ABSTRACT
The aim of this study was to investigate whether (-)-epicatechin (EC) can induce DNA damage and apoptosis of cancer cells in the spleen of rat with acute myeloid leukemia. Healthy and leukemic rats were given EC by gavage at a dose of 40 mg/kg b.w. for 22 consecutive days. Spleen cells were subjected for analysis of DNA damage and apoptosis. The amount of DNA damage was estimated by the comet assay, while apoptosis was examined by flow cytometry using Annexin V staining. Leukemic cells were identified in the spleen cells by indirect immunofluorescence using RM-124 antibody followed by flow cytometry analysis. The results show that EC did not affect DNA damage in the splenocytes of healthy rats, but significantly increased the extent of DNA strand breaks in the spleen cells of leukemic animals. EC administration to leukemic rats induced a significant increase in the level of Annexin V-positive leukemic cells, but the level of non-leukemic Annexin V-positive cells remained unchanged in comparison to control. The percentage of leukemic cells decreased significantly under EC influence comparing to the untreated group. The results of the study reveal that EC could be used as an effective supplement of standard therapy against acute myeloid leukemia.
Subject(s)
Antineoplastic Agents, Phytogenic/therapeutic use , Catechin/therapeutic use , Leukemia, Myeloid, Acute/drug therapy , Spleen/pathology , Animals , Annexin A5/chemistry , Apoptosis/drug effects , Body Weight/drug effects , Cell Line , Comet Assay , DNA Damage , Flow Cytometry , Fluorescent Antibody Technique, Indirect , Leukemia, Myeloid, Acute/pathology , Methylation , Organ Size/drug effects , Oxidation-Reduction , Phosphatidylserines/metabolism , Purines/chemistry , Rats , Spleen/drug effectsABSTRACT
Quercetin may have the opposite effect, namely anti- as well as pro-oxidant. The aim of this study was to assess the results of quercetin anti- and/or pro-oxidant activity in the bone marrow and spleen cells of rats. The experimental rats were treated daily, with quercetin in a dose of 8 or 80mg/kg b.w. by gavage for 40 days. The intracellular redox state in cells were assessed by measuring the ferric ion reducing antioxidant power (FRAP) level and malonodialdehyde concentration. HO-1 mRNA expression was examined with real-time PCR. The extent of DNA damage was determined by the alkaline-labile comet assay. A potential pro-apoptotic quercetin action was determined using the FITC-Annexin V kit. The quercetin and isorhamnetin concentrations in serum were analyzed by HPLC-ECD. MDA concentration and FRAP values, were significantly decreased in the spleen and bone marrow cells of rats treated with quercetin, in a dose of 80mg/kg b.w. in comparison with the control rats; no significant changes were observed after quercetin was administered in a dose ten times as low. Treatment with quercetin dose-dependently upregulated the expression of HO-1 mRNA in the bone marrow cells. Quercetin administration to the rats did not induce either DNA damage or apoptosis in the examined cells. The results of our study prove that changes in the antioxidant state, caused by quercetin, do not lead to DNA damage or exert any pro-apoptotic activity in vivo.
Subject(s)
Antioxidants/metabolism , Apoptosis/drug effects , Bone Marrow Cells/metabolism , DNA Damage , Quercetin/toxicity , Spleen/metabolism , Animals , Annexin A5 , Bone Marrow Cells/drug effects , Coloring Agents , Comet Assay , Flavonols/analysis , Flavonols/toxicity , Fluorescein-5-isothiocyanate/analogs & derivatives , Heme Oxygenase-1/metabolism , Lipid Peroxidation/drug effects , Male , Malondialdehyde/analysis , Propidium , Quercetin/analysis , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Rats , Rats, Inbred BN , Reverse Transcriptase Polymerase Chain Reaction , Spleen/cytology , Spleen/drug effectsABSTRACT
The aim of the experiment was to histochemically examine the activity of the following selected steroidogenic enzymes: delta5 3betaHSD and 17betaHSD and the housekeeping enzyme G6PDH, in experimental rats after subcutaneous injections of naringenin flavonoid, at a daily dose of 15 mg/kg of body mass. The enzyme activities were measured by the microdensitometric method. Additionally, radioimmunological assay for testosterone level was conducted in homogenates of testes. A significant decrease in the activity of 17betaHSD and of G6PDH was found, while the activity of delta5 3betaHSD was not significantly changed. The results permit a statement that naringenin causes minor changes in metabolic processes in the testes of rats but it does not significantly affect the synthesis of androgens.
