Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, 81744-176, Iran.
Isfahan Neurosciences Research Center, Alzahra Hospital, Department of Neurology, Isfahan University of Medical Sciences, Isfahan, Iran.
Inflammation. 2019 Feb;42(1):54-63. doi: 10.1007/s10753-018-0872-x.
Multiple sclerosis (MS) is a central nervous system autoimmune disease characterized by demyelination. Autoreactive T cells mainly interferon gamma (IFN-γ) producing T helper cells (Th1) have an important role in MS pathogenesis. Silymarin is a unique blend produced from milk thistle (Silybum marianum) plant which its imunomodulatory role has been indicated in studies. In the present study, the effects of silymarin on isolated Th1 cells were investigated in newly diagnosed MS patients and those who received betaferon. PBMCs were separated from newly diagnosed and IFN-β-treated MS patients. The Th1 cell isolation from PBMCs was performed using a human Th1 cell isolation kit. Th1 cells were cultured in the presence of silymarin (50, 100, and 150 μM for 48, 72, and 120 h). Th1 cell proliferation and CD69 expression were assessed by flow cytometry. Also, IFN-γ production and T-bet gene expression were measured by ELISA and real-time PCR respectively. In vitro cultured Th1 cells showed that silymarin suppresses Th1 cell proliferation dose and time dependently in newly diagnosed and IFN-β-treated MS patients in comparison to DMSO control. Also, CD69 expression as an early activation marker was changed after Th1 cell treatment with different doses of silymarin at different times. T-bet gene expression was significantly decreased in Th1 cells isolated from newly diagnosed and IFN-β-treated RRMS patients after treatment with silymarin compared to DMSO control. Additionally, IFN-γ production by Th1 cells was decreased after treatment silymarin in newly diagnosed patients; however, in IFN-β treated after 48-h treatment with silymarin, IFN-γ concentration was decreased at concentrations of 100 and 150 μM, and after 120 h, a significant increase was observed in the IFN-γ level at a concentration of 100 μM in comparison with DMSO. Our findings here clearly show that silymarin is an effective regulator for Th1 response in vitro condition. It not only suppresses Th1 proliferating activity but also inhibits T-bet gene expression and IFN-γ production by these cells.
多发性硬化症（MS）是一种以脱髓鞘为特征的中枢神经系统自身免疫性疾病。主要产生干扰素γ（IFN-γ）的自身反应性 T 细胞（Th1）在 MS 的发病机制中起着重要作用。水飞蓟素是一种从奶蓟草（Silybum marianum）植物中提取的独特混合物，其免疫调节作用已在研究中得到证实。本研究探讨了水飞蓟素对新诊断的 MS 患者和接受β干扰素治疗的患者分离的 Th1 细胞的影响。从新诊断和 IFN-β治疗的 MS 患者中分离 PBMC。使用人 Th1 细胞分离试剂盒从 PBMC 中分离 Th1 细胞。将 Th1 细胞在水飞蓟素（50、100 和 150 μM 分别孵育 48、72 和 120 h）存在下培养。通过流式细胞术评估 Th1 细胞增殖和 CD69 表达。此外，通过 ELISA 和实时 PCR 分别测量 IFN-γ产生和 T-bet 基因表达。与 DMSO 对照组相比，体外培养的 Th1 细胞显示水飞蓟素在新诊断和 IFN-β治疗的 MS 患者中以剂量和时间依赖性方式抑制 Th1 细胞增殖。此外，在不同时间用不同剂量的水飞蓟素处理 Th1 细胞后，CD69 表达作为早期激活标志物发生变化。与 DMSO 对照组相比，用水飞蓟素处理后，新诊断和 IFN-β治疗的 RRMS 患者分离的 Th1 细胞中 T-bet 基因表达显著降低。此外，在用水飞蓟素治疗后，新诊断患者的 Th1 细胞 IFN-γ产生减少；然而，在用 IFN-β治疗 48 h 后，在 100 和 150 μM 浓度下水飞蓟素降低 IFN-γ浓度，而在 120 h 后，在 100 μM 浓度下 IFN-γ水平显著增加与 DMSO 相比。我们的研究结果清楚地表明，水飞蓟素是体外条件下 Th1 反应的有效调节剂。它不仅抑制 Th1 增殖活性，还抑制这些细胞的 T-bet 基因表达和 IFN-γ产生。
