γ-氨基丁酸及其受体调控CD8<sup>+</sup>T细胞活化

樊文梅; 高钰; 孙玉洁; 马锡慧; 何秀云; 肖漓; 石炳毅

doi:10.3969/j.issn.1674-7445.2017.02.010

γ-氨基丁酸及其受体调控CD8⁺T细胞活化

doi: 10.3969/j.issn.1674-7445.2017.02.010

100091 北京，解放军第 309 医院全军器官移植研究所研究室

基金项目:

国家自然科学基金 81270509

详细信息

通讯作者:
肖漓, Email:xiaoli1972@163.com

石炳毅, Email:shibingyi@medmail.com.cn

中图分类号: R617, R392.4
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- 文章访问数: 223
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- PDF下载量: 7
- 被引次数: 0
出版历程
- 收稿日期: 2016-12-02
- 网络出版日期: 2021-01-19
- 刊出日期: 2017-03-15

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

Organ Transplant Institute of People's Liberation Army, the 309^th Hospital of People's Liberation Army, Beijing 100091, China

More Information

Corresponding author: Xiao Li, Email:xiaoli1972@163.com; Shi Bingyi, Email:shibingyi@medmail.com.cn

摘要

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。
- CD8⁺T细胞 /
- CD3/CD28 /
- γ-氨基丁酸 /
- 移植免疫 /
- GABA-A受体 /
- GABA-B受体 /
- 细胞增殖 /
- T细胞活化 /
- CD152
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.
- CD8⁺T cell /
- CD3/CD28 /
- γ-aminobutyric acid /
- Transplantation immunity /
- GABA-A receptor /
- GABA-B receptor /
- Cell proliferation /
- T cell activation /
- CD152

HTML全文

图 1 BrdU细胞增殖试剂盒检测GABA对CD8⁺T细胞增殖（S期）影响的流式分析图

A、C、E、G、I、K图分别为BrdU阴性对照、BrdU阳性对照、CD8+GABA（10 μmol/L）、CD8+CD3/CD28+IL-2、CD8+CD3/CD28+IL-2+GABA (10 μmol/L)、CD8+CD3/CD28+IL-2+GABA (50 μmol/L) 细胞增殖的结果；B、D、F、H、J、L图则为CD152⁺CD8⁺ T细胞的数量

Figure 1. Flow cytometric analysis of the effect of GABA on the cell proliferation of CD8⁺ T cells (S) detected by BrdU cell proliferation kit

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图 2 GABA对CD8⁺T细胞增殖（S期）和活化型CD8⁺T细胞影响的示意图

neg为阴性对照；CD8为BrdU阳性对照；CD8+G10为CD8+GABA（10 μmol/L）；CD8+CD3/28为CD8+CD3/CD28+IL-2；CD8+CD3/8/G10为CD8+CD3/CD28+IL-2+GABA (10 μmol/L)；CD8+CD3/28/G50为CD8+CD3/CD28+IL-2+GABA (50 μmol/L)；与CD8+CD3/28比较，^aP < 0.001

Figure 2. Digrams of effect of GABA on the cell proliferation (S) and activate state of CD8⁺T cells

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图 3 实时荧光定量PCR检测GABA受体的表达

α₃、α₅、β₂、β₃、γ₁、γ₂和θ均为GABA-A受体的不同亚型；B1a为GABA-B受体的1a亚型

Figure 3. Real-time PCR determination of the expression of GABA receptors

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