摘要
目的:探讨哮喘患者PBMCs中转录因子T-bet/GATA-3比率与Th1/Th2细胞失衡的关系及体外CpG干预后T-bet/GATA3比率的变化及其对Th1/Th2细胞平衡的调节作用。方法:用RT-PCR法测定30例发作期哮喘患者(哮喘组)及20例慢性阻塞性肺病患者(COPD组)及20例正常人(对照组)PBMCs中T-bet mRNA、GATA-3 mRNA及TLR9 mRNA的表达强度,用ELISA法测定血浆IL-4、IL-5、IL-13和IFNγ的表达水平,以观察哮喘患者PBMCs中转录因子T-bet/GATA-3比率与Th1/Th2细胞失衡的关系。将20例发作期哮喘患者的PBMCs分为2部分,一份加入CpG ODN和PHA(CpG组),一份仅加入PHA(对照组),培养48h后分别收集培养液和细胞,采用RT-PCR法测定细胞中T-bet mRNA、GATA-3 mRNA及TLR9 mRNA的表达强度,ELISA法测定培养液中IL-4、IL-5、IL-13和IFN-γ的表达水平。结果:哮喘患者PBMCs中T-bet/GATA-3的比率显著低于COPD患者和正常人,IL-4、IL-5、IL-13的水平显著高于COPD患者和正常人,与T-bet/GATA-3的比率呈负相关,而IFN-γ的水平显著降低,与T-bet/GATA-3的比率呈正相关。哮喘组TLR9的表达强度显著弱于COPD患者和正常人。CpG干预组细胞培养液中IL-4、IL-5和IL-13的水平分别显著低于对照组,IFN-γ的水平显著高于对照组。CpG干预组细胞中T-bet mRNA和TLR9 mRNA的表达强度显著强于对照组,GATA-3 mRNA的表达强度显著低于对照组;T-bet/GATA-3的比率显著高于对照组。结论:哮喘患者T-bet/GATA-3的比率降低,可以作为评价Th1/Th2细胞失衡的精确指标。CpG ODN可以上调T-bet的表达,下调GATA-3的表达,从而上调T-bet/GATA-3的比率,逆转Th1/Th2细胞失衡,是一种很有前景的哮喘治疗手段。
AIM： The aim of this study is to investigate the relationship between the ratio of transcription factors T- bet/GATA- 3 in peripheral blood mononuclear cells （PBMCs） and the imbalance of Th1/Th2 cytokines from patients with asthma, and to examine whether and how CpG ODN intervention would restore the imbalanced Th1/Th2 profile in asthma patients. METHODS： Three groups of subjects with asthma （asthma group）, chronic obstructive pulmonary disease （COPD group）, and normal controls （control group） were included in this study. T -bet, GATA -3 and TLR9 mRNA levels were measured by reverse transcription -polymerase chain reaction （RT -PCR）. Th1 cytokine IFN -γ and Th2 cytokines IL -4, IL -5 and IL - 13 were measured by enzyme -linked immunoabsorbent assay （ELISA） in asthma group. Cultured PBMCs from asthma patients with CpG ODN incubation were collected for measuring T - bet, GATA - 3 mRNA and TLR9 mRNA. RESULTS ： The ratio of T - bet/GATA - 3 in asthma group was significantly lower than that in the COPD group and control group. The serum levels of IL -4, IL -5 and IL - 13 had positive correlation with the ratio of T - bet/GATA- 3 in asthmatic patients, and the lower serum level of IFN -γ had negative correlation with the ratio. The TLR9 mRNA expression in asthma was significantly attenuated than that in COPD group and control group. Consequently, the ratio of T - bet/GATA - 3 in CpG group was significantly enhanced than that in control group. CONCLUSION ： These results indicate that the ratio of T - bet/GATA - 3 in asthma is decreased and could represent the imbalance of Th1/Th2 cells. CpG ODN influences the ratio of T - bet/GATA - 3, upregulating the T - bet mRNA expression and downregulating the GATA - 3 mRNA expression, which is able to reverse the imbalance of Th1/Th2 cells. CpG - ODN may be a promising therapeutic target for asthma.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第9期1776-1781,共6页
Chinese Journal of Pathophysiology
基金
山东省卫生厅科研基金(No.2005HW058)
