ABSTRACT
This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Cytokines/metabolism , Furans/pharmacology , Lignans/pharmacology , Lymphocyte Activation/drug effects , T-Lymphocytes/drug effects , Animals , Anti-Inflammatory Agents/isolation & purification , Antigens, CD/metabolism , Antigens, Differentiation, T-Lymphocyte/metabolism , Arctium/chemistry , Cell Cycle Checkpoints/drug effects , Cell Proliferation/drug effects , Female , Furans/isolation & purification , Interferon-gamma/metabolism , Interleukin-10/metabolism , Interleukin-2/metabolism , Interleukin-2 Receptor alpha Subunit/metabolism , Interleukin-4/metabolism , Interleukin-6/metabolism , Ionomycin/pharmacology , Lectins, C-Type/metabolism , Lignans/isolation & purification , Mice , Mice, Inbred BALB C , Plants, Medicinal/chemistry , T-Lymphocytes/cytology , T-Lymphocytes/immunology , Tetradecanoylphorbol Acetate/pharmacology , Tumor Necrosis Factor-alpha/metabolismABSTRACT
AIM: To investigate the effects of anhydroicaritin (AHI) on the immunologic function of mouse macrophages stimulated by lipopolysaccharide (LPS) in vitro and its related immunosuppressive mechanism. METHODS: Mouse bone marrow-derived macrophages were isolated. Then, the drug toxicology of different concentrations of AHI on macrophages was measured by CCK-8 assay. The amount of NO produced in macrophages was detected by Griess kit. The phagocytosis of macrophages to E.coli BioParticles was assayed by flow cytometry (FCM). The expression of CD69, which was the marker of early activation of macrophages, was measured by FCM combined with two-color immunofluorescent staining of cell surface antigen. Cytometric bead array (CBA) kit was used to detect the production of cytokines of macrophages stimulated by LPS. RESULTS: AHI (2.5, 5, 10 µmol/L) significantly reduced the production of NO in macrophages stimulated by LPS, and inhibited the phagocytosis of activated macrophages. The results of FCM analysis showed that AHI decreased proportions of CD69 on LPS-stimulated macrophages. Furthermore, AHI downregulated the secretion of cytokines of LPS-induced macrophages. CONCLUSION: AHI, which exhibits immunosuppressive effect on the mouse macrophages stimulated by LPS, is promising to be developed as an immunosuppressive reagent.
Subject(s)
Flavonoids/pharmacology , Flavonols/pharmacology , Macrophage Activation/drug effects , Macrophages/drug effects , Phagocytosis/drug effects , Animals , Antigens, CD/immunology , Antigens, CD/metabolism , Antigens, Differentiation, T-Lymphocyte/immunology , Antigens, Differentiation, T-Lymphocyte/metabolism , Cells, Cultured , Dose-Response Relationship, Drug , Escherichia coli/immunology , Flavonoids/chemistry , Flow Cytometry , Lectins, C-Type/immunology , Lectins, C-Type/metabolism , Lipopolysaccharides/pharmacology , Macrophage Activation/immunology , Macrophages/immunology , Macrophages/metabolism , Male , Mice , Mice, Inbred BALB C , Nitric Oxide/immunology , Nitric Oxide/metabolism , Phagocytosis/immunologyABSTRACT
The objective of this study is to illuminate the mechanism of biodegradation of triphenyltin (TPhT). The removal of TPhT by Klebsiella pneumoniae was, therefore, investigated through characteristics studies. The influences of the various parameters were also discussed. The results demonstrated that the cell, extracellular secretion and intracellular enzyme were the effective biomasses for the biodegradation of TPhT. At initial concentration of 3 mg x L(-1), 10.9%, 5.3% and 47.9% of TPhT could be degraded by these biomasses respectively at 30 degrees C within 2 hours under an rotary shaker at 120 r x min(-1). The experimental results also showed that the enzyme activity could be affected by the buffers, pH, temperature, metals and the concentration of TPhT. The degradation efficiency would reach the highest point at pH 8, and at the optimal temperature of 50 degrees C. Metals including Mg2+, Mn2+, Fe2+ and Fe3+ improved the enzyme activity at certain concentrations. In the presence of 15 mg x L(-1) of Mg2+, the removal percentage of TPhT was up to 73.8%. It suggested that the metals activated the enzyme and interacted with the TPhT enabling its removal during the biodegradation process. Linear plots of removal ratios versus concentrations of TPhT meant that the biodegradation fitted the Michaelis-Menten model. The Vmax and Km of this biodegradation were 0. 15 mg x (L x min)(-1) and 47.1 mg x L(-1), respectively.
