摘要
目的探讨罗格列酮对支气管哮喘(简称哮喘)急性发作期患者 T 淋巴细胞转录因子T-bet 与转录因子 GATA-3表达的调控、对 Th1/Th2细胞因子的影响及与地塞米松(DXM)作用机制的异同。方法哮喘急性发作期患者(A 组)10例及健康自愿者(B 组)10名。分离、纯化、培养外周血T淋巴细胞,根据实验要求将两组分为3、24 h 组,再根据不同的干预分为:未干预对照组(A_1组、B_1组)、罗格列酮干预组(A_2组、B_2组)及 DXM 干预组(A_3组、B_3组);用酶联免疫吸附(ELISA)法检测T淋巴细胞培养上清液中γ干扰素(IFN-γ)及白细胞介素4(IL-4)的水平,用逆转录-聚合酶链反应(RT-PCR)法检测 T 淋巴细胞 T-bet mRNA、GATA-3 mRNA 的表达水平。结果 A 组3、24 h T 淋巴细胞培养上清液中 IFN-γ、IFN-γ/IL-4和 T 淋巴细胞的 T-bet mRNA、T-bet mRNA/GATA-3 mRNA 水平分别为357±31、783±47、5.5±1.0、8.4±1.5、18.7±3.7、11.9±2.9、1.20±0.11、0.290±0.020,与 B组3、24 h(659±41、1394±120、11.5±3.0、17.4±4.0、29.0±4.0、18.9±4.0、3.82±0.81、0.870±0.090)比较差异有统计学意义(t 值分别为18.59、5.95、14.87、6.25、6.06、4.63、10.23、18.67,P 均<0.05),A 组3、24 h IL-4、GATA-3 mRNA 水平分别为65±6、96±9、16.4±4.2、41±6,与 B 组3、24 h(57±5、83±7、7.8±2.2、23±4)比较差异有统计学意义(t 值分别为3.19、3.90、5.77、7.76,P 均<0.05);A、B 组 T 淋巴细胞培养上清液中 IFN-γ,、IL-4、IFN-γ/IL-4在3 h 水平分别为357±31、65±6、5.5±1.0、659±41、57±5、11.5±3.0,与24 h(783±47、96±9、8.4±1.5、1394±120、83±7、17.4±4.0)水平比较差异有统计学意义(t 值分别为23.8、9.48、5.03、18.21、9.01、3.53,P 均<0.05);A、B组 T 淋巴细胞 T-bet mRNA 和 T-bet mRNA/GATA-3 mRNA 在3 h 时分别为18.7±3.7、1.20±0.11、29.0±4.0、3.82±0.81,与24 h(11.9±2.9、0.290±0.020、18.9±4.0、0.870±0.090)水平比较差异有统计学意义(t 值分别为4.62、5.66、29.67、11.5,P 均<0.05),但 A、B 组 GATA-3 mRNA 在3 h(16.4±4.2、7.8±2.2)的水平,分别低于24 h(41±6、23±4)水平(t 值分别为10.70、10.11,P 均<0.05);A 组 T 淋巴细胞 T-bet mRNA 表达与 T 淋巴细胞培养上清液中 IFN-γ呈正相关(r=0.581,P<0.05),与 IL-4呈负相关(r=-0.493,P<0.05);A 组 GATA-3 mRNA 表达与 IFN-γ呈负相关(r=-0.501,P<0.05),与 IL-4呈正相关(r=0.579,P<0.05);A 组 T-bet mRNA/GATA-3 mRNA 比值与IFN-γ/IL-4比值呈正相关(r=0.696,P<0.05)。结论罗格列酮通过影响哮喘急性发作期患者 T淋巴细胞 T-bet 和 GATA-3的表达,从而调控 IFN-γ和 IL-4的平衡。
Objective To investigate the effect of rosiglitazone on the expression of T-bet/GATA-3 in peripheral blood T lymphocytes and eytokine derived from Th1/Th2 in patients with acute asthma and to compare its mechanism with that of dexamethasone （DXM）. Methods Peripheral blood T lymphocytes from 10 patients with acute asthma （A group, A） and 10 healthy volunteers（B group,B） were isolated, purifieated and cultured, and were divided into 2 groups（3 h and 24 h） based on the treatment time, and each group was further divided, on the basis of the treatment given, into 3 sub-groups respectively： control group （ A1 and B1 ）, rosiglitazone treated group （ A2 and B2 ） and DXM treated group （ A3 and B3 ）. The levels of IFN-γ and IL-4 in the culture supernatant were detected by Enzyme-linked immunoadsorbent assay （ELISA） and the expression levels of T-bet mRNA/GATA-3 mRNA in T lymphocytes was detected by Reverse Transcription-Polymerase Chain Reaction （RT-PCR）. Results The levels of INF-γ and INF-γ/ IL-4 in the culture supernatant of T lymphocytes and T-bet mRNA and T-bet mRNA/GATA-3 mRNA in T lymphocytes from the A at 3 h and 24 h were 357±31,783 ± 47,5.5 ± 1.0,8. 4 ± 1.5,18.7 ± 3.7,11.9 ± 2. 9,1.20 ±0. 11,0. 290 ±0. 020, and those from the B at 3 h and 24 h were 659 ±41,1394 ± 120,11.5 ± 3.0,17.4±4.0,29.0 ±4.0, 18.9 ±4.0,3.82 ±0.81,0.870 ±0.090, the differences were significant between these groups （t = 18.59, 5.95, 14.87, 6.25, 6.06, 4.63, 10.23, 18.67, all P 〈 0.05, respectively）. The levels of IL-4 in the culture supernatant of T lymphocytes and GATA-3 mRNA in T lymphocytes from the A at 3 h and 24 h were 65 ±6,96 ±9,16.4 ±4. 2,41 ±6, and those from the B at 3 h and 24 h were 57 ±5,83 ±7,7. 8 ±2. 2, 23 ±4, the differences were significant between these groups（t = 3. 19,3.90,5.77,7.76, all P 〈 0. 05, respectively）. The levels of IFN-γ, IL-4 and IFN-γ/IL-4 in the culture supernatant of T lymphocytes from the A and B at 3 h were 357 ± 31,65 ± 6,5.5 ± 1.0, 659 ± 41, 57 ± 5,11.5 ± 3.0, and those from the A and B at 24 h were 783 ± 47,96 ± 9,8.4 ± 1.5,1394 ± 120,83 ± 7,17. 4 ± 4. 0, The differences were significant between these groups （ t = 23.8,9.48,5.03,18.21,9. 01, 3.53,all P 〈 0.05, respectively）.The levels of T-bet mRNA and T-bet mRNA/GATA-3 mRNA in T lymphocytes from the A and B at 3 h were 18.7 ± 3.7,1.20 ± 0. 11,29.0 ± 4. 0,3.82 ± 0. 81, and those from the A and B at 24 h were 11.9 ±2.9,0. 290±0. 020,18. 9 ±4. 0,0. 870±0. 090, the differences were significant between these groups （ t = 4. 62,5.66,29. 67,11.5, all P 〈 0. 05, respectively）. The levels of GATA-3 mRNA in T lymphocytes from the A and B at 3 h were 16.4 ±4. 2,7. 8 ±2. 2, it was lower than those from the A and B at 24 h （41 ±6,23 ±4, t = 10. 70,10. 11 ,all P 〈0. 05, respectively）. The levels of T-bet mRNA from the A showed a positive correlation with the level of IFN-γ （r =0. 581 ,P 〈0. 05）, and a negative correlation with the level of IL-4 （ r = - 0. 493, P 〈 0. 05 ） ; The levels of GATA-3 mRNA from the A showed a negative correlation with the level of IFN-γ （r = -0. 501 ,P 〈0. 05）, and a positive correlation with the level of IL-4 （ r = 0. 579, P 〈 0. 05 ） ; The ratio of T-bet mRNA to GATA-3 mRNA from the A showed a positive correlation with the ratio of IFN-γ to IL-4 （ r = 0. 696, P 〈 0. 05 ） . Conclusion Rosiglitazone could regulate the balance of IFN-γ and IL-4 through effect on the expression of T-bet mRNA and GATA-3 mRNA.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
2007年第2期116-120,共5页
Chinese Journal of Tuberculosis and Respiratory Diseases
基金
湖南省科技厅基金(05fj3035)
