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为了研究全反式维甲酸(all-trans retinoic acid,ATRA)对Th细胞分化的影响和可能的机制,无菌分离BALB/c小鼠脾细胞后,用2.5μg/ml的ConA刺激脾细胞,在不同时间加入不同剂量的ATRA,72 h后收集细胞,用3H-TdR掺入法测淋巴细胞增殖活性,用RT-PCR方法检测在Th细胞分化中起作用的细胞因子的mRNA表达水平。结果ATRA呈剂量依赖抑制ConA诱导的淋巴细胞的活化,10~(-5)mol/L的ATRA对ConA诱导的淋巴细胞活化的抑制作用最强;此抑制作用与作用时间呈相关性,ConA活化淋巴细胞24 h后加入ATRA对淋巴细胞活化的抑制作用最显著,并且淋巴细胞的活化程度越低,ATRA对其的抑制作用越强。ATRA可增强经ConA活化的淋巴细胞Th2型细胞因子(IL-4、IL-6)mRNA的表达水平,而Th1型细胞因子(IFN-γ、TNF-α)mRNA的表达水平显著性降低(P<0.05)。上述结果表明ATRA总体上可抑制ConA刺激的淋巴细胞增殖活性,但却可增强,Th2细胞的分化。这一研究为Th细胞免疫偏离的基础与临床研究提供了依据。
In order to investigate the effect of all-trans retinoic acid (ATRA) on Th cell differentiation and possible mechanisms, splenocytes were stimulated with ConA at 2.5 μg / ml after aseptic isolation of splenocytes from BALB / c mice, Different doses of ATRA were added at different times, and the cells were harvested after 72 hours. The proliferation of lymphocytes was measured by 3H-TdR incorporation. The mRNA expression of cytokines that play a role in Th cell differentiation was detected by RT-PCR. Results ATRA inhibited the activation of lymphocytes induced by ConA in a dose-dependent manner. ATRA of 10 -5 mol / L had the strongest inhibitory effect on the activation of ConA-induced lymphocytes. The inhibitory effect was related to the action time and the activation of ConA The inhibition of lymphocyte activation by ATRA after 24 h was the most significant, and the lower the degree of activation of lymphocytes, the stronger the inhibition by ATRA. ATRA could enhance the expression of Th2 cytokines (IL-4, IL-6) mRNA by ConA-activated lymphocytes, while the expression of Th1 cytokines (IFN-γ, TNF- <0.05). The above results indicate that ATRA generally inhibits ConA-stimulated lymphocyte proliferation activity but enhances Th2 cell differentiation. This study provides a basis for the basic and clinical studies of Th cell immune deviation.