血清外泌体miR-30d-5p靶向RHOB在三氯乙烯药疹样皮炎中的作用  

Role of serum exosomal miR-30d-5p targeting RHOB in occupational medicamentosa-like dermatitis due to trichloroethylene

作　　者：蔡曙阳 王慧[2,3] 张雪松[1] 左绪磊 马金茹 洪依婷 吴奇峰 朱启星[1,2,3] Cai Shuyang;Wang Hui;Zhang Xuesong;Zuo Xulei;Ma Jinru;Hong Yiting;Wu Qifeng;Zhu Qixing(Dept of Occupational Hygiene and Environmental Hygiene,School of Public Health,Anhui Medical University,Hefei 230032;Dept of Dermatology,the First Affiliated Hospital of Anhui Medical University,Hefei 230022;Key Laboratory of Dermatology,Ministry of Education,the First Affiliated Hospital of Anhui Medical University,Hefei 230022;Guangdong Province Hospital for Occupational Disease Prevention and Treatment,Guangzhou 510000)

机构地区：[1]安徽医科大学公共卫生学院职业卫生与环境卫生学系,合肥230032 [2]安徽医科大学第一附属医院皮肤病研究所,合肥230022 [3]安徽医科大学国家教育部皮肤病学重点实验室,合肥230022 [4]广东省职业病防治院,广州510000

出　　处：《安徽医科大学学报》2023年第5期718-723,730,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82173494、81874259)。

摘　　要：目的检测三氯乙烯药疹样皮炎(OMDT)患者血清外泌体中miR-30d-5p的表达水平及其与肝功能的相关性,再对miR-30d-5p进行生信分析并验证下游靶基因RHOB。方法收集6例OMDT患者和6例健康对照者血清样本提取血清外泌体。透射电镜、纳米粒子跟踪分析以及Western blot法对外泌体进行鉴定后提取miRNA,检测其中miR-30d-5p表达水平并与肝功能作相关性分析,用miRWalk及miRBD数据库预测miR-30d-5p的靶基因,进行功能聚类和途径富集分析,最后通过双荧光素酶报告基因实验验证靶基因RHOB。结果6例OMDT患者在发病高峰期时血清外泌体中miR-30d-5p表达水平明显降低(P<0.05),并且与肝功能指标丙氨酸氨基转移酶、天门冬氨酸氨基转移酶、γ-谷氨酰胺基转移酶水平均呈负相关(相关系数分别为:r_(s)=-0.943,P=0.005;r_(s)=-0.886,P=0.019;r_(s)=-0.886,P=0.019)。生信分析及双荧光素酶报告基因实验显示RHOB是miR-30d-5p的直接靶基因。结论OMDT患者血清外泌体miR-30d-5p表达降低,与肝功能指标呈负相关,并且可能通过靶向RHOB参与三氯乙烯所致肝损伤过程。Objective To detect the expression of serum exosomal miR-30d-5p in occupational dermatitis medicamentosa-like of trichloroethylene(OMDT)patients and its correlation with liver function,then perform bioinformatics analysis and verify the target gene.Methods Serum exosomes were extracted from 6 OMDT patients and 6 healthy controls,and miRNA was extracted from exosomes which were identified by transmission electron microscopy,nanoparticle tracking analysis and western blotting.The expression of serum exosomal miR-30d-5p was detected by real-time fluorescence quantitative PCR,then the correlation with liver function was analyzed.The target genes of miR-30d-5p were predicted by the miRWalk and miRBD databases.Gene ontology analysis and KEGG pathways analysis were performed.Finally,RHOB was verified by Dual-luciferase reporter assay.Results The expression of serum exosomal miR-30d-5p significantly decreased in 6 OMDT patients at the peak of the disease(P<0.05),and it was negatively correlated with the level of AST,ALT and GGT(correlation coefficient:r_(s)=-0.943,P=0.005;r_(s)=-0.886,P=0.019;r_(s)=-0.886,P=0.019).Bioinformatics analysis and dual-luciferase reporter assay showed that RHOB was the target gene of miR-30d-5p.Conclusion The expression of serum exosomal miR-30d-5p decreases in OMDT patients,which is negatively correlated with the level of liver function,and the target gene RHOB may be involved in the process of liver injury induced by trichloroethylene.

关 键 词：三氯乙烯 外泌体 miR-30d-5p RHOB 

分 类 号：R135.7[医药卫生—劳动卫生]