支气管哮喘患者T淋巴细胞的细胞周期分布及周期调节蛋白表达的意义

The significance of cell cycle distribution and expression of cell cycle regulatory proteins in T lymphocytes in patients with acute asthma attack

目的 通过检测T淋巴细胞的细胞周期分布及其细胞周期调节蛋白 (CCRP)的表达 ,探讨发作期支气管哮喘 (简称哮喘 )患者T淋巴细胞过度活化、增殖的调控机制。方法 采用碘化丙啶DNA染色法应用流式细胞仪分析 30例发作期哮喘患者 (哮喘组 )及 2 0名正常人 (正常组 )外周血T淋巴细胞的细胞周期分布 ;采用间接免疫荧光法 ,应用流式细胞仪同步测定相应T淋巴细胞内CCRP、依赖激酶抑制蛋白 (P2 7kipl)、细胞周期蛋白E(cyclinE)、cyclinA、cyclinB的表达水平。比较哮喘患者和正常人上述指标的差异。结果 哮喘组S期、S +G2 /M期的T淋巴细胞百分率分别为 (18± 9) %和(2 5± 10 ) % ,对照组分别为 (5± 4 ) %、(11± 6 ) % ,两组比较差异有显著性 (P均 <0 0 1) ;哮喘组G0 /G1期的T淋巴细胞百分率为 (76± 10 ) % ,对照组为 (90± 6 ) % ,两组比较差异有显著性 (P <0 0 1)。哮喘组T淋巴细胞内P2 7kipl的表达水平为 (4 0± 2 4 ) % ,对照组为 (6 7± 4 8) % ,两组比较差异有显著性(P <0 0 5 ) ;哮喘组cyclinE、cyclinA、cyclinB的表达水平分别为 (2 5± 2 4 ) %、(9± 7) %、(6 4± 5 9) % ,对照组分别为 (6± 5 ) %、(4± 4 ) %、(3 4± 1 6 ) % ,两组比较差异均有显著性 (P均 <0 0 1)。结论 T淋巴细胞?

Objective To explore the molecular regulatory mechanisms of excessive T lymphocyte activation and proliferation in patients with asthma attack by studying the cell cycle distribution and the expression of cell cycle regulatory proteins(CCRP) in peripheral blood T lymphocytes. Methods DNA of T lymphocytes from 30 patients with asthma attack and 20 normal subjects was stained with propidium iodide, and the cell cycle distribution was analyzed by flow cytometry. By indirect immunofluorescence, the expressions of P27kipl, cyclin E, cyclin A, and cyclin B in T lymphocytes were measured by flow cytometry. The difference between patients with asthma attack and normal subjects was analyzed. Results(The percentage) of T lymphocytes in the S phase and S+G_2/M phase was (18±9)% and (25±10)% respectively in the asthma group, which were significantly higher than those in the control group [(5±4)%, (11±6)%,all P<0.01 respectively]. The percentage of T lymphocytes in the G_0/G_1 phase in the asthma group was (76±10)%, which was significantly lower than that in the control group [(90±6)%,(P<)0.01]. The expression of P27kipl in T lymphocytes of the asthma group (4.0±2.4)% was lower than that of the control group [(6.7±4.8)%,P<0.05]. The expression of cyclin E, cyclin A, and cyclin B in T lymphocytes of the asthma group [(25±24)%, (9±7)% and (6.4±5.9)%,respectively] were significantly higher than those of the control group [(6±5)%, (4±4)% and (3.4±1.6)% respectively],the differences being all statistically significant(all P<0.01). Conclusions The abnormal expression of CCRP in T lymphocytes was related to the excessive activation and proliferation of T lymphocytes in patients with asthma attack.A new approach to asthma therapy may be developed with CCRP as a regulatory target. [