生血合剂对慢性再生障碍性贫血患者T-bet/GATA-3、相关信号分子、细胞因子表达及Th<sub>1</sub>/Th<sub>2</sub>的影响

Influence of Shengxue Mixture on the Expression of T-bet/GATA-3,Their Relevant Signal Transduction Molecules,Cytokines,and Th₁/Th₂ Balance in Patients with Chronic Aplastic Anemia

免费下载全文查看/发表评论下载PDF阅读器

DOI：

中文关键词: 慢性再生障碍性贫血表达于T细胞的T盒转录因子信号传导和转录激活因子4 1型辅助性T细胞/2型辅助性T细胞失衡生血合剂

英文关键词:chronic aplastic anemia T-bet signal transducers and activators of transcription 4 disequilibrium of Th₁/Th₂ Shengxue Mixture

基金项目:上海市教委科研基金资助项目(No.07cz031);上海市卫生局科研基金资助项目(No.2007Y64)

作者	单位
李峻	江苏省中医院(南京中医药大学附属医院)血液科
周永明	上海中医药大学附属岳阳中西医结合医院血液科
胡明辉	上海中医药大学附属岳阳中西医结合医院血液科
孙伟玲	上海中医药大学附属岳阳中西医结合医院血液科
薛志忠	上海中医药大学附属岳阳中西医结合医院检验科

摘要点击次数: 1286

全文下载次数: 640

中文摘要:

目的研究转录因子T-bet（T-box expressed in T cells）、GATA-3（GATA binding protein3）及相关信号通路在慢性再生障碍性贫血（chronic aplastic anemia,CAA）免疫发病中的作用,从辅助性T（helper T,Th）细胞失衡、转录因子及相关信号通路水平探讨生血合剂治疗CAA的免疫调控机制。方法 40例CAA患者均来自上海中医药大学附属岳阳医院,随机分为治疗组20例和对照组20例,选取体检健康者20名为正常组。治疗组患者辨证分为脾肾阳虚型和脾肾阴虚型,脾肾阳虚型患者服用生血合剂1号,脾肾阴虚型患者服用生血合剂2号,对照组服用环孢菌素A（CsA）,采用实时荧光定量PCR（Realtime FQ-PCR）检测CAA患者治疗前后外周血单个核细胞（peripheral blood mononuclear cell,PBMNC）、T-bet、GATA-3及信号转导子及转录激活因子4（signal transducer and activator of transcription 4,STAT4）、STAT6 mRNA表达,采用流式细胞术、酶联免疫吸附法（ELISA）检测CAA患者治疗前后PBMNC Th₁、Th₂比例及PBMNC培养上清IFN-γ、IL-12、IL-4水平。结果 CAA患者PBMNC T-bet、STAT4 mRNA表达、T-bet/GATA-3比值、Th₁比例、Th₁/Th₂比值、PBMNC培养上清IFN-γ、IL-12表达均明显高于正常组（P<0.01）,采用生血合剂或CsA治疗后,患者T-bet、STAT4表达、T-bet/GATA-3比值、Th₁比例、IFN-γ、IL-12表达均有所下降（P<0.01）,但T-bet、STAT4、T-bet/GATA-3比值、Th₁比例、IFN-γ表达仍未达到正常水平,治疗组与对照组比较差异无统计学意义（P>0.05）,而GATA-3、STAT6 mRNA表达、Th₂比例、IL-4表达治疗前后与正常组比较差异均无统计学意义（P>0.05）。结论 IFN-γ/T-bet、IL-12/STAT4通路的异常活化,及Th₁/Th₂平衡向Th₁型偏移,在CAA免疫异常的发病过程中起关键的作用。生血合剂和CsA能通过下调IFN-γ/T-bet、IL-12/STAT4通路的异常活化,并纠正Th₁过度极化,从而减轻CAA异常亢进的细胞免疫,解除造血抑制。

英文摘要:

Objective To study the actions of transcription factors,T-bet and GATA-3,and their relevant signal transduction pathways on the immune-related pathogenesis with chronic aplastic anemia（CAA）,and to investigate the immunological regulation mechanism of Shengxue Mixture（SXM） in regulating levels of Th cell imbalance,transcriptional factor and relevant signal pathways.Methods All CAA patients selected from Yueyang Hospital of Shanghai University of traditional Chinese medicine were equally randomized into the treated group and the control group,20 patients in each group,and 20 healthy persons were selectod as normal group, the former was treated with SXM according to patients’ syndrome patterns,namely,SXM-1 was given to patients of Pi-Shen yang-deficiency pattern,and SXM-2 to those of Pi-Shen yin-deficiency pattern.Patients in the control group were treated with cyclosporin A（CsA）.The mRNA expressions of T-bet,GATA-3,signal transducers and activators of transcription 4（STAT4） and 6（STAT6） in peripheral blood mononuclear cell（PBMNC） of patients were determined using real-time fluorescent quantitation polymerase chain reaction before and after treatment, meantime,the Th₁/Th₂ proportion in peripheral blood,and levels of IFN-γ,IL-12 and IL-4 in PBMNC-cultured supernatant were detected by flow cytometry and enzyme linked immunosorbent assay.Results The mRNA expressions of PBMNC T-bet and STAT4,ratios of T-bet/GATA-3,Th,proportion and Th₁/Th₂ ratio,levels of IFN-γand IL-12 in PBMNC-cultured supernatant were all significantly higher in CAA patients than in healthy controls （P<0.01）,which were lowered after treatment but didn’t reach the normal range（all P<0.01）,excepting for IL-12 level.Comparisons of the changes between the two treated groups showed insignificant difference（P>0.05）.While the difference between patients and healthy persons in terms of GATA-3,STAT6,Th₂ proportion, and IL-4 were insignificant（P>0.05）,either before or after treatment.Conclusions Abnormal activation of IFN-γ/T-bet and IL-12/ STAT4 pathways,as well as Th₁/Th₂ balance deviating to Th₁ excursion play vital roles in the immunological pathogenesis of CAA.SXM and CsA could lower the aforesaid abnormal activation and correct Th₁ hyper-polarization,so as to alleviate the over-activated cell-mediated immunity to eliminate hematopoietic depression in CAA patients.

关闭