| Objectives:Swainsonine, the main toxic component of the poisonous plants as Astragalus and Oxytropis, is the main toxin causing movement and reproductive disorders of poisoning symptoms. The purpose is to study the regulatory effect on Th1/Th2 cells balance and key cytokines reproductive immunity in serum of pregnancy Balb/c mice when swainsonine explosure so as to speculate the immune factors of the goat abortion. This study also lays a foundation for the mechanism of miscarriage.Methods:Balb/c of 8-week-old was used in this experiment. Three female and one male were keep in one cage for copulation.5 groups were divided to feed swainsonine solution of 0 mg/kg/d,0.2 mg/kg/d,0.4 mg/kg/d,0.8 mg/kg/d,1.6 mg/kg/d body weight, respectively,1/d, qd for 13d. At the twentieth day, histological section of ovarium and corpus luteum were made for judging vacuolation. Serum was get to measure the concentration of IL-1β, IL-4, IL-10, TNF-a and IFN-y by ELISA. The adjustment of SW to cytokines was analysised by expression of mRNA of IL-1β, IL-4, IL-10, TNF-a and IFN-y in spleen which were measured by RT-qPCR. Peripheral blood mononuclear cell were separated for flow cytometry after stimulation by PMA and Ionomycin. Using tricolor analytic method of flows cytometry to examinate Thl/Th2. And the effect of SW to Thl/Th2 was analysised.Results:Histopathological showed some cells in corpus luteum appear vacuolization in the dose of 0.2 mg/kg of swainsonine. With the increasing of dose, vacuolization was even more obvious and accompanied with cell nucleus swelling or karyopycnosis, and even cellular necrosis. This means that the pregnant mice showed the typical symptoms of locoism after feeding SW. From the date of the concentration of IL-1β, IL-4, IL-10, TNF-α and IFN-γ in the serum, we found that swainsonine could increase the concentration of IL-1β, TNF-α and IFN-γ significantly, and showed dose response relationship. There was a positive correlation between the dose of swainsonine and the concentration of IL-1β, TNF-α and IFN-γ. The serum concentration of IL-4 and IL-10 decreased after feeding SW. When the dose of Swainsonine was 0.2 mg/kg, the serum concentration of IL-4 was lowest. The concentration of IL-4 increased, when the dose increased. But it was still lower than the control group significantly. The concentration of IL-10 decreased with the dose increased. This showed negative correlation between the concentration of IL-10 and the dose of SW. Affter analysised the Th1/Th2 cell subgroup in the peripheral blood lymphocytes, we found that feeding the solution of swainsonine could increase the proportion of Th1 type cells of the mice and decrease the proportion of type Th2 cells. The balance of the Th1/Th2 subsets was broken down. With the dose of SW increased, the thl/th2 paradigm was lean towards to Thl to a higher degree.Conclusion:SW has immunomodulatory effects. It can promote the Thl cell proliferation and differentiation and increase the concentrations of IL-1β, TNF-α and IFN-γ in serum, and inhibite the proliferation and differentiation of the Th2 cells, reduce the levels of IL-4 and IL-10 cytokines. This suggest that SW lead the confusion of reproductive immunity of the pregnancy animal may be one of the reason that locoweed can cause abortion. |
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