清除Vγ4T细胞在小鼠皮肤紫外线损伤后表皮组织修复中的作用及其机制  

Role and mechanism of Vγ4 T cell depletion in epidermal tissue repair after ultraviolet damage to mouse skin

作　　者：李雅舒 贺伟峰 吕开阳 Li Yashu;He Weifeng;Lyu Kaiyang(Department of Plastic Surgery,Xinhua Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China;Institute of Burn Research,State Key Laboratory of Trauma and Chemical Poisoning,the First Affiliated Hospital of Army Medical University(the Third Military Medical University),Chongqing Key Laboratory for Wound Damage Repair and Regeneration,Chongqing 400038,China)

机构地区：[1]上海交通大学医学院附属新华医院整形外科,上海200092 [2]陆军军医大学(第三军医大学)第一附属医院全军烧伤研究所,创伤与化学中毒全国重点实验室,重庆市创面损伤修复与再生重点实验室,重庆400038

出　　处：《中华烧伤与创面修复杂志》2024年第5期415-424,共10页Chinese Journal of Burns And Wounds

基　　金：国家自然科学基金青年科学基金项目(32000645)。

摘　　要：目的探究清除Vγ4T细胞在小鼠皮肤紫外线损伤后表皮组织修复中的作用及其机制。方法该研究为实验研究。将54只6~8周龄雌性C57BL/6J野生型小鼠按照随机数字表法分为Vγ4T细胞清除组和对照组(每组27只),分别腹腔注射亚美尼亚仓鼠抗小鼠Vγ4T细胞受体(TCR)单克隆抗体200µg、等量同型对照IgG抗体。注射后1周(之后均在此时间点取小鼠),每组取3只小鼠(之后均从这2组另取小鼠),从背部皮肤组织和腋窝、腹股沟淋巴结中分别提取真皮细胞和淋巴结细胞,行流式细胞术检测真皮细胞和淋巴结细胞中Vγ4T细胞比例;每组取5只小鼠,观察背部皮肤情况,之后行苏木精-伊红(HE)染色观察皮肤组织结构并测量表皮组织厚度;每组取5只小鼠,提取表皮细胞后行流式细胞术检测表皮细胞中树突状表皮T细胞(DETC)比例。每组取3只小鼠,分别设为Vγ4T细胞清除+5次紫外线辐射(UVR)组和对照+5次UVR组,每日1次共行5次UVR,每日照射后即刻观察背部皮肤情况;每组取5只小鼠,分别设为Vγ4T细胞清除+1次UVR组和对照+1次UVR组,均行1次UVR后即刻行HE染色后测量表皮组织厚度。每组取3只小鼠,分别设为单纯Vγ4T细胞清除组、单纯对照组;再另取3只小鼠,分别设为Vγ4T细胞清除+1次UVR组和对照+1次UVR组;均同前处理后,采用实时荧光定量反转录PCR法检测表皮组织中胰岛素样生长因子Ⅰ(IGF-Ⅰ)、角质形成细胞生长因子(KGF)、Vγ5TCR、白细胞介素15(IL-15)、IL-1β、IL-23、自然杀伤细胞2族成员D(NKG2D)、组织相容性抗原60(H60)、小鼠UL16结合蛋白样转录子1(Mult1)、维甲酸早期诱导蛋白1(Rae1)的mRNA表达。结果注射后1周,Vγ4T细胞清除组小鼠真皮细胞、淋巴结细胞中Vγ4T细胞比例均明显低于对照组(t值分别为27.99、13.12,P<0.05);Vγ4T细胞清除组和对照组小鼠皮肤大体情况和组织结构无明显区别,表皮组织厚度相近(P>0.05);Vγ4T细胞清除组�Objective To explore the role and mechanism of Vγ4 T cell depletion in epidermal tissue repair after ultraviolet damage to mouse skin.Methods The study was an experimental study.Fifty-four female C57BL/6J wild-type mice aged 6 to 8 weeks were divided into Vγ4 T cell depletion group and control group(27 mice in each group)according to the random number table,and the Armenian hamster anti-mouse Vγ4 T cell receptor(TCR)monoclonal antibody of 200µg and an equal amount of homologous control IgG antibody were intraperitoneally injected,respectively.At one week after injection(the same time point to harvest mice below),dermal cells and lymph node cells were respectively extracted from the back skin tissue,armpit and inguinal lymph nodes of 3 mice in each group(mice in following study were all taken from these 2 groups),and the proportions of Vγ4 T cells in dermal cells and lymph node cells were detected by flow cytometry.Five mice from each group were harvested for observation of skin on the back and skin tissue structure was observed and the epidermal tissue thickness was measured after hematoxylin-eosin(HE)staining.Five mice from each group were harvested for detection of proportion of dendritic epidermal T cells(DETCs)in epidermal cells by flow cytometry after extracted.Three mice were taken from each group and recruited in Vγ4 T cell depletion+5 times ultraviolet irradiation(UVR)group and control+5 times UVR group,respectively,then UVR was administered once per day for 5 times,and the condition of skin on the back was observed immediately after daily irradiation.Five mice were taken from each group and divided into Vγ4 T cell depletion+1 UVR group and control+1 UVR group,respectively.Immediately after one UVR treatment,the epidermal tissue thickness was measured after HE staining.Three mice from each group were selected and recruited in Vγ4 T cell depletion alone group and control alone group,then 3 mice from each group rwere recruited in Vγ4 T cell depletion+1 time UVR group and control+1 time UVR group,re

关 键 词：紫外线 皮肤 表皮 胰岛素样生长因子Ⅰ Vγ4T细胞 树突状表皮T细胞 自然杀伤细胞2族成员D 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]