维生素D对慢性湿疹小鼠的改善及对miRNA155/SOCS1轴的调节作用研究

Improvement of Vitamin D on Mice with Chronic Eczema and Regulation of microRNA155/SOCS1 Axis

目的探讨维生素D对慢性湿疹小鼠的改善及对微小核糖核酸(miRNA)155/细胞因子信号转导抑制蛋白1(SOCS1)轴的调节作用。方法60只健康小鼠中随机取10只为A组,除外2只建模不成功,另48只右耳慢性湿疹模型小鼠以随机体质量排序法分为B组、C组、D组、E组,各12只。B组、D组、E组分别以5μg/kg miRNA155激动剂agomiR-155、10μg/kg 1,25-二羟维生素D3[1,25(OH)2D3]+5μg/kg agomiR-155混合液、10μg/kg 1,25(OH)2D3溶液均匀涂抹右耳,A组、C组以等量生理盐水涂抹右耳,1次/d,给药10 d。计算耳质量差、肿胀度;酶联免疫吸附测定(ELISA)法测定白细胞介素-4(IL-4)、IL-5、IL-2、干扰素-γ(IFN-γ);光镜下观察右耳组织病理形态学改变;实时聚合酶链反应(RT-PCR)法检测miRNA155、SOCS1 mRNA相对表达量;蛋白质印迹法(Western blotting)检测SOCS1蛋白相对表达量。结果与A组比较,B组耳质量差、肿胀度及IL-2、IFN-γ水平升高,IL-4、IL-5水平降低(P<0.05);与B组比较,C组、D组、E组耳质量差、肿胀度及IL-2、IFN-γ水平降低,IL-4、IL-5水平升高,且耳质量差、肿胀度及IL-2、IFN-γ,E组<D组<C组,IL-4、IL-5,C组<D组<E组(P<0.05);与B组、C组比较,D组、E组右耳组织表皮增厚、真皮炎性细胞浸润、水肿等异常改变减轻,其中E组减轻更明显;与A组比较,B组miRNA155 mRNA相对表达量升高,SOCS1 mRNA及蛋白相对表达量降低(P<0.05);与B组比较,C组、D组、E组miRNA155 mRNA相对表达量降低,SOCS1 mRNA及蛋白相对表达量升高,且miRNA155 mRNA相对表达量,E组<D组<C组,SOCS1 mRNA及蛋白相对表达量,C组<D组<E组(P<0.05)。结论维生素D对慢性湿疹小鼠病情有一定改善作用,机制可能与调节miRNA155/SOCS1轴有关。

Objective In order to investigate the effects of vitamin D on the improvement of chronic eczema in mice and the regulation of microRNA(miRNA)155/Suppressor of cytokine signaling1(SOCS1)axis.Methods Ten of 60 healthy mice were randomly selected as group A,and the other 48 right ear chronic eczema mice models were randomly divided into group B,group C,group D and group E with 12 mice in each group.Group B,group D and group E were evenly smeared with 5μg/kg miRNA155 agonist agomiR-155,10μg/kg 1,25-dihydroxyvitamin D3[1,25(OH)2D3]+5μg/kg agomiR-155 mixture and 10μg/kg 1,25(OH)2D3 solution,respectively.Group A and group C were smeared with the same amount of normal saline once a day for 10 days.The ear mass and swelling degree was calculated.The levels of interleukin-4(IL-4),IL-5,IL-2 and interferon-γ(IFN-γ)were detected by enzyme linked immunosorbent assay(ELISA).Hematoxylin eosin(HE)staining was used to detect the histopathology of the right ear.Real-time polymerse chain reaction(RT-PCR)was used to detect the relative expression of miRNA155 and SOCS1 mRNA.The relative expression of SOCS1 protein was detected by Western blotting.Results Compared with those in group A,the ear mass,swelling degree,IL-2 and IFN-γlevels in group B were increased,while IL-4 and IL-5 levels were decreased(P<0.05).Compared with group B,group C,group D and group E had lower ear mass,swelling degree and IL-2,IFN-γ levels,but higher IL-4,IL-5 levels,and the ear mass,swelling degree,IL-2 and IFN-γ,group E<group D<group C,IL-4,IL-5,group C<group D<group E(P<0.05).Compared with those in group B and group C,the abnormal changes of right ear tissue such as epidermal thickening,dermal cell infiltration and edema were alleviated in group D and group E,especially in group E.Compared with group A,the relative expression of miRNA155 mRNA in group B was increased,while the relative expression of SOCS1 mRNA and protein was decreased(P<0.05).Compared with group B,the relative expression of miRNA155 mRNA decreased and SOCS1 mRNA and protein increased in group C,D and E,and the relative expression level of miRNA155 mRNA,group E<group D<group C,SOCS1 mRNA and protein relative expression level,group C<group D<group E(P<0.05).Conclusion Vitamin D can improve the condition of mice with chronic eczema,and the mechanism may be related to the regulation of miRNA155/SOCS1 axis.