摘要
目的：研究有丝分裂原激活蛋白激酶p38（p38MAPK）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p38MAPK抑制剂（SB203580,20μmol·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p38MAPK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·mm^2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p38MAPK,促进激活的T淋巴细胞增殖及分泌IL-2。
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 a H-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 · mm^2, 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 · mm^2 （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.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2007年第5期815-819,共5页
Journal of Jilin University：Medicine Edition
基金
国家自然科学基金资助课题(30500132)
关键词
T淋巴细胞
有丝分裂原激活蛋白激酶p38
冲击波
shockwaves
T cell proliferation
interleukin 2
mitogen-activated protein kinase activating p38
