miR-34a在过敏性鼻炎小鼠中表达及治疗的作用研究  

Study on the expression and therapeutic effect of miR-34a in mice with allergic rhinitis

作　　者：谢伟伟 林秀联 龙绮[1] 孟光 刘硕 Xie Weiwei;Lin Xiulian;Long Qi;Meng Guang;Liu Shuo(Dept of Otorhinolaryngology Head and Neck Surgery,Affiliated to Xiangya Medical College, Central South University, Haikou Hospital, Haikou 570208)

机构地区：[1]中南大学湘雅医学院附属海口医院耳鼻咽喉头颈外科,海口570208

出　　处：《安徽医科大学学报》2022年第6期920-925,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81160129);海南省自然科学基金(编号:811145);海口市重点科技计划项目(编号:海科工信立﹝2012﹞72号)。

摘　　要：目的 探讨miR-34a在过敏性鼻炎小鼠中表达及治疗作用。方法 选取10只小鼠作为空白对照组,30只小鼠采用卵清蛋白构建小鼠过敏性鼻炎模型,随机分为模型组、miR-34a抑制剂组及阴性对照组(NC组)各10例,分别给予100μl生理盐水、100μl阴性转染质粒、100μl miR-34a抑制剂载体混合溶液尾部静脉注射。干预1周后观察小鼠行为学变化;酶联免疫吸附实验(ELISA)检测小鼠血清中卵清蛋白特异性IgE水平;qRT-PCR检测小鼠血清中白细胞介素(IL)-6、IL-8、IL-10、IL-35,基质金属蛋白酶(MMP)-2、MMP-9及血管细胞黏附分子(VCAM)-1 mRNA相对表达。流式细胞仪检测外周血CD3+、CD4+、CD8+T细胞含量。结果 模型小鼠血清miR-34a mRNA水平较空白组增加(P<0.05),与阴性对照组相比,miR-34a抑制剂干预后小鼠行为学总分降低(P<0.05),血清中卵清蛋白特异性IgE浓度降低(P<0.05),同时IL-6、IL-8、IL-10、MMP-2、MMP-9及VCAM-1 mRNA下降(P<0.05),IL-35上升(P<0.05),CD3+、CD4+/CD8+增加(P<0.05)。与空白对照组(NC组)相比,模型组IL-2、IFN-γ水平下降(P<0.05),与模型组相比,miR-34a抑制剂组IL-2、IFN-γ水平升高(P<0.05),miR-34a抑制剂组与NC组的上述方面比较,差异有统计学意义(P<0.05),模型组和NC组比较差异无统计学意义。结论 miR-34a在过敏性鼻炎小鼠中高表达,抑制miR-34a表达可减轻模型小鼠症状和炎症反应,提升免疫功能。Objective To investigate the expression and therapeutic effects of miR-34a in mice with allergic rhinitis.Methods 10 mice were selected as blank control group,and 30 mice were used to construct mouse allergic rhinitis model with ovalbumin.They were randomly divided into model group,miR-34a inhibitor group and negative control group(NC group)with 10 cases each.100μl of physiological saline,100μl of negative transfection plasmid,and 100μl of miR-34a inhibitor carrier mixed solution were injected into the tail vein respectively.The behavioral changes of mice after 1 week of intervention were observed;ELISA was used to detect the level of ovalbumin-specific IgE in mouse serum;qRT-PCR was used to detect interleukin(IL)-6,IL-8,IL-10,and IL-35,the relative expression of matrix metalloproteinase(MMP)-2,MMP-9 and vascular cell adhesion molecule(VCAM)-1mRNA.The content of CD3+,CD4+,CD8+T cells in peripheral blood was detected by flow cytometry.Results The serum level of miR-34a mRNA in model mice was significantly higher than that in the blank group(P<0.05).Compared with the negative control group,the total behavioral score of mice after miR-34a inhibitor intervention was significantly reduced(P<0.05).The concentration of ovalbumin-specific IgE was significantly reduced(P<0.05),while IL-6,IL-8,IL-10,MMP-2,MMP-9 and VCAM-1 mRNA were significantly reduced(P<0.05),IL-35 increased significantly(P<0.05),and CD3+,CD4+/CD8+increased significantly(P<0.05).Compared with the blank control group,the levels of IL-2 and IFN-γin the model group decreased(P<0.05);compared with the model group,the levels of IL-2 and IFN-γin the miR-34a inhibitor group and NC group increased(P<0.05);those indexes of miR-34a inhibitor group and NC group were compared,and there were statistically significant differences(P<0.05);there was no difference between the model group and NC group.Conclusion miR-34a is highly expressed in mice with allergic rhinitis.Inhibition of miR-34a expression can significantly reduce the symptoms and inflammation of model m

关 键 词：过敏性鼻炎 MIR-34A 抑制剂 炎症因子 T细胞 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]