红花黄色素对实验性自身免疫性脑脊髓炎小鼠脾细胞的作用

Effects of safflower yellow pigment on the spleen cells of experimental autoimmune encephalomyelitis mice

目的:初步探讨红花黄色素(SY)对实验性自身免疫性脑脊髓炎(EAE)小鼠脾细胞作用的有效性及可能机制。方法:雌性C57BL/6小鼠采用髓鞘少突胶质细胞糖蛋白35-55诱导,建立EAE模型,免疫后第9天制备脾细胞,和/无红花黄色素培养。CCK-8法和CFSE法检测脾细胞增殖情况,ELISA法检测上清液中肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β、干扰素γ(IFN-γ)、IL-17及IL-4水平,流式细胞术检测M1型和M2型巨噬细胞表型的变化,Griess法检测上清液NO含量。结果:SY浓度为26.24mg/L时均可提高脾细胞增殖率(P<0.01),抑制上清液中致炎因子TNF-α、IL-1β、IFN-γ、IL-17及升高抗炎因子IL-4蛋白的表达(P<0.01),降低脾细胞中致炎巨噬细胞比率并升高抗炎巨噬细胞的比率(P<0.01,P<0.05),可降低细胞上清液中NO含量(P<0.01)。结论:SY对EAE小鼠脾细胞作用的有效性与调整细胞因子含量及调节脾中致炎与抗炎巨噬细胞的比例有关,SY还可以降低NO含量。

Objective： To investigate the efficacy and possible mechanism of safflower yellow pigment （SY） on spleen cells in experimental autoimmune encephalomyelitis （EAE） mice. Methods： Female C57BL/6 mice were induced with myelin oligodendrocyte glycoprotein 35-55 to establish an EAE model. Splenocytes were prepared on day 9 after immunization, with/ without safflower yellow pigment was cultured. The proliferation of splenocytes was detected by CCK-8 method and CFSE method. The supernatant of tumor necrosis factor alpha （TNF-α）, interleukin-lβ （IL-1β）, IFN-γ, IL-17 and IL-4 were detected by ELISA. Flow cytometry was used to detect the changes of M1 and M2 macrophage phenotypes, and the Griess method was used to detect the NO content in the supernatant. Results： When the concentration of SY was 26.24mg/L, the proliferation rate of spleen cells could be increased （P〈0.01）, and the inflammatory factors TNF-α, IL-1β, IFN-γ IL-17 could be inhibited and the expression of anti- inflammatory factor IL-4 protein in the supernatant could be decreased （P〈0.01）, and it could reduce the ratio of proinflammatory macrophages in spleen cells and increase the ratio of anti-inflammatory macrophages （P〈0.01, P〈0.05）, which could reduce NO content in the cell supernatant （P〈0.01）. Conclusion： The effectiveness of SY on spleen cells of EAE mice is related to the regulation of cytokine levels and the regulation of the ratio of proinflammatory and anti-inflammatory macrophages in the spleen. In addition, SY can also reduce the NO content.