组蛋白脱乙酰酶3抑制通过调控Th17促进小鼠银屑病的发展  

Inhibition of HDAC3 Promotes Psoriasis Development in Mice Through Regulating Th17

作　　者：许帆 张辛瑞 夏阳晨 李文婷 陈昊 秦安琪 张爱红 朱依然 田枫 郑全辉 XU Fan;ZHANF Xin-Rui;XIA Yang-Chen;LI Wen-Ting;CHEN Hao;QIN An-Qi;ZHANG Ai-Hong;ZHU Yi-Ran;TIAN Feng;ZHENG Quan-Hui(Hebei Key Laboratory for Chronic Diseases,School of Basic Medical Sciences,North China University of Science and Technology,Tangshan 063210,China;Department of Laboratory Animal Science,Health Science Center,Peking University,Beijing 100083,China;Intensive Care Unit,the Affiliated Hospital of North China University of Science and Technology,Tangshan 063000,China)

机构地区：[1]华北理工大学基础医学院,河北省慢性疾病基础医学重点实验室,唐山063210 [2]北京大学医学部实验科学部,北京100083 [3]华北理工大学附属医院重症医学科,唐山063000

出　　处：《生物化学与生物物理进展》2025年第4期1008-1017,共10页Progress In Biochemistry and Biophysics

基　　金：河北省卫健委(20190105);国家自然科学基金(81373111)资助项目。

摘　　要：目的 探讨组蛋白脱乙酰酶3 (histone deacetylase 3,HDAC3)对小鼠银屑病样炎症发生发展的影响及相关免疫机制。方法 选取6~8周龄健康C57BL/6小鼠,将小鼠随机分为对照组(Control),银屑病模型组(IMQ),HDAC3抑制剂RGFP966处理的银屑病模型组(IMQ+RGFP966),提前1 d对小鼠进行剃毛处理。待稳定1 d后,Control组涂抹等量的凡士林,IMQ组背部涂抹咪喹莫特(imiquimod,IMQ,62.5 mg/d),建立小鼠银屑病样炎症模型;IMQ+RGFP966组在银屑病模型基础上以高剂量HDAC3选择性抑制剂RGFP966 (30 mg/kg)进行干预处理。各组持续处理5 d,观察记录背部皮肤银屑病样炎症症状(鳞屑、红斑、皮肤厚度)、体重变化及精神状态,并拍照存档。在小鼠解剖后,采用苏木精-伊红(hematoxylin-eosin,HE)染色检测RGFP966对银屑病模型小鼠皮肤组织层次结构的影响,并测量皮肤厚度。通过实时荧光定量PCR (reverse transcription real-time quantitative polymerase chain reaction,RT-qPCR)和蛋白质印迹法(Western blot,WB),分别检测皮肤组织中HDAC3的m RNA和蛋白质表达水平。采用流式细胞术检测各组小鼠外周血和淋巴结中性粒细胞、外周血CD4^(+)T淋巴细胞和CD8^(+)T淋巴细胞及外周血CD4^(+)T淋巴细胞的白介素-17A (interleukin-17A,IL-17A)分泌水平,检测脾脏CD4^(+)T淋巴细胞HDAC3、CC基序趋化因子受体(CC motif chemokine receptor,CCR) 6、CCR8表达及IL-17A分泌水平。采用免疫组化检测皮肤HDAC3、IL-17A、白介素-10 (interleukin-10,IL-10)水平。结果 与Control组相比,IMQ组小鼠展示出明显的银屑病样炎症,出现红斑、鳞屑及皮肤褶皱。RGFP966加重了银屑病样炎症症状,皮肤角化增多。银屑病面积与严重性指数(psoriasis area and severity index,PASI)皮肤症状评分,IMQ组高于Control,IMQ+RGFP966组高于IMQ组。测量各组皮肤厚度,IMQ+RGFP966>IMQ>Control。Control、IMQ、IMQ+RGFP966组的血液和淋巴结的中性粒细胞依次增多,血液CD4^(+)T淋巴Objective To investigate the influence of histone deacetylase 3(HDAC3)on the occurrence,development of psoriasis-like inflammation in mice,and the relative immune mechanisms.Methods Healthy C57BL/6 mice aged 6-8 weeks were selected and randomly divided into 3 groups:control group(Control),psoriasis model group(IMQ),and HDAC3 inhibitor RGFP966-treated psoriasis model group(IMQ+RGFP966).One day prior to the experiment,the back hair of the mice was shaved.After a one-day stabilization period,the mice in Control group was treated with an equal amount of vaseline,while the mice in IMQ group was treated with imiquimod(62.5 mg/d)applied topically on the back to establish a psoriasis-like inflammation model.The mice in IMQ+RGFP966 group received intervention with a high dose of the HDAC3-selective inhibitor RGFP966(30 mg/kg)based on the psoriasis-like model.All groups were treated continuously for 5 d,during which psoriasis-like inflammation symptoms(scaling,erythema,skin thickness),body weight,and mental status were observed and recorded,with photographs taken for documentation.After euthanasia,hematoxylin-eosin(HE)staining was used to assess the effect of RGFP966 on the skin tissue structure of the mice,and skin thickness was measured.The mRNA and protein expression levels of HDAC3 in skin tissues were detected using reverse transcription real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot(WB),respectively.Flow cytometry was employed to analyze neutrophils in peripheral blood and lymph nodes,CD4^(+)T lymphocytes,CD8^(+)T lymphocytes in peripheral blood,and IL-17A secretion by peripheral blood CD4^(+)T lymphocytes.Additionally,spleen CD4^(+)T lymphocyte expression of HDAC3,CCR6,CCR8,and IL-17A secretion levels were analyzed.Immunohistochemistry was used to detect the localization and expression levels of HDAC3,IL-17A,and IL-10 in skin tissues.Results Compared with the Control group,the IMQ group exhibited significant psoriasis-like inflammation,characterized by erythema,scaling,and skin wrinkling.C

关 键 词：组蛋白去乙酰化酶3 IL-17 银屑病 CCR8 

分 类 号：R758.63[医药卫生—皮肤病学与性病学] R-332[医药卫生—临床医学]