Anthocyanins from black rice (Oryza sativa) promote immune responses in leukemia through enhancing phagocytosis of macrophages in vivo

Abstract

Rice is a staple food in numerous countries around the world. Anthocyanins found in black rice have been reported to reduce the risk of certain diseases, but the effects of crude extract of anthocyanins from Asia University-selected purple glutinous indica rice (AUPGA) on immune responses have not yet been demonstrated. The current study aimed to investigate whether AUPGA treatment could affect immune responses in murine leukemia cells in vivo. Murine acute myelomonocytic leukemia WEHI-3 cells were intraperitoneally injected into normal BALB/c mice to generate leukemia mice. A total of 50 mice were randomly divided into five groups (n=10 in each group) and were fed a diet supplemented with AUPGA at 0, 20, 50 or 100 mg/kg for three weeks. All mice were weighed and the blood, liver and spleen were collected for further experiments. The results indicated that AUPGA did not significantly affect animal body weight, but significantly increased spleen weight (P<0.05) and decreased liver weight (P<0.05) when compared with the control group. AUPGA significantly increased the T cell (CD3) population at treatments of 20 and 100 mg/kg (P<0.05). However, it only significantly increased the B cell (CD19) population at a treatment of 20 mg/kg (P<0.05). Furthermore, AUPGA at 50 and 100 mg/kg significantly increased the monocyte (CD11b) population and the level of macrophages (Mac-3; P<0.05 for both). AUPGA at 50 and 100 mg/kg significantly promoted macrophage phagocytosis in peripheral blood mononuclear cells (P<0.05), and all doses of AUPGA treatment significantly promoted macrophage phagocytotic activity in the peritoneum (P<0.05). AUPGA treatment significantly decreased natural killer cell activity from splenocytes (P<0.05). Finally, AUPGA treatment at 20 mg/kg treatment significantly promoted T cell proliferation (P<0.05), and treatment at 50 and 100 mg/kg significantly decreased B cell proliferation compared with the control group (P<0.05).

Laminarin Promotes Immune Responses and Normalizes Glutamic Oxaloacetic Transaminase and Glutamic Pyruvic Transaminase Levels in Leukemic Mice In Vivo

“…Institute (Hsinchu, Taiwan, ROC). Cells were cultured with IMDM supplemented with 10% FBS, 2 mM L-glutamine and antibiotics (100 units/ml penicillin, 100 μg/ml streptomycin) in 75 cm 2 tissue culture flasks and placed at 37˚C in a humidified atmosphere of 5% CO 2 (21).…”

“…Group II mice received normal diet as positive control; group III, IV and V mice received laminarin at 1, 2.5 and 5 mg/ml with ddH 2 O, respectively, by oral gavage every 2 days for 14 days (total of seven times). All animals were individually weighed during the oral treatment, and at the end of treatment, all mice were weighed and sacrificed as described previously (21).…”

“…Leukocytes were collected and stained with phycoerythrin (PE)-labeled anti-mouse CD3 and CD19, and fluorescein isothiocyanate (FITC)-labeled antimouse CD11b and Mac-3 antibodies (BD Biosciences Pharmingen Inc.) for 30 min. All samples were washed with phosphate-buffered saline (PBS) and cell markers (populations) were analyzed by flow cytometry as previously described (21,22).…”

“…Measurements of macrophage phagocytosis. All macrophages were isolated from peripheral blood mononuclear cells (PBMCs) and peritoneum as described previously (21,22). All macrophages were placed in plates containing 50 μl of target FITC-labelled Escherichia coli and were mixed and then were analyzed for phagocytosis by using flow cytometery following the PHAGOTEST ® kit manufacturer's instructions (ORPEGEN Pharma Gesellschaft für biotechnologische, Heidelberg, Germany).…”

Laminarin Promotes Immune Responses and Reduces Lactate Dehydrogenase but Increases Glutamic Pyruvic Transaminase in Normal Mice In Vivo

“…Results indicated that laminarin reduced NK cell cytotoxic activity at a target ratio of 50:1. Use of Yac-1 cells as a target cells for examining NK cell cytotoxic activity measurement is well documented (15).…”