UBC13对支气管哮喘模型小鼠Th2、Th17型细胞极化的影响

Effect of UBC13 on the Polarization of Th2 and Th17 Cells in Asthma Model Mice

试验旨在探究UBC13蛋白对支气管哮喘小鼠体内Th2、Th17细胞极化的影响。18只BALB/c小鼠随机均分为3组:PBS组、哮喘模型(OVA)组和UBC13蛋白(UBC13)组,OVA组和UBC13组采用卵清蛋白(OVA)和脂多糖(LPS)进行致敏和雾化建立哮喘模型,其中UBC13组于每次雾化前0.5 h腹腔注射100μg UBC13蛋白。末次雾化激发24 h后处死小鼠,采集样品,通过HE染色观察肺脏切片病理学变化、PAS染色观察气道黏液的分泌,ELISA法检测血清IgE浓度和肺支气管肺泡灌洗液(BALF)上清中IL-4、IL-5和IL-17A的水平,实时荧光定量PCR法检测肺脏Th2型相关因子IL-4、IL-5、IL-13、IFN-γmRNA和Th17型细胞因子IL-1β、IL-6、IL-17A、IL-23 mRNA的相对表达量。结果显示,试验成功建立了哮喘模型,与PBS组相比,OVA组IgE浓度极显著上升(P<0.01),肺脏病理切片炎性细胞浸润和中性黏液分泌现象明显,肺脏中IL-1β、IL-4、IL-6、IL-13和IL-17A mRNA相对表达量均极显著升高(P<0.01),IL-5和IL-23 mRNA相对表达量显著升高(P<0.05),IFN-γmRNA相对表达量极显著下降(P<0.01),BALF中IL-4、IL-5和IL-17A的水平极显著上升(P<0.01);与OVA组相比,UBC13组IgE浓度显著降低(P<0.05),病理切片炎性细胞浸润现象减轻、黏液分泌减少,肺脏IL-1β、IL-4、IL-17A、IL-13和IL-23 mRNA相对表达量均显著下降(P<0.05),IFN-γmRNA相对表达量显著升高(P<0.05);IL-6 mRNA相对表达量极显著下降(P<0.01),BALF中IL-5的水平极显著下降(P<0.01),IL-4的水平显著下降(P<0.05),IL-17A的水平无显著差异(P>0.05)。由此可见,UBC13蛋白能下调Th2和Th17型细胞因子的表达量,影响哮喘模型Th2和Th17细胞的极化。

The aim of this study was to investigate the effect of UBC13 protein on the polarization of Th2 and Th17 cells in asthma model mice.18 BALB/c mice were randomly divided into PBS,asthma model(OVA)and UBC13 protein(UBC13)groups.The OVA and UBC13 groups were sensitized and aerosolized using ovalbumin(OVA)and lipopolysaccharide(LPS)to establish asthma model.Then 0.5 h before each atomization,UBC13 group was intraperitoneally injected with 100μg UBC13.After 24 h of the last atomization,all mice were killed and samples were taken.HE and PAS staining were used to observe the pathological changes of lung sections and the secretion of airway mucus,respectively.The IgE concentration in serum and the levels of IL-4,IL-5 and IL-17A in bronchoalveolar lavage fluid(BALF)were detected by ELISA,the relative expression of Th2(IL-4,IL-5,IL-13 and IFN-γ)and Th17(IL-1β,IL-6,IL-17A and IL-23)cytokines were detected by Real-time PCR.The results showed that the asthma model was successfully established.Compared with PBS group,the IgE content in OVA group increased extremely significantly(P<0.01),and the lung also occured the pathological changes.The relative expression of IL-1β,IL-4,IL-6,IL-13 and IL-17A mRNA increased extremely significantly in lung(P<0.01),the relative expression of IL-5 and IL-23 mRNA increased significantly(P<0.05),the relative expression of IFN-γmRNA declined extremely significantly(P<0.01),and the levels of IL-4,IL-5 and IL-17A in BALF increased extremely significantly(P<0.01).In addition,compared with OVA group,the serum IgE content declined significantly in UBC13 group(P<0.05).HE and PAS stainings results showed that inflammatory cell infiltration and mucus secretion decreased in pathological sections.The relative expression of IL-1β,IL-4,IL-17A,IL-13 and IL-23 mRNA declined significantly in lung(P<0.05),the relative expression of IFN-γmRNA increased significantly in lung(P<0.05),the relative expression of IL-6 mRNA declined significantly(P<0.01).The level of IL-4 and IL-5 in BALF decreased significantly(P<0.01;P<0.05),but the level of IL-17A was not significant difference(P>0.05).It indicated that UBC13 protein could downregulate the expression of Th2 and Th17 cytokines and inhibit the polarization of Th2 and Th17 cells in asthma model.