补肾固表方对复感儿外周血Th1和Th2型细胞因子表达失衡的调节作用

Regulative effects of Bushengubiao Recipe on unbalanced expressions of Th1 and Th2 cytokines from peripheral blood in children with recurrent respiratory tract infections

目的探讨补肾固表方对复感儿外周血T辅助细胞1(Th1)和T辅助细胞2(Th2)型细胞因子表达失衡的调节作用。方法采集20例复感儿治疗前后外周血,采用流式细胞仪分选CD4+T细胞,提取细胞总RNA,应用实时荧光定量PCR术,检测T-bet、GATA-3、干扰素(IFN-γ)、白细胞介素4(IL-4)基因表达量,并与10例健康对照组进行比较。结果复感儿T-bet mRNA和IFN-γmR-NA表达下降,GATA-3 mRNA和IL-4 mRNA表达升高,与正常组相比,有显著性差异(P<0.01或P<0.05)。补肾固表方治疗后T-bet mRNA上调(P>0.05),IFN-γmRNA上调(P<0.01),GATA-3mRNA和IL-4 mRNA下调(P<0.01),与正常组相比,无显著性差异(P>0.05),其中尤以GATA-3mRNA和IL-4mRNA改变程度明显。结论复感儿间歇期存在Th转录因子及细胞因子基因表达失衡,呈Th2优势应答模式。补肾固表方可使T-bet mRNA和IFN-γmRNA表达量增加,GATA-3mRNA和IL-4 mRNA表达量下降,从转录因子及细胞因子基因水平恢复Th平衡。

Objective To survey the regulative effects of Bushengubiao Recipe on the unbalanced expressions of Th1 and Th2 cytokines from peripheral blood in children with recurrent respiratory tract infections （RRTI）. Methods To select CD4^＋ T cells of peripheral blood in 20 patients of RRTI before and after treatment by flow cytometry, then abstract total RNA, determine the gene expressions of T-bet, GATA-3, IFN-γ and IL-4 by real time quantitative PCR, which were compared with those of 10 healthy controls. Results The expressions of T-bet mRNA and IFN-γ mRNA were lower, GATA-3 mRNA and IL-4 mRNA were higher in patients than in normal controls ,all with statistical significance （P 〈 0.01 or P 〈 0.05 ）. After Bushengubiao Recipe treatment, the expression of T-bet mRNA and IFN-γ mRNA upregulated （P 〉 0.05 ,P 〈 0.01 ）, GATA-3 mRNA and IL-4 mRNA down-regulated （ P 〈 0.01 ） without significant difference compared with healthy children （P 〉 0.05 ）. Conclusions The gene expression of Th transcription factor and cytokines is imbalanced in RRTI children in remission stage, the reaction mode in children of RRTI in remission stage is Th2 predominating. Bushengubiao Recipe can up-regulate T-bet mRNA and IFN-γ mRNA expression, down-regulate GATA-3 mRNA and IL4 mRNA expression, thus normalize the Th balance at the gene level of transcription factors and cytokines.