p38 MAPK对冲击波促进T淋巴细胞增殖和分泌IL-2的作用

doi:国家自然科学基金资助课题（30500132）

p38 MAPK对冲击波促进T淋巴细胞增殖和分泌IL-2的作用

赵毅¹，于铁成¹，金安¹，刘建国¹，郑学清²，徐莘香¹

1.吉林大学第一医院骨科，吉林长春130021；2.吉林大学第一医院碎石中心，吉林长春130021

收稿日期:2006-08-25 修回日期:1900-01-01 出版日期:2007-09-28 发布日期:2007-09-28
通讯作者: 于铁成

Effects of mitogen-activated protein kinase p38 on CD3/CD28-stimulated T-cell proliferation and IL-2 expression enhauced by shockwaves

ZHAO Yi¹, YU Tie-cheng¹, JIN An¹, LIU Jian-guo¹,ZHENG Xue-qing²,XU Xin-xiang¹

1. Department of Orthopedics, First Hospital, Jilin University, Changchun 130021,China；2. Lithotriptic Center, First Hospital, Jilin University, Changchun 130021,China

Received:2006-08-25 Revised:1900-01-01 Online:2007-09-28 Published:2007-09-28
Contact: YU Tie-cheng

摘要/Abstract

摘要： 目的：研究有丝分裂原激活蛋白激酶p38（p38 MAPK）对冲击波促进激活的T淋巴细胞增殖及分泌IL-2的作用。方法：预先用p38 MAPK抑制剂( SB203580，20 μmol•L^-1)分别与人外周血单个核细胞（PBMC）和Jurkat T细胞共同培养，同时设立不含SB203580的阴性对照组，再用冲击波和PHA或抗-CD3/抗-CD28抗体的亚刺激量作用，检测T淋巴细胞增殖和分泌IL-2的变化。采用免疫印迹法，用抗- p38MAPK抗体及抗-磷酸化的p38MAPK(Thr180/Tyr182)抗体，测定冲击波作用后Jurkat T细胞上的p38 MAPK的表达及磷酸化。结果：与未用冲击波作用组比较，被PHA激活的PBMC细胞，在能量密度为(0.180±0.015) mJ•mm^-2的冲击波作用100、150、200、250、300、330和360次时，细胞对³H-TdR掺入量明显增高（P<0.01）。加入SB203580的PHA激活的PBMC细胞，在上述同样强度的冲击波作用时，细胞对³H-TdR掺入量低于没有加入SB203580对照组（P<0.01）。与未受冲击波作用组比较，被CD3和CD28激活的Jurkat T细胞，在上述同样强度的冲击波作用时，细胞上清液中的IL-2活性明显增高（P<0.01）。加入SB203580的CD3和CD28激活的Jurkat T细胞，在该强度的冲击波作用时，细胞上清液中的IL-2水平低于比未加入SB203580的对照组（P<0.01）。50～250次的(0.180±0.015)mJ•mm²的低能冲击波可使Jurkat T细胞的p38MAPK磷酸化， p38MAPK的磷酸化程度随着冲击波作用次数的增加而增加。结论：SB203580可抑制低能冲击波对激活的T淋巴细胞的增殖及分泌IL-2作用；低能冲击波通过激活T淋巴细胞内的p38 MAPK，促进激活的T淋巴细胞增殖及分泌IL-2。

关键词: 有丝分裂原激活蛋白激酶p38, 冲击波

Abstract: Objective To investigate the effects of mitogen-activated protein kinase p38 (p38 MAPK) on CD3/CD28-stimulated T-cell proliferation and IL-2 expression which were enhanced by shockwaves. Methods Jurkat T cells or peripheral blood mononuclear cells (PBMC) were pretreated with the p38 MAPK-inhibitor (SB203580)(The Jurkat T cells or PBMC of the control groups were not pretreated with SB203580), then treated with shockwaves, and stimulated respectively with a suboptimal dose of anti-CD3 and anti-CD28 antibodies or PHA. Finally the IL-2 productions of Jurkat T cells or the proliferation of PBMC were respectively measured. The protein tyrosine phosphorylation of p38 MAPKin Jurkat T cells treated either with shockwaves or not was measured by Western blotting with anti-phosphotyrine antibodies(Thr180/Tyr182). The expressoins of p38 MAPK in Jurkat T cells treated either with shockwaves or not were determined by Western blotting with anti-p38 MAPK antibodies. Results Compared with negative control group without shockwave treatment, the ³H-TdR incorporation of the phytohemagglutinin-stimulated PBMC, which were treated with low dose shockwaves (LDSWs) of 100, 150, 200, 250, 300, 330 impulses at (0.180±0.015) mJ•mm², increased significantly (P<0.01). Compared with negative control group, the additions of SB203580 attenuated the IL-2 expression in the cellular supernatant of CD3/CD28-stimulated Jurkat T cells or the ³H-TdR incorporation of the phytohemagglutinin-stimulated PBMC, which were affected with LDSWs of 100, 150, 200, 250, 300, 330 impulses at (0.18±0.015) mJ•mm² (P<0.01). The activation of p38 MAPK under the condition without LDSWs was obviously lower than that with LDSWs of 100, 150, 200, and 250 impulses (P<0.01), and the activation of p38 MAPK increased with the shock-wave impulses.Conclusion SB203580 prevents the enhancing effect of LDSWs on CD3/CD28-stimulated IL-2 expression. In T cells, the shockwaves enhance IL-2 expression through activating p38 MAPK.

Key words: T cell proliferation, interleukin 2, mitogen-activated protein kinase activating p38

中图分类号:

R392.12

赵毅，于铁成，金安，刘建国，郑学清，徐莘香. p38 MAPK对冲击波促进T淋巴细胞增殖和分泌IL-2的作用[J]. J4, 2007, 33(5): 815-819.

ZHAO Yi, YU Tie-cheng, JIN An, LIU Jian-guo,ZHENG Xue-qing,XU Xin-xiang. Effects of mitogen-activated protein kinase p38 on CD3/CD28-stimulated T-cell proliferation and IL-2 expression enhauced by shockwaves[J]. J4, 2007, 33(5): 815-819.

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