支气管哮喘病儿外周血单核细胞中LncRNA MEG3的表达及其与肺功能、免疫功能的相关性研究

Expression of LncRNA MEG3 in peripheral blood mononuclear cells of children with bronchial asthma and its correlation with lung function and immune function

目的探讨支气管哮喘病儿外周血单核细胞中长链非编码RNA母系表达基因3(LncRNA MEG3)的表达及其与肺功能和免疫功能的相关性。方法选取2017年10月至2018年9月安阳市妇幼保健院收治的支气管哮喘病儿60例为观察组,根据疾病严重程度分为轻度组(20例),中度组(20例),重度组(20例),同时选取同期体检的40例健康儿童作为对照组。均行肺功能检测,采用qRT-PCR检测各组外周血单核细胞中LncRNA MEG3表达量;血细胞分析仪检测嗜酸性粒细胞计数水平,ELISA法检测免疫球蛋白E(IgE)水平血清白细胞介素-4(IL-4)、白细胞介素-17A(IL-17A)、γ干扰素(IFN-γ)、转化生长因子-β(TGF-β)水平,比较各组肺功能、免疫功能指标、血清炎性介质及LncRNA MEG3水平差异,Pearson相关检验分析支气管哮喘病儿外周血单核细胞中LncRNA MEG3表达量与肺功能指标、免疫功能指标及炎性介质的相关性。多元线性回归分析LncRNA MEG3表达量与病情严重程度、肺功能、免疫功能的关联影响关系。结果重度组,中度组及轻度组LncRNA MEG3表达水平高于对照组[(0.30±0.07)、(0.42±0.06)、(0.67±0.10)比(1.01±0.13)](P<0.05),重度组LncRNA MEG3表达水平高于中度组及轻度组(P<0.05),中度组LncRNA MEG3表达水平高于轻度组(P<0.05);重度组,中度组及轻度组FEV1[(52.19±9.86)、(59.73±8.02)、(73.18±12.90)比(93.06±10.43)],FEV1/FVC[(65.23±9.65)、(72.05±5.28)、(80.12±8.12)比(93.25±10.90)]及PEF[(42.27±5.98)%、(59.94±5.67)%、(70.22±6.59)%比(86.52±3.57)%]低于对照组(P<0.05),重度组FEV1,FEV1/FVC及PEF低于中度组及轻度组(P<0.05),中度组FEV1,FEV1/FVC及PEF低于轻度组(P<0.05),重度组,中度组及低度组IgE[(110.25±13.32)U/mL、(84.92±12.98)U/mL、(56.61±11.21)U/mL比(23.21±5.81)U/mL]、嗜酸性粒细胞计数水平[(0.45±0.04)×10~9/L、(0.36±0.03)×10~9/L、(0.20±0.02)×10~9/L比(0.13±0.03)×10~9/L]、IL-4[(20.24±4.07)ng/L、(16.71±3.17)ng/L、(12.23±1.59)ng/L比(8.03±2.25)ng/L]、IL-17A[(119.51±8.52)ng/L、(100.89±6.28)ng/L、(85.74±9.05)ng/L比(52.67±13.24)ng/L]水平高于对照组(P<0.05),重度组IgE、嗜酸性粒细胞计数水平、IL-4、IL-17A水平高于中度组及轻度组(P<0.05),中度组IgE、嗜酸性粒细胞计数水平、IL-4、IL-17A水平高于轻度组(P<0.05),重度组、中度组及轻度组IFN-γ[(40.13±5.39)ng/L、(50.01±6.45)ng/L、(60.30±5.42)ng/L比(69.87±6.51)ng/L]、TGF-β[(709.74±36.53)ng/L、(836.77±42.64)ng/L、(1,002.35±53.34)ng/L比(1,940.88±224.39)ng/L]低于对照组(P<0.05),重度组IFN-γ、TGF-β低于中度组及轻度组(P<0.05),中度组IFN-γ、TGF-β低于轻度组(P<0.05),相关分析显示,支气管哮喘病儿LncRNA MEG3表达水平与IgE、IL-4、IL-17A呈负相关(P<0.05),与IFN-γ、TGF-β呈正相关(P<0.05),与FEV1,FEV1/FVC及PEF呈正相关(P<0.05)。多元线性回归显示,病情严重程度、肺功能、免疫功能与LncRNA MEG3表达量有密切的关联影响关系(P<0.05)。结论支气管哮喘病儿外周血单核细胞中LncRNA MEG3的表达量降低,并且与病情相关,其可能通过对免疫功能及炎症介质的调节参与支气管哮喘的发病及进展过程。

Objective To investigate the expression of long-chain noncoding RNA mother line expression gene 3(LncRNA MEG3)in peripheral blood mononuclear cells of children with bronchial asthma and its correlation with lung function and immune function.Methods Sixty children with bronchial asthma admitted to Anyang Maternal and Child Health Hospital from October 2017 to September 2018 were selected as the observation group.According to the severity of the disease,they were assigned into mild group(20 cases),moderate group(20 cases)and severe group(20 cases).At the same time,40 healthy children were selected as the control group.All the children had lung function test.The expression of LncRNA MEG3 in peripheral blood mononuclear cells was detected by qRTPCR.The eosinophil count was measured by blood cell analyzer.ELISA was used to detect the level of immunoglobulin E(IgE),and the levels of serum interleukin 4(IL-4),interleukin 17A(IL^-17A),interferon gamma(IFN–γ),transforming growth factor-beta(TGF–β).A comparison was made of the lung function,immune function,serum inflammatory mediators and the level of LncRNA MEG3 among the groups.Pearson correlation test was used to analyze the correlation between the expression of LncRNA MEG3 in peripheral blood mononuclear cells of asthmatic children and lung function,immune function and inflammatory mediators.Multiple linear regression was used to analyze the correlation between the expression of LncRNA MEG3 and the severity of the disease,lung function and immune function.Results The expression levels of LncRNA MEG3 in severe group,moderate group and mild group were higher than that in control group[(0.30±0.07)/(0.42±0.06)/(0.67±0.10)vs.(1.01±0.13)](P<0.05),and the expression of LncRNA MEG3 in severe group was higher than that in moderate group and mild group(P<0.05).And the expression of LncRNA MEG3 in moderate group was higher than that in mild group(P<0.05).FEV1[(52.19±9.86)/(59.73±8.02)/(73.18±12.90)vs.(93.06±10.43)],FEV1/FVC[(65.23±9.65)/(72.05±5.28)/(80.12±8.12)vs.(93.25±10.90)]and PEF[(42.27±5.98)%/(59.94±5.67)%/(70.22±6.59)%vs.(86.52±3.57)%]of severe group,moderate group and mild group were lower than those of control group(P<0.05).FEV1,FEV1/FVC and PEF of severe group were lower than those of moderate group and mild group(P<0.05),and FEV1,FEV1/FVC and PEF of moderate group were lower than those of mild group(P<0.05).The levels of IgE[(110.25±13.32)U/mL/(84.92±12.98)U/mL/(56.61±11.21)U/mL vs.(23.21±5.81)U/mL],eosinophil count[(0.45±0.04)×109/L/(0.36±0.03)×109/L/(0.20±0.02)×109/L vs.(0.13±0.03)×109/L],IL-4[(20.24±4.07)ng/L/(16.71±3.17)ng/L/(12.23±1.59)ng/L vs.(8.03±2.25)ng/L]and IL^-17A[(119.51±8.52)ng/L/(100.89±6.28)ng/L/(85.74±9.05)ng/L vs.(52.67±13.24)ng/L]in severe group,moderate group and mild group were higher than those in control group(P<0.05).The levels of IgE,eosinophil count,IL-4 and IL^-17A in severe group were higher than those in moderate group and mild group(P<0.05),and the levels of IgE,eosinophil count,IL-4 and IL^-17A in moderate group were higher than those in mild group(P<0.05).IFN–γ[(40.13±5.39)ng/L/(50.01±6.45)ng/L/(60.30±5.42)ng/L vs.(69.87±6.51)ng/L]and TGF-β[(709.74±36.53)ng/L/(836.77±42.64)ng/L/(1,002.35±53.34)ng/L vs.(1,940.88±224.39)ng/L]in severe group,moderate group and mild group were lower than those in control group(P<0.05).IFN-γand TGF–βin severe group were lower than those in moderate group and mild group(P<0.05),and those in moderate group were lower than those in mild group(P<0.05).Correlation analysis results showed that the expression level of LncRNA MEG3 was negatively correlated with IgE,IL-4 and IL^-17A(P<0.05),positively correlated with IFN-γand TGF-β(P<0.05),and positively correlated with FEV1,FEV1/FVC and PEF(P<0.05).Multiple linear regression showed that disease severity,lung function,immune function and LncRNA MEG3 expression were closely related(P<0.05).Conclusions The expression of LncRNA MEG3 in peripheral blood mononuclear cells of asthmatic children is decreased,which is related to the disease condition.It may participate in the pathogenesis and progression of asthma through the regulation of immune function and inflammatory mediators.