【摘要】 目的 研究白术内酯Ⅰ对白塞氏病小鼠脾脏辅助性T细胞(T helper cell, Th)17/调节性T细胞(T regulatory cell,Treg)平衡的影响。方法 将40只小鼠随机分为对照组,模型组,白术内酯I组,Jagged 1组,每组各10只,其中除对照组外,其余组均采用虾原肌球蛋白致敏建立白塞氏病模型。采用Western blot法检测小鼠脾脏中Notch1和Hes1的表达水平及Th17关键调控因子维甲酸相关孤儿受体(retinoic acid-related orphan receptor, ROR)γt和叉头框(forkhead box, Fox)p3的表达;采用流式细胞术检测小鼠脾脏中Th17和Treg的含量;采用ELISA法分别检测各组小鼠脾脏中Th17细胞因子白细胞介素(interleukin, IL)-17、IL-22的水平及Treg细胞因子IL-4和IL-10的水平;TUNEL法检测脾脏细胞凋亡;免疫组化法检测小鼠脾脏半胱氨酸天冬氨酸蛋白酶(caspase)-3的表达。结果 与对照组比较,模型组Th17细胞比例、Notch1、Hes1、IL-17、IL-22、RORγt和caspase-3水平以及TUNEL阳性细胞数升高,Treg细胞比例、IL-4、IL-10和Foxp3水平降低(P<0.05);与模型组比较,白术内酯Ⅰ组中Th17细胞比例、Notch1、Hes1、IL-17、IL-22、RORγt和caspase-3水平以及TUNEL阳性细胞数降低,Treg细胞比例、IL-4、IL-10和Foxp3水平升高(P<0.05);与白术内酯Ⅰ组比较,Jagged 1组Th17细胞比例、Notch1、Hes1、IL-17、IL-22、RORγt和caspase-3水平及TUNEL阳性细胞数升高,Treg细胞比例、IL-4、IL-10和Foxp3水平均降低(P<0.05)。结论 白术内酯Ⅰ能够改善原肌球蛋白诱导的白塞氏病小鼠脾脏Th17/Treg细胞比例失调,降低白塞氏病小鼠脾脏细胞的凋亡。更多还原

【Abstract】 Objective This study aims to investigate the effect of atractylenolide Ⅰ on the balance of helper T cells(Th)17/regulatory T cells(Treg) in the spleen of mice with Behcet’s disease. Methods 40 mice were randomly divided into the control group, model group, atractylenolide Ⅰ group, and Jagged 1 group, with 10 mice in each group. Except for the control group, the other groups were sensitized with shrimp tropomyosin to establish Behcet’s disease models. Western blot was used to detect the expressions of Notch1, Hes1, Retinoic acid-associated orphan receptor γt(RORγt), and forkhead box P3(Foxp3) in the spleen of mice. Flow cytometry was used to detect the content of Th17 and Treg in the spleen of mice. ELISA was used to detect the levels of interleukin(IL)-17, IL-22(Th17 cytokines) and IL-4, IL-10(Treg cytokines) in the spleen of mice in each group. Tunel was used to detect the apoptosis of spleen cells.Immunohistochemical method was used to detect the expression of caspase-3 in the spleen of mice. Results Compared with the control group, the cell proportion of Th17, the levels of Notch1, Hes1, IL-17, IL-22, RORγt, and caspase-3, the number of TUNEL-positive cells in the model group were increased(P<0. 05). While the cell proportion of Treg, the levels of IL-4, IL-10, and Foxp3 was decreased(P<0. 05). Compared with the model group, the cell proportion of Th17,the levels of Notch1, Hes1, IL-17, IL-22, RORγt, and caspase-3, the number of TUNEL-positive cells in the atractylenolide Ⅰ group were decreased(P<0. 05). While the cell proportion of Treg, the levels of IL-4, IL-10, and Foxp3 was increased(P<0. 05). Compared with the atractylenolide Ⅰ group, the cell proportion of Th17, the levels of Notch1, Hes1, IL-17, IL-22, RORγt, and caspase-3, the number of TUNEL-positive cells in the Jagged 1 group were increased(P<0. 05). While the cell proportion of Treg, the levels of IL-4, IL-10 and Foxp3 were decreased(P<0. 05).Conclusion Atractylenolide I can improve the tropomyosin-induced imbalance of cell proportion of Th17/Treg in the spleen of mice with Beh?et’s disease, and alleviate the apoptosis of spleen cells in mice with Behcet’s disease.更多还原