miR-378a-5p靶向沉默FAIM促进银屑病皮下脂肪细胞促炎因子释放的研究  

MiR-378a-5p-targeted silencing of FAIM promotes pro-inflammatory factor release from psoriatic subcutaneous adipocytes

作　　者：彭静 尹婧[1] 夏萍[1] PENG Jing;YIN Jing;XIA Ping(Dermatology Department,Wuhan First Hospital,Wuhan 430000,China)

机构地区：[1]武汉市第一医院皮肤科,湖北武汉430000

出　　处：《安徽医学》2024年第10期1207-1214,共8页Anhui Medical Journal

基　　金：国家自然科学基金项目(编号:82304021)。

摘　　要：目的探究miR-378a-5p在银屑病皮下脂肪组织中的表达及其对银屑病影响机制。方法将SPF级4~6周龄雌性C57BL/6小鼠构建银屑病小鼠模型,其中Model组为银屑病小鼠,Control组为C57BL/6小鼠用凡士林乳膏。苏木精-伊红法(HE)染色法观察皮肤组织病理变化,实时荧光定量聚合酶链式反应(qPCR)检测皮下脂肪组织中miR-378a-5p、Fas细胞凋亡抑制分子(FAIM)的mRNA水平,蛋白质免疫印迹(WB)检测皮下脂肪组织中FAIM的蛋白水平。分离脂肪细胞进行后续实验。经双荧光素酶、qPCR和WB验证miR-378a-5p靶向FAIM。构建miR-378a-5p过表达和干扰质粒、及FAIM干扰质粒,脂肪细胞转染后,酶联免疫吸附测定(ELISA)检测培养基中炎症因子肿瘤坏死因子α(TNF-α)、白介素1(IL-1)和白介素6(IL-6)水平。结果模型组小鼠皮肤组织呈现明显银屑病病理形态。在银屑病小鼠模型脂肪组织中,miR-378a-5p的mRNA水平上调,FAIM的mRNA及蛋白水平均下调,差异均具有统计学意义(P<0.05)。双荧光素酶实验、qPCR和WB检测结果表明,FAIM是miR-378a-5p的下游靶点。与NC mimics组相比,miR-378a-5p mimics组脂肪细胞释放的炎症因子TNF、IL-1和IL-6水平显著上调(P<0.05);与NC inhibitors组,miR-378a-5p inhibi⁃tors脂肪细胞释放的炎症因子TNF-α、IL-1和IL-6水平下调(P<0.05)。与Control组相比,FAIM inhibitors组脂肪细胞释放的炎症因子TNF-α、IL-1和IL-6水平上调;而miR-378a-5p inhibitors+FAIM inhibitors可逆转这种现象(P<0.05)。结论miR-378a-5p靶向沉默FAIM促进银屑病皮下脂肪细胞促炎因子TNF-α、IL-1和IL-6的释放。Objective To investigate the expression of miR-378a-5p in subcutaneous adipose tissue of psoriasis and its impact on the pathogenesis of psoriasis.Methods A psoriasis mouse model was constructed from SPF grade 4~6 week old female C57BL/6 mice,where the model group was psoriasis mice and the control group was C57BL/6 mice with petroleum jelly cream.Hematoxylin eosin(HE)staining was used to observe the pathological changes in skin tissue.Real time fluorescence quantitative polymerase chain reaction(qPCR)was used to detect the mRNA levels of miR-378a-5p and Fas apoptosis inhibitory molecule(FAIM)in subcutaneous adipose tissue,and Western blot(WB)was used to detect the protein level of FAIM.Adipocytes were isolated for subsequent experiments.Dual-luciferase reporter assay,qPCR,and WB were used to validate the targeting relationship between miR-378a-5p and FAIM.Plasmids for overexpression and knockdown of miR-378a-5p,as well as FAIM knockdown,were constructed.After transfection into adipocytes,enzyme-linked immunosorbent assay(ELISA)was employed to measure the levels of inflammatory factors--tumor necrosis factorα(TNF-α),interleukin-1(IL-1),and interleukin-6(IL-6)in culture me⁃dium.Results The skin tissue of the model group mice showed obvious pathological morphology of psoriasis.In the adipose tissue of psoriasis mouse model,the mRNA level of miR-378a-5p was upregulated,while the mRNA and protein levels of FAIM were downregulated,and the dif⁃ferences were statistically significant(P<0.05).Dual-luciferase reporter assay,qPCR,and WB confirmed that FAIM was a downstream target of miR-378a-5p.Compared to the NC mimics group,the miR-378a-5p mimics group showed a significant increase in the release of inflamma⁃tory factors TNF-α,IL-1,and IL-6 from adipocytes(P<0.05).Compared to the NC inhibitors group,the miR-378a-5p inhibitors group showed a significant decrease in the release of TNF-α,IL-1,and IL-6(P<0.05).Compared to the control group,the FAIM inhibitors group showed a significant increase in the release of TN

关 键 词：银屑病 脂肪细胞 微小RNA-378a-5p Fas细胞凋亡抑制分子 炎症因子 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]