养血解毒方对慢性银屑病小鼠皮损组织驻留记忆T细胞的影响  

Effects of Yangxue Jiedu Formula on Skin Lesion Tissue-resident Memory T Cells in Mice with Chronic Psoriasis

作　　者：陈岩[1] 刘雨琛 马慧可 赵京霞[1] CHEN Yan;LIU Yuchen;MA Huike;ZHAO Jingxia(Traditional Chinese Medicine Hospital of Beijing Affiliated to Capital Medical University/Beijing Institute of Traditional Chinese Medicine/Beijing Key Laboratory of Clinical Basic Research of Traditional Chinese Medicine for Psoriasis,Beijing China 100010)

机构地区：[1]首都医科大学附属北京中医医院/北京市中医药研究所/银屑病中医临床基础研究北京市重点实验室,北京100010

出　　处：《中医学报》2025年第3期628-634,共7页Acta Chinese Medicine

基　　金：国家自然科学基金项目(82074434)。

摘　　要：目的:探讨养血解毒方对咪喹莫特诱导的慢性银屑病小鼠皮损组织驻留记忆T细胞(tissue resident memory T cell,TRM)浸润的影响。方法:从24只Balb/c小鼠中随机选取6只作为正常组,其余小鼠在背部皮肤涂抹5%咪喹莫特乳膏,每日42 mg,连续5 d,制备慢性银屑病模型。将造模成功的小鼠随机分为模型组、养血解毒方组(YXJD,24.57 g·kg^(-1))和甲氨蝶呤组(methotrexate,MTX,1 mg·kg^(-1)),每组6只。从第6天开始,除正常组外,其余小鼠均隔天同量涂抹咪喹莫特乳膏至第14天。各组小鼠以纯净水或相应药物灌胃,持续7 d。采用PASI评分判断小鼠皮损程度;HE染色观察小鼠皮损组织病理形态;ELISA法检测小鼠血清白细胞介素(interleukin,IL)-17、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、IL-22含量;免疫荧光法检测小鼠皮损组织Ki67表达水平和CD8+CD103+T细胞数目;荧光定量PCR法检测小鼠皮损组织中IL-17A、IL-17F、IL-22、Fabp4、Fabp5、CD36基因表达水平。结果:实验第6天与第14天,与正常组比较,模型组小鼠的背部皮损红斑、鳞屑、增厚评分及总PASI评分升高(P<0.01);与模型组比较,YXJD组和MTX组小鼠的背部皮损红斑、鳞屑、增厚评分及总PASI评分降低(P<0.01)。HE染色显示,正常组小鼠的皮肤表皮层较薄,真皮层中淋巴细胞浸润较少。与正常组比较,模型组小鼠的表皮层增厚(P<0.01),真皮中大量淋巴细胞浸润。与模型组比较,YXJD组和MTX组小鼠的表皮厚度降低(P<0.01),真皮中淋巴细胞浸润减少。与正常组比较,模型组小鼠皮损组织中Ki67蛋白表达水平和CD8+CD103+T细胞数目增加(P<0.01)。与模型组比较,YXJD组和MTX组小鼠皮损组织中Ki67蛋白表达水平和CD8+CD103+T细胞数目减少(P<0.01)。与正常组比较,模型组小鼠血清IL-17、TNF-α含量升高(P<0.01);与模型组比较,YXJD组和MTX组小鼠血清IL-17、TNF-α含量降低(P<0.01)。与正常组比较,模型组小鼠皮损组织IL-Objective:To investigate the effect of Yangxue Jiedu Formula on imiquimod-induced infiltration of tissue resident memory T cells(TRM)in skin lesions of chronic psoriasis mice.Methods:A chronic psoriasis model was prepared by randomly selecting 6 out of 24 Balb/c mice as the normal group,and the rest of the mice were coated with 5%imiquimod cream,42 mg per day for 5 d on the back skin.The successfully modeled mice were randomly divided into model group,Yangxue Jiedu Formula group(YXJD,24.57 g·kg^(-1))and the methotrexate group(methotrexate,MTX,1 mg·kg^(-1)),with 6 mice in each group.Starting from day 6,imiquimod cream was applied in the same amount every other day until day 14 in all mice except the normal group.The mice in each group were gavaged with pure water or corresponding drugs for 7 d.The PASI score was used to determine the degree of skin lesions in mice;HE staining was used to observe the histopathological morphology of the skin lesions in mice;ELISA was used to detect the levels of interleukin(IL)-17,tumor necrosis factor-α(TNF-α)and IL-22;immunofluorescence was performed to determine the levels of IL-22 and TNF-αin the blood serum of mice,IL-22;Ki67 expression level and CD8+CD103+T-cell number were detected by immunofluorescence;IL-17A,IL-17F,IL-22,Fabp4,Fabp5,CD36 gene expression level was detected by fluorescence quantitative PCR in mouse skin lesion tissues.Results:On day 2 and day 14 of the experiment,the erythema,scaling,thickening scores and total PASI scores of the back skin lesions of the mice in the model group were elevated compared with those of the normal group(P<0.01);the erythema,scaling,thickening scores and total PASI scores of the back skin lesions of the mice in the YXJD group and the MTX group decreased compared with those of the model group(P<0.01).HE staining showed that the skin of the normal group mice epidermis was thinner,and there was less lymphocyte infiltration in the dermis.Compared with the normal group,mice in the model group had a thickened epidermal layer(P<0.01)a

关 键 词：养血解毒方 慢性银屑病 组织驻留记忆T细胞 免疫浸润 

分 类 号：R285.5[医药卫生—中药学]