. However, little is known about the effect of the compounds from the bark of this plant. In this study, the immunomodulatory effect of the ethanolic extract of C. multijuga bark via natural killer activity of non-adherent spleen cells of Ehrlich tumor-bearing mice was evaluated.METHODS: Male Swiss mice were inoculated subcutaneously with 1×106 Ehrlich tumor cells (Ehrlich and Ehrlich/C. multijuga group) or phosphate buffered saline solution (control group and C. multijuga group) and treated orally daily with C. multijuga extract (200 mg kg-1, 0.1 mL per mouse, for the Ehrlich/C. multijuga and C. multijuga groups) or phosphate buffered saline solution (control group and Ehrlich group). The four experimental groups consisted in eight mice each and were organized in two sets, one treated for seven days and another treated for 14 days, totalizing 64 mice throughout the experiment. Twenty-four hours after the last oral administration, the mice were euthanized and the spleen tissue was isolated to prepare a non-adherent spleen cell suspension and to evaluate natural killer activity. Data are presented as the cell lysis percentage of Yac.1 target cells by non-adherent spleen cells.RESULTS: Treatment for seven days increased natural killer activity in the Ehrlich/C. multijuga group (21.20±8.89, p<0.05) compared to the control group (3.14±2.71, p<0.05); however, this effect was not maintained in the groups treated for 14 days (Control: 6.02±6.98, Ehrlich: 4.82±7.72, C. multijuga: 2.07±2.10, Ehrlich/C. multijuga: 2.01±1.63, p>0.05).CONCLUSIONS: Treatment for seven days with an ethanolic extract of C. multijuga bark enhanced the natural killer activity of non-adherent spleen cells from Ehrlich tumor-bearing mice.
Ethnobotanical surveys have revealed the use of jatobá for the treatment of several diseases. This study determined the effect of plant extracts on the development of Ehrlich carcinoma. Male Swiss mice (n=6) were subcutaneously inoculated with 106 tumor cells and intragastrically administered ethanol (2 mg·mL-1, 5 mg·mL-1, or 10 mg·mL-1) or aqueous extracts of jatobá seed or bark for 90 days. Tumor development did not significantly differ between the groups studied; however, animals treated with the aqueous extract of the seed (2.205 mg·mL-1) had a reduction in tumor size compared to those treated with the aqueous extract of the bark (1.7 mg·mL-1). The treatment was not found to influence the survival of the animals studied. A new group of animals (n=7), with or without the tumor, received the aqueous extract of jatobá seed for 7, 14, and 30 days to evaluate oxidative stress. The extract reduced the thiobarbituric acid reactive substances levels at 7 days in the liver and kidneys, and 14 days in brain and renal tissue. Protein carbonylation levels were also reduced at 7 days in the liver and brain tissue and 14 days in the liver. The reduced glutathione levels diminished in animals treated for 7 and 14 days. We conclude that treatment with the aqueous extract of the jatobá seed presents promising activity in the reduction of oxidative stress.
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