sq-23377 and sinomenine

The degranulation of mast cells and basophils is often initiated by a number of pathophysiological responses, especially in allergic and inflammatory conditions. Efficient techniques and methods for determining the level of such degranulation are highly demanded for laboratory and clinical studies. In this work, a rapid and sensitive assay based on the particle analysis of granules in RBL-2H3 cells, a cell line widely used as a convenient model system to study the degranulation of mast cells and basophils, was developed to detect cell degranulation using a Nanosight NS300 in light scatter mode and dynamic light scattering (DLS) on a Malvern Zetasizer Nano-ZS instrument. Using this method, drug-induced mast cell degranulation and systemic anaphylaxis were efficiently determined both in cell culture medium and blood samples from animals in the current study. This promising method is expected to be widely used for screening anti-allergic and anti-inflammatory drugs both in vitro and in vivo models, as well as for determining the level of mast cell degranulation of the patients in the clinic.

Topics: Animals; Basophil Degranulation Test; Basophils; Biosensing Techniques; Calcium; Cell Degranulation; Cell Line, Tumor; Dynamic Light Scattering; Equipment Design; Exocytosis; Female; Ionomycin; Mast Cells; Morphinans; p-Methoxy-N-methylphenethylamine; Predictive Value of Tests; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Time Factors

The effect on lymphocyte proliferation of sinomenine, a pure alkaloid extracted from the Chinese medical plant Sinomenium acutum was investigated in vitro using mouse spleen cells and human peripheral blood mononuclear cells. It could be demonstrated that sinomenine markedly inhibited [3H]thymidine incorporation in mouse spleen cells activated with concanavalin A (IC50 = 400 microM) or by two-way mixed lymphocyte culture (IC50 = 60 microM) and also in human peripheral blood mononuclear cells activated with phytohemagglutinin, 12-O-tetradecanoylphorbol-13-acetate plus ionomycin, or mixed lymphocyte culture (IC50 ranging from 34 to 129 microM). Time kinetic experiments revealed that sinomenine was effective only when added within the first 48 h after the onset of mixed lymphocyte culture, which lasted for 5 days. Inhibition of lymphocyte proliferation by sinomenine was reversible. Accordingly, the drug showed no direct cytotoxicity in our cellular systems and had no inhibitory effect on the proliferation of the cytokine-independent growth of the human leukaemic T-cell lymphoblast cell line Jurkat. It can be considered that these anti-proliferative effects are part of the anti-inflammatory and anti-arthritic mechanisms of sinomenine obvious in clinical trials.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Division; Cell Survival; Concanavalin A; Depression, Chemical; Female; Humans; Ionomycin; Leukocytes, Mononuclear; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Morphinans; Phytohemagglutinins; Spleen; Tetradecanoylphorbol Acetate