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Objective: To study the therapeutic effects of koumine on psoriasis in mouse models., Methods: The effects of koumine on epithelial cell mitosis and epidermal cell differentiation was evaluated by collecting the samples of the vaginal mucous and squamous epidermis at the tail of mice treated with methotrexate or koumine at different doses. The levels of interleukin (IL)-2 were detected with enzyme-linked immunosorbent assay., Results: High and intermediate doses of koumine showed remarkable inhibitory effect on mouse vaginal epithelial cell mitosis and promoted the formation of epidermal granular layer in the scales at the mouse tail. Three concentrations at 6, 30, 150 mg/kg of koumine decreased serum IL-2 level in the mice., Conclusion: The therapeutic effect of koumine against psoriasis is related to the inhibition of epidermal cell proliferation, promoting the formation of granular cells and decreasing the serum level of IL-2.

Objective: To assess the separation efficiency of magnetic-activated cell sorting in the purification of CD4+ T cells from murine spleen, and observe the effects of koumine on the proliferation of the separated cells., Methods: CD4+ T cells were isolated from murine spleen by magnetic-activated cell sorting (MiniMACS). Fluorescence-activated cell sortering was employed to determine the purity of CD4+ T cells before and after the separation procedure followed by evaluation of the cell viability using trypan blue staining. Concanavalin A- (ConA, 5 microg/ml) or phytahematoagglutinin (PHA,1 mg/ml)-induced murine T cells were treated with different concentrations of koumine (10-320 microg/ml), and their proliferation was determined by MTT colorimetry, and enzyme-linked immunosorbent assay was used to measure IL-2 level in the cell culture supernatant., Results: The purity of CD4+ T cells reached (90.3+/-5.8)% after the purification with a cell viability of (94.9+/-3.6)%. Koumine (20-320 microg/ml) dose-dependently inhibited ConA- or PHA-induced proliferation of murine lymphocytes as compared with the controls (P<0.05). Koumine (20, 100, and 200 microg/ml) significantly decreased the level of IL-2 in comparison with the control group (P<0.05)., Conclusions: CD4+ T cells of high purity can be obtained from murine spleen using MiniMACS without impairing the viability of the cells. Koumine significantly inhibits the proliferation of murine CD4+ T cells due to its immunosuppressive effect and inhibition of IL-2 secretion.

Objective: To identify the structure of the alkaloid extract of Gelsemium from east Guangdong province of China., Methods: Fourier transform infrared (FTIR) absorption spectrometry, hydrogen and carbon spectra of nuclear magnetic resonance (NMR) and distortionless enhancement by polarization transfer (DEPT) analysis were used for the structural identification of the alkaloid exstract of Gelsemium., Results: The frequency, intensity and shape of the extract's characteristic peaks in infrared absorption spectra (4,000.0-400.0 cm(-1)) and (1)H NMR, (13)C NMR were recognized and compared. The molecular structure of the sample was consistent with the theoretically derived model., Conclusion: The extract is structurally identical to koumine, which may provide evidence for its safe clinical application and establishment of Chinese medicine fingerprint database of Gelsemium.

Objective: To establish a method for isolating and identifying koumine., Methods: Koumine was first extracted from Gelsemium alkaloids by using chloroform, which was further separated and purified by column chromatograph, and identified by thin-layer chromatography., Results and Conclusion: Koumine crystals were isolated and purified from Gelsemium alkaloids effectively.

Objective: To develop a simple and efficient method for extracting Gelsemium alkaloids., Methods: Uniform design and orthogonal design were respectively utilized to optimize the 3 factors in the extraction process, namely the type of solvent used, the solid-to-liquid ratio in the extraction system and the reflux time., Results: Both of the design methods adopted trichloromethane as the solvent, with the solid-to-liquid ratio of 1:7.5 and reflux time of 3 h with 3 repetitions for the highest yield of alkaloids., Conclusion: Similar results are obtained from the two design methods for extracting Gelsemium alkaloids.