论文部分内容阅读
目的探讨哮喘缓解期患者外周血 T 淋巴细胞亚群增殖规律及其分子机制。方法收集45例哮喘急性发作期患者(哮喘急发组)、哮喘缓解期患者(哮喘缓解组)和正常对照(对照组)的外周血标本(各15例),体外分离获得并培养3组患者的外周血 CD~+_4和 CD~+_8 T 淋巴细胞,用流式细胞术检测细胞周期时相分布、细胞周期调节蛋白[P27~(kipl)、细胞周期素(Cyclin)D、E]、PI_3K 和 STAT_6的表达水平。结果 (1)3组 CD~+_4 T 淋巴细胞的 G_0/G_1期分布比例、S 期分布比例,G_2/M 期和(S+G_2)/M 期比例差异有统计学意义(Z 值分别为12.35、8.05、9.16、12.80,均 P<0.05),3组 CD~+_8 T 淋巴细胞的 G_0/G_1期分布比例、S 期分布比例和(S+G_2)/M 期比例差异有统计学意义(Z 值分别为21.60、25.22和21.52,均 P<0.01);(2)3组 CD~+_4 T 淋巴细胞的 P27~(kipl)、Cyclin D 和 Cyclin E 表达水平差异有统计学意义(Z 值分别为10.63、10.66和6.64,均 P<0.05),3组 CD~+_8 T 淋巴细胞的 P27~(kipl)表达水平差异有统计学意义(Z 值为9.30,P<0.05);(3)3组 CD~+_4 T 淋巴细胞的 PI_3K、STAT_6蛋白表达水平差异有统计学意义(Z 值分别为9.04和18.08,P 值分别<0.05和0.01),哮喘缓解期患者处于增殖状态的 CD~+_4 T 细胞较正常对照者明显增多,与哮喘发作期患者相比处于增殖状态的 CD~+_4、CD~+_8 T 细胞显著减少。PI_3K 信号转导途径可能通过上调 CyclinD 和/或 CyclinE 影响 CD~+_4、CD~+_8 T 细胞增殖活化,而 JAK1-STAT_6信号转导途径可能通过下调 P27~(kipl)发挥作用。结论哮喘缓解期患者体内 CD~+_4 T 细胞存在过度增殖,可能与细胞周期蛋白和信号转导蛋白异常有密切关系。
Objective To investigate the proliferation of peripheral blood T lymphocyte subsets in asthmatic patients with remission and its molecular mechanism. Methods Peripheral blood samples from 45 patients with acute exacerbation of asthma (acute asthma group), patients with remission of asthma (asthma remission group) and normal controls (control group) were collected and cultured in vitro. Three groups of patients (P27 ~ (kipl), Cyclin D, E] in peripheral blood were detected by flow cytometry. The changes of cell cycle distribution, PI_3K and STAT_6 expression levels. Results (1) The distribution of G 0 / G 1 phase, the distribution of S phase, the ratio of G 2 / M phase and (S + G 2) / M phase in CD 3 + T lymphocytes in 3 groups were statistically significant (P <0.05). The proportion of G 0 / G 1 phase, the distribution of S phase and the ratio of (S + G 2) / M in CD 3 + T lymphocytes in 3 groups were statistically different (P <0.01). (2) The expressions of P27 kip1, Cyclin D and Cyclin E in CD 4 T lymphocytes in 3 groups were significantly different (Z values were 21.60, 25.22 and 21.52, (P <0.05). There was significant difference in P27 kipl expression among three groups of CD8 + T lymphocytes (Z value was 9.30, P <0.05) ) In PI3K and STAT6 protein in CD ~ + _4 T lymphocytes of three groups were significantly different (Z = 9.04 and 18.08, P <0.05 and 0.01, respectively). In the patients with remission of asthma, + _4 T cells were significantly increased compared with the normal controls, compared with patients with asthma attack in proliferative state CD ~ + _4, CD ~ + _8 T cells was significantly reduced. The PI3K signal transduction pathway may affect the proliferation and activation of CD8 +, CD4 + T cells by up-regulating CyclinD and / or CyclinE, while JAK1-STAT6 signaling may play a role by down-regulating P27Kip1. Conclusion The over-proliferation of CD ~ + _4 T cells in asthmatic patients may be related to the abnormality of cyclin and signal transduction proteins.