穿心莲内酯调节cGAS-STING信号通路对银屑病小鼠的治疗作用  

Study on the therapeutic effect of andrographolide on psoriasis mouse model by regulating cGAS-STING signal pathway

作　　者：何亚男 蔡翔[1] 邱百怡[1] 孙邦梅[1] 李伶华 HE Yanan;CAI Xiang;QIU Baiyi;SUN Bangmei;LI Linghua(Department of Dermatology,Wuhan Traditional Chinese Medicine Hospital,Wuhan 430050,China)

机构地区：[1]武汉市中医医院皮肤科,430050

出　　处：《天津医药》2024年第4期379-386,共8页Tianjin Medical Journal

基　　金：武汉市中医药科研项目(WZ22C75,WZ22C72)。

摘　　要：目的 探究穿心莲内酯(Andro)调节GMP-AMP合酶-干扰素基因刺激物(cGAS-STING)信号通路对银屑病小鼠的治疗作用及机制。方法 90只BALB/c小鼠分为对照组(Control组),模型组(Model组),穿心莲内酯低、中、高剂量组(Andro-L、M、H组,10、30、50 mg·kg^(-1)·d^(-1) Andro)和穿心莲内酯高剂量+STING激活剂DMXAA组(Andro-H+DMXAA组,50 mg·kg^(-1)·d^(-1) Andro+5 mg·kg^(-1)·d^(-1) DMXAA)。除Control组外,其余组小鼠背部施用咪喹莫特(IMQ)建立银屑病小鼠模型。观察小鼠银屑病面积并进行严重程度指数(PASI)评分,酶联免疫吸附试验(ELISA)检测血清白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α、IL-1β、IL-23、IL-17A、干扰素(IFN)-γ和IFN-β水平,苏木素-伊红(HE)染色和甲苯胺蓝染色测定表皮厚度和肥大细胞数,免疫荧光染色检测Ki-67、CD4阳性细胞表达率,RT-q PCR检测c GAS和STING m RNA表达水平,免疫印迹实验检测c GAS、STING、干扰素调节因子3(IRF3)、p-IRF3蛋白水平。结果 与Control组相比,Model组小鼠背部皮肤出现严重的红斑、鳞屑,有大量的炎性细胞浸润;抓挠次数、PASI评分、表皮厚度、肥大细胞数量、IL-6、IL-1β、TNF-α、IL-23、IL-17A、IFN-γ和IFN-β水平、Ki-67和CD4阳性表达率、c GAS和STING m RNA和蛋白表达水平及IRF3磷酸化水平升高(P<0.05)。与Model组相比,Andro-L组、Andro-M组和Andro-H组小鼠皮肤红斑、鳞屑、炎性细胞浸润现象减轻;抓挠次数、PASI评分、表皮厚度、肥大细胞数量、IL-6、IL-1β、TNF-α、IL-23、IL-17A、IFN-γ和IFN-β水平、Ki-67和CD4阳性表达率、cGAS和STING mRNA和蛋白表达水平及IRF3磷酸化水平呈剂量依赖性降低(P<0.05);STING激活剂DMXAA逆转了穿心莲内酯对银屑病小鼠的保护作用(P<0.05)。结论 穿心莲内酯可通过抑制cGAS-STING信号通路减轻炎症反应,改善小鼠银屑病症状。Objective To explore the therapeutic mechanism of andrographolide(Andro)on psoriasis mice by regulating GMP-AMP synthase-stimulator of interferon genes signal pathway(cGAS STING signal pathway).Methods Ninety BALB/c mice were divided into the control group,the model group,the Andro low dose group,the Andro medium dose group,the Andro high dose groups(Andro-L,M,H groups,10,30 and 50 mg·kg^(-1)·d^(-1) Andro)and the high dose Andro+STING activator DMXAA group(Andro-H+DMXAA group,50 mg·kg^(-1)·d^(-1) Andro+5 mg·kg^(-1)·d^(-1) DMXAA).Except for the control group,mice of the other groups were administered imiquimod(IMQ)on back to establish the psoriasis mouse model.The psoriasis area of mice was observed,and severity index(PASI)scoring was performed.Serum levels of interleukin-6(IL 6),tumor necrosis factorα(TNF-α),interleukin(IL)^(-1)β,IL-23,IL^(-1)7A,interferon(IFN)-γand IFN-βwere detected by enzyme-linked immunosorbent assay(ELISA).Hematoxylin eosin(HE)staining and toluidine blue staining were used to detect the epidermal thickness and mast cell count.Immunofluorescence staining was applied to detect the expression rates of Ki-67 and CD4 positive cells.RT-qPCR was applied to detect the expression level of cGAS and STING mRNA,and Western blot assay was applied to detect levels of cGAS,STING,interferon regulatory factor-3(IRF3)and p-IRF3 proteins.Results Compared with the control group,the back skin of mice in the model group showed severe erythema,scales and a large number of inflammatory cell infiltration.The scratch times,PASI score,epidermal thickness,number of mast cells,levels of IL-6,IL^(-1)β,TNF-α,IL-23,IL^(-1)7A,IFN-γand IFN-β,the expression rates of Ki-67 and CD4 positive cell,the expression level of cGAS and STING mRNA and protein,and IRF3 phosphorylation level were increased obviously(P<0.05).Compared with the model group,skin erythema,scales and inflammatory cell infiltration were reduced in the Andro-L,Andro-M and Andro-H groups.The scratch times,PASI score,epidermal thickness,number of mas

关 键 词：穿心莲内酯 银屑病 炎症 GMP-AMP合酶-干扰素基因刺激物信号通路 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]