Guazuma ulmifolia, or commonly known as the bastard cedar, has many pharmaceutical activities. Therefore, it is claimed as a source of various plant-based medicines. This research was purposed to identify the antioxidant activities of the ethanolic extract of G. ulmifolia (EEGU) by phytochemical screening assay, total flavonoid and total phenolic testing, and comparative analysis between the antioxidant activities of EEGU and epicatechin. The qualitative phytochemical screening assay of EEGU detected the availability of phenols, flavonoids, alkaloids, tannins, and terpenoids, but not saponins and triterpenoids. Meanwhile, the total phenolic content was 32.24 µg GAE/mg extract, and the total flavonoid content was 6.48 µg QE/mg extract. The role of antioxidants examined by FRAP, DPPH, H 2 O 2 , and ABTS assays. These assays are proved that the IC 50 values of EEGU are higher than epicatechin. For DPPH scavenging, H 2 O 2 scavenging, and ABTS reduction activities, EEGU resulted IC 50 45.70 μg/mL, 162.93 μg/mL, and 35.96 μg/mL, while epicatechin only yielded IC 50 0.56 μg/mL, 57.91 μg/mL, and 16.74 μg/mL respectively. Otherwise, the highest reduction in FRAP activities were shown at 50 μg/mL concentration of epicatechin and EEGU were 236.33 and 202.71 µM Fe (II)/µg respectively. Based on these results, EEGU is concluded as an active natural product because it exhibited antioxidant activities.
Introduction: Breast cancer is one of the leading cause of cancer deaths in women. Metastasis in BC is caused by immuno- surveillance deficiency, such NK cell maturation, low NK activity and decreasing cytotoxicity. This study was performed to improve activating receptors and cytotoxicity of NK cells using interleukins (ILs). Methods: Human recombinant IL-2, -15, and -18 were used to induce NK cells. We measured the activating and inhibiting receptors, proliferation activity of NK cells, and the cytotoxicity of NK cells on BC cells (MCF7). The effects of ILs were tested on the NK cell receptors CD314, CD158a and CD107a with flowcytometry, proliferation at various incubation times with 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and concen- trations of TNF-α and IFN-γ by NK cells with ELISA. Results: ILs increased NK cell receptor levels (CD314, CD158a, and CD107a) at 24 hours of incubation. ILs increased NK cell viability, which increased with longer incubation. Moreover, ILs-induced NK cells inhibited proliferation in MCF7 cells, as well as increased TNF-α, IFN-γ, PRF1 and GzmB secretion. Conclusion: IL-2, IL-15, and IL-18 improved activating receptors and proliferation of NK cells. IL-induced NK cells in- creased TNF-α, IFN-γ, PRF1 and GzmB secretion and cytotoxic activity on BC cells. High NK cell numbers increased BC cell growth inhibition. Keywords: Activator; breast cancer; interleukins; natural killer; receptor.
Aging is a natural process in human life and is triggered by the presence of free radicals (ROS). The use of antioxidants from natural ingredients is one of the breakthroughs to overcome aging and counteract the harmful effects caused by the free radicals. This study aimed to determine and compare the antioxidant activity of H2O2 scavenging and hyaluronidase inhibition of red dragon fruit peel extract (DFPE) and kaempferol-3-o-rutinoside (KOR) compounds. Dragon fruit peel extract (DFPE) is obtained through extraction by maceration method using 70% ethanol solvent. The design of this study included antioxidant and anti-aging activity assay of EKBN and KOR at the series concentration of 15.63; 31.25; 62.50; 125; 250; 500 µg/mL through H2O2 scavenging, as well as the DFPE and KOR hyaluronidase inhibition assay at the series concentration of 5.21; 10.42; 20.83; 41.7; 83.33; 166.67 µg/mL. EKBN shows that the average activity of H2O2 scavenging is lower than KOR. In addition, the IC50 values of KOR for H2O2 scavenging is lower (351.46±2.30ug/mL) than DFPE (409.64±23.17ug/mL). While, KOR also has higher values of inhibitory activity than of the DFPE. However, the IC50 value of KOR for hyaluronidase inhibition activity was 84.07±10.46µg/mL, equivalent to the IC50 value of DFPE (85.32±10.24µg/mL). The presence of antioxidant and anti-aging activity in the EKBN is probably caused by betalain and the KOR compound itself contained in red dragon fruit. The results of the paired-samples T-test on antioxidant activity and anti-aging of DFPE and KOR showed non-significant difference. Thus, DFPE has an equivalent antioxidant and anti-aging through H2O2 scavenging and hyaluronidase activity as possessed by the KOR compound.
Osteoarthritis (OA) is a chronic disease of the joints and bones due to trauma or other joint-related diseases (secondary). Synovial inflammation commonly causes disturbance in joint homeostasis, which is associated with OA. Enzymes such as aggrecanase and metalloproteinase generate cartilage damage, mediated by tumor necrosis factor (TNF-α) and interleukin (IL)-1. Proinflammatory cytokines, including TNF-α, IL-1β, and IL-6, are responsible for regulation of the extracellular matrix, cartilage degradation, and apoptosis of chondrocytes. This study aimed to observe the cell viability and expression level of matrix metalloproteinases (MMP-1 and MMP-3) and tissue inhibitor metalloproteinases (TIMP-1 and TIMP-2) in human chondrocyte cells (CHON-002) induced by IL-1β. CHON-002 was induced with IL-1β (0.1, 1 and 10 ng/mL) as an OA model. The viability of the cells was measured with a 3-(4,5dimethylthiazol-2-yl)-5-(3-carboxyme-thoxyphenyl)-2-(4-sulfophenyl)-2H-tetra zolium (MTS) assay, while expression of MMP-1, MMP-3, TIMP-1, and TIMP-2, was evaluated by RT-PCR. The viability of IL-1β-induced CHON-002 (CHON-002-IL-1β) cells at day 1 and 5 showed that treatment with up to 10 ng/mL of IL-1β was not toxic. Expression of TIMP-1 and TIMP-2 in CHON-002-IL-1β was lower compared to control, while that of MMP-1 and MMP-3 was higher compared to control. These results indicate that CHON-002 treated with 10 ng/mL IL-1β has expression patterns consistent with chondrocyte damage, so the CHON-002-IL-1β system may serve as a model for MMP induction in OA.
Breast cancer (BC) is the most prevalent type of cancer among women and one of the major causes of cancer mortality in women. Metastasis in breast cancer (BC) occurs due to immunosurveillance deficiency, including impairment of natural killer (NK) cell maturation. Conditioned medium (CM) from human Wharton's jelly mesenchymal stem cells (hWJMSC-CM) is known to possess anticancer activity. The CM of co-culture of human recombinant IL-2 treated NK cells and hWJMSCs is expected to boost anticancer activity toward BC cells which can be analyzed from the effect of CM towards secretion of effector molecules and expression of BC cell apoptosis-related genes, and cytotoxic granules in human recombinant IL-2 treated NK (IL-2 NK) and hWJMSCs (IL-2 hWJMSCs). TNF-α, IFN-γ, perforin, granzyme were measured by ELISA, while the inhibition of cell proliferation was measured by MTS assay and BC cell apoptosis by flow cytometry and apoptotic gene expression by RTPCR. CM from co-cultured hWJMSCs and IL-2 NK cells inhibited NK and BC cell proliferation, increased expression of Bax and p53 and decreased the number of Bcl-2 in BC cells. In conclusion, CM of co-culture IL-2 treated NK cells and hWJMSCs induce apoptosis in BC cells as indicated by increased Bax and p53 expression and decreased Bcl-2 expression.
Inflammation response is related with various diseases. One of the useful therapeutic method to suppress inflammatory mediator synthesis is by application of compounds isolated from herbal medicine as treatment for inflammatory diseases. The aim of this study was to analyse the anti-inflammatory activity of black soybean extract (BSE), daidzein, and genistein trough in vitro analysis of inflammatory mediators such as prostaglandin 2 (PGE2) and cytokines interleukin 1β (IL-1β), and tumor necrosis factor α (TNF-α). Safety of samples was determined by viability test using MTS (3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium). Concentration tested for viability assay were 40, 200, 1000 μg/mL for BSE, daidzein, and genistein. Anti-inflammation activity of samples was determined by ELISA quantification of PGE-2, TNF-α, and IL-1β in conditioned medium (CM) of supplemented pro-inflammatory activated RAW 264.7 cell. Inflammation on cells were induced by Lipopolysaccharide (LPS). BSE 1000 ug/ml, daidzein 1000 ug/ml, and genistein 1000 ug/ml treatments shows <80% cell viability average compared to control cell, indicating the treatments have cytotoxicity effect on RAW 264.7 cells. Hence, concentration used for treatments are 40 and 200 μg/mL for each sample. Genistein with concentration of 40 μg/ml treatment result shows highest anti-inflammatory activity which indicated from PGE-2, TNF-α, and IL-1β concentration. This study suggests that BSE, daidzein, and genistein with concentration of 40 and 200 μg/ml were safe to use for RAW 264.7 cell and genistein with concentration of 40 μg/ml have the best anti-inflammatory activity compared to daidzein and BSE.
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