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目的观察PI3K信号通路对哮喘大鼠CD4~+白介素(IL)-17~+T细胞与CD4~+Foxp~(3+)调节性T细胞的影响,探讨该信号通路对哮喘大鼠CD4~+IL-17~+T细胞和CD4~+Foxp~(3+)T细胞失衡的调控作用。方法尼龙毛柱法分选对照组和哮喘组大鼠脾脏T细胞,对照组(A组)分为2个亚组:空白对照组(A1),PI3K抑制剂LY294002 20μmol/L对照组(A2);哮喘组(B组)分为4个亚组:哮喘组(B1),5μmol/L PI3K抑制剂LY294002组(B2),10μmol/L PI3K抑制剂LY294002组(B3),20μmol/L PI3K抑制剂LY294002组(B4),采用流式细胞仪检测CD4~+IL-17~+T细胞和CD4~+Foxp~(3+)T细胞,逆转录-聚合酶链反应检测特异性转录因子表达。结果CD4~+Foxp~(3+)T细胞的数量B1组较A1组降低(P<0.01);CD4~+IL-17~+T细胞的数量B1组较A1组升高(P<0.01)。IL-17水平和ROR-γt mRNA表达B1组较A1组增高(P<0.01);而IL-10水平和Foxp3 mRNA表达B1组低于A1组(P<0.01)。ROR-γt mRNA/Foxp3 mRNA比值与IL-17水平呈正相关(r=0.8726,P<0.01);ROR-γt/Foxp3 mRNA比值与IL-10呈负相关(r=-0.8504,P<0.01)。结论 PI3K信号通路可能通过调控CD4~+IL-17~+T细胞/CD4~+Foxp~(3+)T细胞的失衡而参与哮喘发病过程。
Objective To investigate the effects of PI3K signaling pathway on CD4 ~ + IL-17 ~ + T cells and CD4 ~ + Foxp ~ (3 +) regulatory T cells in asthmatic rats. Regulation of IL-17 ~ + T cell and CD4 ~ + Foxp ~ (3 +) T cell imbalance. Methods T lymphocytes from control group and asthmatic rat were separated by nylon hair column and divided into two groups: control group (A1), PI3K inhibitor LY294002 (20μmol / L), control group (A2) Asthma group (B1), 5μmol / L PI3K inhibitor LY294002 group (B2), 10μmol / L PI3K inhibitor LY294002 group (B3), 20μmol / L PI3K inhibitor LY294002 group (B4). Flow cytometry was used to detect CD4 ~ + IL-17 ~ + T cells and CD4 ~ + Foxp ~ (3+) T cells. Reverse transcriptase polymerase chain reaction was used to detect the expression of specific transcription factors. Results The number of CD4 ~ + Foxp ~ (3 +) T cells in B1 group was lower than that in A1 group (P <0.01). The number of CD4 ~ + IL-17 ~ + T cells in B1 group was higher than that in A1 group . The level of IL-17 and the expression of ROR-γt mRNA in group B1 were higher than those in group A1 (P <0.01), while the levels of IL-10 and Foxp3 mRNA in group B1 were lower than those in group A1 (P <0.01). The ratio of ROR-γt mRNA to Foxp3 mRNA was positively correlated with IL-17 (r = 0.8726, P <0.01). The ratio of ROR-γt / Foxp3 mRNA was negatively correlated with IL-10 (r = -0.8504, P <0.01). Conclusion The PI3K signaling pathway may be involved in the pathogenesis of asthma by regulating the imbalance of CD4 ~ + IL-17 ~ + T cells / CD4 ~ + Foxp ~ (3 +) T cells.