标题 | 特应性皮炎小鼠模型中白细胞介素-21、CC趋化因子配体20的表达分析 |
范文 | 王娟 郑爱义 鲍海平
[摘要]目的 测定白细胞介素-21(IL-21)和CC趋化因子配体20(CCL20)在2,4-二硝基氯苯(DNCB)诱导的特应性皮炎小鼠模型中的表达情况,探讨二者在特应性皮炎发病中的作用。方法 将12只BALB/c雌性小鼠,随机分为模型组和对照组,每组6只,使用DNCB构建小鼠特应性皮炎模型。造模成功后采用酶联免疫吸附法检测小鼠血清中IL-21和CCL20的水平。结果 特应性皮炎模型组小鼠血清中IL-21和CCL20的表达水平均高于对照组,差异有统计学意义(P<0.05),并且二者表达成正相关(r=0.59,P<0.05)。结论 IL-21和CCL20在特应性皮炎小鼠模型中表达升高,且成正相关,可能协同参与了特应性皮炎的发生发展。 [关键词]特应性皮炎;白细胞介素-21;CC趋化因子配体20;小鼠模型 [中图分类号] R751? ? ? ? ? [文献标识码] A? ? ? ? ? [文章编号] 1674-4721(2020)8(b)-0014-03 [Abstract] Objective To detect the expression of interleukin-21 (IL-21) and CC chemokine ligand 20 (CCL20) in 2,4-dinitrochlorobenzene (DNCB) induced mice models with atopic dermatitis and explore their roles in the pathogenesis of atopic dermatitis. Methods Twelve female BALB/c mice were randomly divided into the model group and control group(6 in each). Mice models with atopic dermatitis were established by DNCB. After successful modeling, the serum levels of IL-21 and CCL20 were detected by enzyme-linked immunosorbent assay(ELISA). Results The serum levels of IL-21 and CCL20 in the model group with atopic dermatitis were significantly higher than those in the control group (P<0.05). There was a positive correlation between the serum levels of IL-21 and CCL20 in the model group(r=0.59, P<0.05).Conclusions The expression of IL-21 and CCL20 in the mouse model of atopic dermatitis is increased and positively correlated, which may be synergistically involved in the occurrence and development of atopic dermatitis. [Key words] Atopic dermatitis; Interleukin-21; CC chemokine ligand 20; Mouse model 特應性皮炎(AD)是由多种免疫、炎性细胞和细胞因子参与的慢性炎症性皮肤病,皮损顽固难愈,病因和发病机制较为复杂,与遗传、免疫异常、皮肤屏障受损、生存环境和感染等因素有关[1]。不同亚型CD4+T细胞介导的免疫失衡是AD发病的重要环节[2-3]。白细胞介素-21(IL-21)是IL-2家族中重要的多效能细胞因子,可调节淋巴细胞和自然杀伤(NK)细胞的增殖分化,广泛参与体内免疫应答和炎性反应。CC趋化因子配体-20(CCL20)是CC亚族的趋化因子,可趋化表达CC趋化因子受体(CCR)6的淋巴细胞和未成熟树突状细胞向抗原局部定向迁移,与多种自身免疫性疾病有关。本研究通过测定AD小鼠模型血清中IL-21和CCL20的表达变化,探讨二者在AD发病中的作用。 1材料与方法 1.1实验动物与主要试剂 无特定病原体动物(SPF)级7~8周龄健康BALB/c雌性小鼠12只,动物合格证号SCXK(京)2015-0006,由山西大同大学动物实验中心提供。2,4-二硝基氯苯(DNCB)购自美国Sigma公司,小鼠IL-21、CCL20酶联免疫吸附法(ELISA)检测试剂盒购自武汉伊莱瑞特生物科技有限公司。本研究已通过本院医学伦理委员会的审核批准。 1.2方法 1.2.1实验动物造模? 12只BALB/c小鼠清洁级实验室饲养1周,适应环境后开始实验,随机分为模型组和对照组,各6只。参考文献[4-5]构建AD小鼠模型,实验前剃除小鼠背部毛发,面积约3 cm×2 cm。模型组小鼠第1~3天背部剃毛区外涂100 μl 1%DNCB溶液(采用丙酮∶橄榄油=1∶3作为基质配制),第4天继续外涂100 μl 0.1%DNCB溶液,改为每间隔1 d外涂1次,直到第28天涂抹后造模完成。造模后小鼠背部皮肤增厚,可见明显红斑、丘疹、丘疱疹、渗出、堆积大量痂屑。对照组小鼠背部剃毛区仅外涂基质,涂抹剂量、间隔时间与模型组相同。 1.2.2皮肤组织病理检查? 造模后,无菌条件下两组小鼠背部剃毛区皮肤组织取材,10%甲醛溶液固定、脱水、透明、石蜡包埋、切片染色、封片,镜下观察两组小鼠皮肤组织病理改变。 [4]王俊霞,杨子微,车雅敏,等.薏苡仁提取物对BALB/c小鼠特应性皮炎模型的疗效观察及机制探讨[J].中华皮肤科杂志,2018,51(8):609-613. [5]鲍春梅,孙广臣.黑骨藤皂苷对特应性皮炎小鼠的治疗作用及对Th1型免疫反应的影响[J].山东医药,2017,57(13):32-35. [6]刘巍,田文单,Sally A,等.IL-21在Th17细胞炎性反应中的作用[J].国际免疫学杂志,2011,34(4):274-276. [7]Vogelzang A,McGuire HM,Yu D,et al.A fundamental role for interleukin-21 in the generation of T follicular helper cells[J].Immunity,2008,29(1):127-137. [8]杨正生,彭振辉,李晓莉,等.姜黄素对 IL-17 诱导的人表皮角质形成细胞株 CCL20 表达的影响[J].中国皮肤性病学杂志,2011,25(5):346-348. [9]蒋燕,郭顺,邹悦,等.麻黄碱对IL-17诱导的人永生化角质形成细胞HaCaT分泌CCL20的影响[J].山东医药,2017, 57(10):24-27. [10]Caruso R,Fina D,Peluso I,et al.A functional role for interleukin-21 inpromoting the synthesis of the T cell chemoattractant,MIP-3alpha,by gut epithelial cells[J].Gastroenterology,2007,132(1):166-175. [11]Orciani M,Campanati A,Caffarini M,et al.T helper (Th)1,Th17 and Th2 imbalance in mesenchymal stem cells of adult patients with atopic dermatitis: at the origin of the problem[J].Br J Dermatol,2017,176(6):1569-1576. [12]蔣有让,刁庆春,史丙俊,等.特应性皮炎患者血清中Th17和Treg相关细胞因子的检测[J].临床皮肤科杂志,2016,45(6):411-413. [13]Szabo,K,Gaspar K,Dajnoki Z,et all.Expansion of circulating follicular T helper cells associates with disease severity in childhood atopic dermatitis[J].Immunol Lett,2017,189(9):101-108. [14]Jin H,Oyoshi MK,Le Y,et al.IL-21R is essential for epicutaneous sensitization and allergic skin inflammation in humans and mice[J].J Clin Invest,2009,119(1):47-60. [15]Mizutani H,Tamagawa-Mineoka R,Nakamura N,et al.Serum IL-21 levels are elevated in atopic dermatitis patients with acute skin lesions[J].Allergol Int,2017,66(3):440-444. [16]邱会芬,王雯,王忠永,等.NB-UVB对特应性皮炎患者外周血IL-23/IL-17轴表达的影响[J].中国皮肤性病学杂志,2016,30(10):1010-1013. [17]Lin SC,Chuang YH,Yang YH,et al.Decrease in interleukin-21 in children suffering with severe atopic dermatitis[J].Pediatr Allergy Immunol,2011,22(8):869-875. [18]Machura E,Rusek-Zychma M,Jachimowicz M,et al.Serum TARC and CTACK concentrations in children with atopic dermatitis,allergic asthma,and urticaria[J].Pediatr Allergy Immunol,2012,23(3):278-284. [19]Miyano K,Matsushita S,Tsuchida T,et al.Inhibitory effect of a histamine 4 receptor? antagonist on CCL17 and CCL22 production by monocyte-derived Langerhans cells in patients with atopic dermatitis[J].J Dermatol,2016,43(9):1024-1029. [20]Park CW,Lee BH,Han HJ,et al.Tacrolimus decreases the expression of eotaxin,CCR3,RANTES and interleukin-5 in atopic dermatitis[J].Br J Dermatol,2005,152(6):1173-1181. (收稿日期:2019-12-13) |
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