miRNA和甲基化酶抑制剂在支气管哮喘中对Th细胞的影响

Effects of miRNA and methylase inhibitors on Th cells in bronchial asthma

目的探究支气管哮喘患者miR-126、miR-326和miR-155以及Th细胞相关因子的表达情况和5-氮杂胞苷(5-aza)对BA大鼠miR-126、miR-326和miR-155以及Th细胞相关因子的调控作用。方法QRT-PCR检测BA患者miR-126、miR-326和miRNA-155以及Th细胞相关因子mRNA的表达情况。建立BA大鼠模型,用不同浓度5-aza干预大鼠,PCR检测其miR-126、miR-326和miR-155以及Th细胞相关因子的表达。免疫组化检测IFN-γ、GATA3、ROR-γ、Foxp3蛋白表达。结果BA患者组miR-126(P<0.01)、miR-326和miR-155 mRNA表达高于正常组(P<0.05),IFN-γ和Foxp3 mRNA表达低于正常组(P<0.05),ROR-γmRNA表达高于正常组(P<0.01)。大鼠模型中,miR-126 mRNA在模型组的表达高于正常组(P<0.01);miR-126(P<0.05)、miR-326(P<0.01)和miR-155(P<0.05)mRNA在5-aza-L组中的表达低于模型组;免疫组化显示,IFN-γ在模型组的表达低于正常组(P<0.01),在5-aza-L组的表达高于模型组(P<0.01);GATA3和ROR-γ在模型组的表达高于正常组(P<0.01),在5-aza两组中的表达低于模型组(P<0.01);Foxp3在模型组的表达低于正常组(P<0.01),在5-aza两组的表达高于模型组(P<0.01)。结论甲基化酶抑制剂能下调miR-126、miR-326和miR-155的表达;甲基化酶抑制剂可以上调Foxp3以及下调GATA3和ROR-γ且双向调控IFN-γ的表达。

Objective To investigate the expression of miR-126,miR-326,miR-155,and Th cell-related factors in patients with bronchial asthma(BA)and the regulatory effects of the methylation enzyme inhibitor 5-azacytidin(5-aza)on miR-126,miR-326,miR-155,and Th cell-related factors in BA rats.Methods qRT-PCR was used to detect the mRNA expression of miR-126,miR-326,miRNA-155,and Th cell-associated factors in BA patients.The BA rat models were established,and rats were received different concentrations of 5-aza.The expression of miR-126,miR-326,miR-155,and Th cell-related genes in each group was detected by PCR.The expressions of IFN-γ,GATA3,ROR-γ,and Foxp3 were detected by immunohistochemistry.Results The expression of miR-126,miR-326,and miR-155 in the BA patient group was higher than in the normal group(P<0.05),while the mRNA expressions of IFN-γand Foxp3 were lower than those of normal group(P<0.05).The ROR-γmRNA in the BA patient group was higher than in the normal group(P<0.01).In the BA rat model,the expression of miR-126 mRNA in the model group was higher than that in the normal group(P<0.01);miR-126(P<0.05),miR-326(P<0.01),and miR-155(P<0.05)mRNA expression in 5-aza-L group was lower than that in model group.Immunohistochemical result showed that IFN-γexpression in the model group was lower than that in the normal group(P<0.01).IFN-γexpression in the 5-aza-L group was higher than that in the model group(P<0.01).GATA3 and ROR-γexpressions in the model group were higher than those in the normal group(P<0.01),and they were lower in the model group than that in the 5-aza two groups(P<0.01).Foxp3 expression in the model group was lower than that in the normal group(P<0.01),and its expression in the 5-aza two groups was higher than that in the model group(P<0.01).Conclusion Methylase inhibitors can down-regulate the expression of miR-126,miR-326 and miR-155.Methylase inhibitors can up-regulate Foxp3,down-regulate GATA3 and ROR-γ,and bidirectional regulate IFN-γexpression.