羧胺三唑通过下调细胞因子和抗菌肽S100A7的表达改善实验性银屑病

Carboxyamidotriazole ameliorates experimental psoriasis via downregulating the expressions of cytokines and antimicrobial peptide S100A7

应用咪喹莫特(imiquimod,IMQ)诱导的银屑病小鼠模型以及五联因子(a mixture of five proinflammatory cytokines,M5)诱导的银屑病角质形成细胞模型,评价羧胺三唑(carboxyamidotriazole,CAI)对银屑病的治疗效果和可能机制。银屑病皮损面积和疾病严重程度(psoriasis area and severity index,PASI)评分评估小鼠皮损严重程度,苏木素-伊红(hematoxylin-eosinstaining,HE)染色观察皮肤病理学变化,酶联免疫吸附法(enzyme-linked immunosorbentassay,ELISA)检测皮肤中细胞因子水平,转录组测序分析差异表达基因,实时荧光定量PCR(real-time quantitative PCR,qPCR)检测mRNA表达。动物实验经中国医学科学院基础医学研究所实验动物管理与伦理委员批准(批准号:ACUC-A02-2022-115)。结果显示,与IMQ组相比,CAI显著改善小鼠银屑病样皮损,降低PASI评分,减轻皮损病理变化和评分,减少皮损处促炎细胞因子水平,转录组分析结果显示CAI可能通过调节角质形成细胞及其介导的表皮角化而发挥作用,qPCR验证发现CAI能够下调差异表达基因S100a7;进一步使用M5刺激HaCaT细胞构建银屑病角质形成细胞模型,CAI能够抑制M5诱导的S100a7以及细胞因子的mRNA水平。综上,CAI可能对银屑病具有良好治疗作用,其作用机制与调节角质形成细胞功能以及减少细胞因子和S100a7表达有关。

This study aimed to investigate the therapeutic effect and possible mechanism of carboxyamidotriazole(CAI)on imiquimod(IMQ)-induced psoriasis-like mice model and M5(IL^(-1)α,IL^(-1)7A,IL-22,TNF-αand oncostatin M)-induced keratinocytes model of psoriasis.The severity of psoriasis-like skin lesion in mice was evaluated by psoriasis area and severity index(PASI)score.The histopathological changes of skin were examined by hematoxylin-eosin staining and Baker score was calculated.The levels of pro-inflammatory cytokines in skin were measured by enzyme-linked immunosorbent assay.Transcriptome sequencing technique was used to analyze differentially expressed genes(DEGs)and real-time quantitative PCR(qPCR)was used to detect mRNA expressions.The animal experiments conducted in this study were approved by the Institutional Animal Care Use&Welfare Committee of Institute of Basic Medical Sciences,Chinese Academy of Medical Science(grant No.ACUC-A02-2022-115).The results showed that CAI significantly improved the severity of psoriasis-like lesion,reduced PASI score,attenuated pathological changes,decreased Baker score and inhibited the levels of IL^(-1)β,IL-6,IL^(-1)7A,IL-23 and TNF-αin skin of IMQ-induced mice.Transcriptome sequencing analysis revealed that regulating keratinocytes and their mediated keratinization might be involved in the mechanism of CAI.Further qPCR study validated that CAI down-regulated the mRNA expression of DEG S100a7.Moreover,the keratinocyte model of psoriasis was established by stimulating HaCaT cells by M5.It was shown that CAI decreased the mRNA expression levels of S100a7,Il1β,Il6,Il17,Il23 and Ccl20,which were up-regulated by M5 stimulation.In conclusion,CAI might have a good therapeutic efficacy on psoriasis,and its mechanism was related to regulate the function of keratinocytes and downregulate cytokines and S100a7.