Cowanin induces apoptosis in breast cancer cells via Bcl-2 signaling pathway.

OBJECTIVES: This study aimed to investigate the effect of cowanin the mechanism of cowanin toward cell death and BCL-2 protein (antiapoptotic) expression of T47D breast cancer. METHODS: The cell death was evaluated by double staining, namely acridine orange and propidium iodide, and then observed under a fluorescence microscope. Meanwhile, the BCL-2 protein expression was determined by western blotting with measurement of protein area and protein density. RESULTS: The result found T47D breast cancer cells were viable, apoptosis, and necrosis after treatment with cowanin. The average viable cells, apoptosis, and necrosis percentages were 54.13 %, 45.43 %, and 0.44 %, respectively. Statistical analysis showed cowanin could significantly induce death in T47D breast cancer cells by apoptosis (p<0.05). It was also revealed that cowanin and positive control (doxorubicin) treatment had a significantly decreased protein area and protein density (p<0.05). CONCLUSIONS: It can be concluded that cowanin can induce death in T47D breast cancer cells by apoptosis and affect the expression of Bcl-2 protein in T47D breast cancer cells.

In vitro immunomodulatory activity study of Garcinia cowa Roxb. fraction.

OBJECTIVES: The objective of this study is to determine the activity of Garcinia cowa Roxb. n-hexane, ethyl acetate, and butanol fractions as an immunomodulator in vitro and obtain the fraction that has the potential as an immunomodulator. METHODS: Raw 264.7 macrophages were used to asses G. cowa Roxb. immunomodulatory activity. The MTT assay was chosen to measure cell viability to evaluate the cytotoxic effect on cells. ELISA method was used to measure the concentration of Interleukin-6 (IL-6) and Tumor Necrosis Factor Alpha (TNF-α) secreted by cells after being treated with G. cowa Roxb. fraction. The neutral red uptake assay determined the effect of Garcinia cowa Roxb. on the phagocytic activity. RESULTS: After Raw 264.7 macrophages were given the Hexan fraction (Hex) at concentrations of 12.5 and 25 µg/mL, there was a decrease in the concentration of IL-6, TNF-α, and the phagocytosis index of cells. Administration of the Ethyl Acetate fraction (EtOAc) at concentrations of 12.5 and 25 µg/mL on cells caused a decrease in IL-6 and TNF-α levels but did not affect the phagocytosis index. There was an increase in the level of TNF-α and the phagocytosis index after being given the Butanol fraction (BuOH) with concentrations of 12.5 and 25 µg/mL but there was a slight decrease in the level of IL-6. CONCLUSIONS: Both Hex and EtOAc fractions could suppress immune responses through decreasing IL-6, TNF-α, and slightly decreased phagocytic activity. BuOH fraction could stimulate immunomodulatory activities through enhanced TNF-α levels and phagocytic index, but less potent in enhancing IL-6 production. The BuOH fraction could be developed as an immunostimulant.

Blood pressure and blood sugar-lowering effects of purified gambir on diabetic hypertensive Wistar Kyoto rats.

OBJECTIVES: Antioxidants protect people from diabetes and its cardiovascular complication. Purified gambir (Uncaria gambir Roxb.) is a potential medicinal plant for treating this condition based on the antioxidant activity of its catechin compound. This study tries to reveal the potential activity of purified gambir as a blood pressure-lowering drug while lowering blood glucose in diabetic hypertensive rats induced by oral NaCl-Prednisone and Alloxan. METHODS: Rats were induced by oral NaCl 0.8% and Prednisone 5 mg/kg BW for 14 days to obtain hypertensive condition. Alloxan 125 mg/kg BW was given intra peritoneal injection on the 8th day to obtain diabetic hypertensive condition. The animal was divided into five groups, normal control group treated with vehicle, treatment groups were treated with purified gambir at dose of 2.5; 5 and 10 mg/kg BW respectively, while the positive control group were treated with a combination of captopril-glibenclamide at dose of 2.25 and 0.45 mg/kg BW. All animals were treated orally for 14 days. Fasting blood glucose and cardiovascular parameters (SBP, DBP, MAP, HR, BF and BV) were measured on days 1, 3, 7, and 14. NO level were measured on day 0 and day 14. Data were analyzed using two-way ANOVA followed by Duncan Multiple Range Test. RESULTS: The purified gambir has blood pressure and blood sugar-lowering activity (p<0.05). The NO levels of the treatment group also increased significantly (p<0.05). CONCLUSIONS: This study indicated that purified gambir could be an alternative medicine to manage blood glucose and blood pressure in the diabetic hypertensive model.

Cytotoxic Properties and Complete Nuclear Magnetic Resonance Assignment of Isolated Xanthones from the Root of Garcinia cowa Roxb.

OBJECTIVE: To isolate compounds from the roots of Garcinia cowa and to evaluated their cytotoxic activity against breast (MCF-7), prostate (DU-145), and lung (H-460) cell lines. MATERIALS AND METHODS: The ground air-dried root was sequentially macerated with hexane, dichloromethane (DCM), ethyl acetate (EtOAc), and methanol. The DCM soluble extract was fractionated by vacuum liquid chromatography, column chromatography, and radial chromatography over silica gel with hexane, EtOAc and methanol as eluent in progressively increasing polarity manner; to yield three compounds. Their structures were elucidated based on their spectroscopic data and their comparison with those of the literature. The cytotoxicity of isolated compounds was carried out against human cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay. The extract was added at various concentrations (0.1, 1, 10 and 100 mg/ml). The level of cytotoxicity was determined by calculating the level of IC50 that was based on the percentage of the cell death following the 24 h incubation with the extract. RESULTS: Phytochemical study on the roots of G. cowa yielded rubraxanthone (3), cowanine (4) and 1,5-dihydroxyxanthone (5). Compound 4 with an IC50 value of 4.1 ± 1.0 µM, 5.4 ± 2.3 µM and 11.3 ± 10.0 µM against MCF-7, H-460, and DU-145, respectively while compound 3 was found to be in active. CONCLUSION: The results indicate that G. cowa roots could be important sources of natural cytotoxic compounds. SUMMARY: Isolation of cytotoxic compounds from Garcinia cowaCowanine is the active constituent from the roots of Garcinia cowaComplete nuclear magnetic resonance assignment of isolated compoundsMS fragmentation of rubraxanthone.