摘要
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 evaluate the changes of CD4^＋IL- 17^＋T（ Th17） and CD4^＋Foxp^3＋regulatory T（ Treg）cells in peripheral blood,and explore the regulatory effect of PI3 K signal pathway on CD4^＋Foxp^3＋regulatory T cells and CD4^＋IL- 17^＋T cells in murine asthma model. Methods Wistar mice were used in the study. The mice were randomly divided into control group and asthma group. The mice in asthma group were intraperitoneally injected with the mixture of ovalbumin（ OVA） / Al（ OH）3and then activated by exposure of the animals to OVA atomization. The T cells in the spleens from Wistar mice were isolated by the method of nylon wool column,and the T cells were divided into different subgroups： the T cells from control group were divided into normal control group（ A1） and PI3 K inhibitor- treated group（ A2）; the T cells from asthma group were divided into asthma group（ B1）,5 μmol / L PI3 K inhibitor- treated group（ B2）,10 μmol / L PI3 K inhibitor- treated group（ B3） and 20 μmol / L PI3 K inhibitor- treated group（ B4）.The number of CD4^＋IL- 17^＋T cells and CD4^＋Foxp^3＋T cells were detected by flow cytometry. The transcription levels of Foxp3 and ROR- γt were determined by RT- PCR. Results The number of CD4^＋Foxp^3＋Treg in group B1 was significantly lower than that in group A1,the levels of IL- 17 and mRNA expression of ROR- γt in B1 group were much higher than those in A1 group. However,the levels of IL- 10 and mRNA expression of Foxp3 in B1 group were significantly lower than those in group A1,the ratio of ROR- γt mRNA / Foxp3 mRNA was positively correlated with the level of IL- 17 and was negatively correlated with the level of IL- 10. Conclusion PI3 K signal pathway plays an important role in regulating the imbalance of CD4^＋IL- 17^＋T / CD4^＋Foxp^3＋T in asthma.
出处
《医药论坛杂志》
2016年第8期22-24,共3页
Journal of Medical Forum
