miR-125a-3p靶向FOXM1调控银屑病大鼠皮肤损伤和炎症反应的作用机制  

Mechanism of miR-125a-3p targeting FOXM1 in regulating skin injury and inflammatory response in psoriasis rats

作　　者：金曌 刘钟[1] 彭静 胡荣毅[1] 吴娟[1] 黄琴斯[1] 王飞 JIN Zhao;LIU Zhong;PENG Jing;HU Rongyi;WU Juan;HUANG Qinsi;WANG Fei(Department of Dermatology,Wuhan No.1 Hospital,Wuhan 430022,China;College of Acupuncture and Orthopedics,Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]武汉市第一医院皮肤科,武汉430022 [2]湖北中医药大学针灸骨伤学院,武汉430065

出　　处：《中国免疫学杂志》2024年第12期2531-2536,2542,共7页Chinese Journal of Immunology

基　　金：武汉市卫生健康委科研项目(WX21A13)。

摘　　要：目的:探究miR-125a-3p对银屑病大鼠皮肤损伤和炎症反应的影响及机制。方法:SD大鼠随机分为对照组、银屑病组、miR-NC组和miR-125a-3p组,对大鼠进行银屑病面积和严重性指数(PASI)评分、Baker评分;ELISA检测皮肤组织IL-6、IL-1β、TNF-α水平;qRT-PCR检测皮肤组织miR-125a-3p表达;qRT-PCR、Western blot检测皮肤组织叉头盒蛋白M1(FOXM1)mRNA和蛋白表达。分离培养银屑病大鼠角质形成细胞,双荧光素酶报告基因实验验证miR-125a-3p与FOXM1的靶向关系;分别抑制或过表达miR-125a-3p以及过表达miR-125a-3p基础上过表达FOXM1,检测细胞miR-125a-3p、FOXM1mRNA和蛋白表达以及细胞培养上清中IL-6、IL-1β、TNF-α水平,CCK-8法检测细胞活力,流式细胞术检测细胞凋亡率。结果:与对照组比较,银屑病组大鼠皮肤组织miR-125a-3p表达降低,PASI评分、Baker评分、IL-6、IL-1β、TNF-α水平以及FOXM1mRNA和蛋白表达升高(P<0.05);与银屑病组、miR-NC组比较,miR-125a-3p组大鼠皮肤组织miR-125a-3p表达升高,PASI评分、Baker评分、IL-6、IL-1β、TNF-α水平以及FOXM1 mRNA和蛋白表达降低(P<0.05)。miR-125a-3p与FOXM1存在靶向关系。抑制miR-125a-3p表达后,细胞FOXM1 mRNA和蛋白表达、细胞活力以及细胞培养上清中IL-6、IL-1β、TNF-α水平升高,细胞凋亡率降低(P<0.05),过表达miR-125a-3p则作用相反;过表达FOXM1可减弱过表达miR-125a-3p对细胞活力、凋亡率及炎症反应的影响。结论:miR-125a-3p在银屑病大鼠皮损组织中低表达,其过表达可能通过靶向下调FOXM1抑制角质形成细胞增殖并促进其凋亡,改善银屑病大鼠皮肤损伤,减轻炎症反应。Objective:To explore effect of miR-125a-3p on skin injury and inflammatory response in psoriasis rats and its mechanism.Methods:SD rats were randomly divided into control group,psoriasis group,miR-NC group and miR-125a-3p group.Psoriasis Area and Severity Index(PASI)score and Baker score were measured on rats;levels of IL-6,IL-1βand TNF-αin rat skin tissue were detected by ELISA;qRT-PCR was used to detect miR-125a-3p expression;mRNA and protein expressions of forkhead box protein M1(FOXM1)in rat skin tissue were detected by qRT-PCR and Western blot.Keratinocytes from psoriasis rats were isolated and cultured,targeting relationship between miR-125a-3p and FOXM1 was verified by dual-luciferase reporter gene assay.Inhibiting or overexpressing miR-125a-3p and FOXM1 was overexpressed on basis of overexpressing miR-125a-3p,respectively.miR-125a-3p,FOXM1 mRNA and protein expressions in cells and IL-6,IL-1β,TNF-αlevels in cell culture supernatants were detected,CCK-8 method was applied to detect cell viability,and flow cytometry was used to detect apoptosis rate.Results:Compared with control group,miR-125a-3p expression in skin tissue of rats in psoriasis group was decreased,PASI score,Baker score,IL-6,IL-1β,TNF-αlevels,and FOXM1 mRNA and protein expressions were increased(P<0.05);compared with psoriasis group and miR-NC group,expression of miR-125a-3p in skin tissue of rats in miR-125a-3p group was increased,PASI score,Baker score,IL-6,IL-1β,TNF-αlevels,and expressions of FOXM1 mRNA and protein were decreased(P<0.05).There was a targeting relationship between miR-125a-3p and FOXM1.After inhibiting miR-125a-3p expression,FOXM1 mRNA and protein expressions in cells,cell viability and IL-6,IL-1β,TNF-αlevels in cell culture supernatant were increased,and apoptosis rate was decreased(P<0.05),while overexpression of miR-125a-3p had opposite effect.Overexpression of FOXM1 attenuated effects of overexpression of miR-125a-3p on cell viability,apopto-sis rate and inflammatory response.Conclusion:miR-125a-3p is lowly exp

关 键 词：miR-125a-3p 叉头盒蛋白M1 银屑病 皮肤损伤 炎症反应 角质形成细胞 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]