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

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20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(P27kipl)、细胞周期蛋白E(cyclin E)、cyclin A、cyclin B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(25±10)%,对照组分别为(5±4)%、(11±6)%, 两组比较差异有显著性(P均<0.01);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<0.01).哮喘组T淋巴细胞内P27kipl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<0.05);哮喘组cyclin E、cyclin A、cyclin B的表达水平分别为(25±24)%、(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<0.01).结论 T淋巴细胞内CCRP异常表达与发作期哮喘患者T淋巴细胞过度活化、增殖有关,将CCRP作为调控靶点,可望成为治疗哮喘的新途径.

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

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.