紫草素调节cGAS/STING信号通路对银屑病细胞模型炎症反应的影响  

Shikonin Inhibits Inflammation of Psoriasis Cell Model by Regulating cGAS/STING Signaling Pathway

作　　者：吕冲 乔现华 高娟娟[2] 田菲[2] 周奎龙[2] 王成成 王杰鹏 LYU Chong;QIAO Xianhua;GAO Juanjuan;TIAN Fei;ZHOU Kuilong;WANG Chengcheng;WANG Jiepeng(Hebei University of Chinese Medicine,Shijiazhuang 050000,China;Xingtai People's Hospital,Xingtai 054000,China)

机构地区：[1]河北中医药大学,石家庄050000 [2]邢台市人民医院,河北邢台054000

出　　处：《中国实验方剂学杂志》2024年第24期114-120,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省自然科学基金项目(H2023423061);邢台市重点研发计划项目(2021ZC120)。

摘　　要：目的:探讨紫草素(SHI)介导环鸟苷酸-腺苷酸合成酶(cGAS)/干扰素基因刺激蛋白(STING)信号通路调控炎症反应防治银屑病(PSO)的作用及机制。方法:以HaCaT细胞为研究对象,将其分为正常组(Control组)、五联因子(M5)组[添加10μg·L^(-1)的白细胞介素(IL)-1α、IL-17、IL-22、肿瘤坏死因子-α(TNF-α)、抑瘤素M(OSM)刺激细胞48 h]、SHI低、中、高剂量组(L-SHI、M-SHI、H-SHI组,在M5诱导的基础上添加0.1、1、10μmol·L^(-1)的SHI)、SHI高剂量+ADU-S100组(SHI+ADU-S100组,在H-SHI组的基础上添加10μmol·L^(-1) STING激活剂ADU-S100)。噻唑蓝(MTT)比色法和平板克隆实验检测SHI对HaCaT细胞增殖的影响;划痕实验检测SHI对HaCaT细胞迁移能力的影响;流式细胞仪检测SHI对HaCaT细胞凋亡的影响;酶联免疫吸附测定法(ELISA)检测HaCaT细胞中IL-1β、IL-6、IL-15、IL-23、γ干扰素(IFN-γ)炎症因子的表达;蛋白免疫印迹法(Western blot)检测HaCaT细胞中cGAS、STING蛋白表达。结果:与Control组比较,M5组HaCaT细胞的存活率、克隆细胞数、划痕愈合率降低,凋亡率、IL-1β、IL-6、IL-15、IL-23、IFN-γ、cGAS、STING蛋白表达显著升高(P<0.01);与M5组比较,L-SHI组、M-SHI组、H-SHI组存活率、克隆细胞数、划痕愈合率升高,凋亡率、IL-1β、IL-6、IL-15、IL-23、IFN-γ、cGAS、STING蛋白表达显著降低(P<0.01);与H-SHI组比较,SHI+ADU-S100组存活率、克隆细胞数、划痕愈合率降低,凋亡率、IL-1β、IL-6、IL-15、IL-23、IFN-γ、cGAS、STING蛋白表达显著升高(P<0.01)。结论:SHI可以抑制PSO细胞模型的炎症反应,其机制可能是通过抑制cGAS/STING信号通路实现的。Objective:To investigate the effect of shikosin(SHI)on psoriasis(PSO)and explore the underlying mechanism via the cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)/stimulator of interferon genes(STING)signaling pathway.Method:HaCaT cells were classified into normal culture(Control),a mixture of five proinflammatory cytokines(M5),low-,medium-,and high-dose SHI(L-SHI,M-SHI,and H-SHI,respectively),and SHI+ADU-S100 groups.The cells in the M5 group were stimulated with 10μg·L^(-1) interleukin(IL)-1α,IL-17,IL-22,tumor necrosis factor(TNF)-α,and oncostatin M(OSM)for 48 h.The cells in the L-SHI,M-SHI,and H-SHI groups were treated with 0.1,1,10μmol·L^(-1)SHI,respectively,on the basis of the treatment in the M5 group.The cells in the SHI+ADU-S100 group were treated with 10μmol·L^(-1) STING activator ADU-S100 on the basis of the treatment in the H-SHI group.The methyl thiazolyl tetrazolium(MTT)assay and colony formation assay were employed to examine the effect of SHI on the proliferation of HaCaT cells.The wound healing assay was employed to examine the effect of SHI on the migration of HaCaT cells.Flow cytometry was employed to detect the effect of SHI on the apoptosis of HaCaT cells.Enzyme-linked immunosorbent assay was employed to measure the levels of IL-1β,IL-6,IL-15,IL-23,and interferon-γ(IFN-γ)in HaCaT cells.Western blot was employed to determine the protein levels of cGAS and STING in HaCaT cells.Result:Compared with Control group,the M5 group showed decreased survival rate,colony formation,and would healing rate of HaCaT cells,increased apoptosis rate,elevated levels of IL-1β,IL-6,IL-15,IL-23,and IFN-γ,and up-regulated protein levels of cGAS and STING(P<0.01).Compared with the M5 group,the L-SHI,M-SHI,and H-SHI groups showed increased survival rate,cell colony formation,and wound healing rate,decreased apoptosis rate,lowered levels of IL-1β,IL-6,IL-15,IL-23,and IFN-γ,and down-regulated protein levels of cGAS and STING(P<0.01).Compared with the H-SHI group,the SHI+ADU-S100 group s

关 键 词：紫草素 环鸟苷酸-腺苷酸合成酶 干扰素基因刺激蛋白 银屑病 炎症反应 

分 类 号：R2-0[医药卫生—中医学] R33R289R24