γ-氨基丁酸及其受体调控CD8+T细胞活化

Activation of CD8+T cells regulated by γ-aminobutyric acid and its receptors

  • 摘要:
      目的  探讨γ-氨基丁酸(GABA)及受体对CD8+T细胞增殖能力的影响。
      方法  采用CD8+T细胞磁珠分选试剂盒分选Balb/c小鼠脾脏的CD8+T细胞;在CD3/CD28活化磁珠的作用下,以不同浓度的GABA刺激CD8+T细胞,通过5-溴脱氧尿苷(BrdU)标记和流式细胞术检测CD8+T细胞的增殖情况;通过实时荧光定量聚合酶链反应(PCR)比较CD8+T细胞活化前后GABA-A、GABA-B受体的表达。
      结果  流式检测结果显示,GABA可抑制已活化的CD8+T细胞的增殖,表现为CD152+CD8+T细胞明显减少;实时荧光定量PCR结果显示,仅在CD8+T细胞活化时表达的GABA-A受体亚型为α2、α6和GABA-B受体亚型为1a;CD8+T细胞活化后有明显增加的GABA-A受体亚型有α3、α5、β2、β3、γ1、γ2和θ,而GABA-B2R和GABA-B1b则在活化前和活化后均无表达。
      结论  GABA可抑制已活化的CD8+T细胞的增殖,其活化受GABA相关受体所调节,但其具体机制还有待于进一步探讨。

     

    Abstract:
      Objective  To evaluate the effect of γ-aminobutyric acid (GABA) and its receptors upon the proliferation of CD8+T cells.
      Methods  The splenic CD8+T cells of Balb/c mice were obtained by CD8+T cell magnetic bead sorting kit. Under the effect of CD3/CD28-activated magnetic bead, CD8+T cells were stimulated by GABA of different concentrations. 5-bromo-2-deoxyuridine (BrdU) labeling and flow cytometry were performed to detect the proliferation of CD8+T cells. The expression levels of GABA-A and GABA-B receptor before and after CD8+T cell activation were compared by fluorescent quantitative real-time polymerase chain reaction (PCR).
      Results  Flow cytometry result revealed that GABA could inhibit the proliferation of activated CD8+T cells, manifested as significant decrease in the quantity of CD152+CD8+T cells. Fluorescent quantitative real-time PCR demonstrated that GABA-A receptor subtypes α2, α6 and GABA-B receptor subtype 1a were expressed only when the CD8+T cells were activated. After CD8+T cell activation, the quantity of GABA-A receptor subtypes α3, α5, β2, β3, γ1, γ2 and θ were significantly increased, whereas the quantity of GABA-B2R and GABA-B1b did not significantly differ before and after CD8+T cell activation.
      Conclusions  GABA can suppress the proliferation of activated CD8+T cells. The activation of CD8+T cells is regulated by GABA receptors. However, the underlying mechanism remains to be elucidated.

     

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