基于Nrf2/HO-1/NF-κB信号通路探讨马齿苋外用对变应性接触性皮炎小鼠的作用机制  

Exploration on Mechanism of Topical Treatment of Allergic Contact Dermatitis in Mice with Portulacae Herba Based on Nrf2/HO-1/NF-κB Signaling Pathway

作　　者：王小雪 樊官伟[2] 蒲翔 张仲昭 陈霞 唐英 吴娜娜 骆江利 孔祥艳 WANG Xiaoxue;FAN Guanwei;PU Xiang;ZHANG Zhongzhao;CHEN Xia;TANG Ying;WU Nana;LUO Jiangli;KONG Xiangyan(Guizhou University of Traditional Chinese Medicine(TCM),Guiyang 550025,China;First Teaching Hospital of Tianjin University of TCM,Tianjin 300193,China;The First Affiliated Hospital of Guizhou University of TCM,Guiyang 550001,China)

机构地区：[1]贵州中医药大学,贵阳550025 [2]天津中医药大学第一附属医院,天津300193 [3]贵州中医药大学第一附属医院,贵阳550001

出　　处：《中国实验方剂学杂志》2025年第3期115-123,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：贵州中医药大学国家与省级科技创新人才团队培育项目(贵中医TD[2022]002号);贾敏全国名老中医药专家传承工作室项目(国中医药人教函[2022]75号)。

摘　　要：目的:基于核因子E_(2)相关因子2(Nrf2)/血红素加氧酶-1(HO-1)/核转录因子-κB(NF-κB)信号通路探讨马齿苋外用对变应性接触性皮炎(ACD)小鼠的保护作用机制。方法:将70只6周龄SPF级雌性昆明种小鼠适应性喂养1周,随机分成空白组、模型组、复方醋酸地塞米松乳膏组(2.075×10^(-2) g·g^(-1))、空白基质乳膏组、马齿苋低剂量乳膏组(0.1 g·g^(-1))、马齿苋高剂量乳膏组(0.2 g·g^(-1))、马齿苋+抑制剂组(0.2 g·g^(-1)+30 mg·kg^(-1)ML385),每组10只。于实验前1 d对小鼠进行颈背部剃毛处理,除正常组外其余各组小鼠采用2,4-二硝基氯苯(DNCB)建立ACD模型,经分组给药处理后,观察小鼠皮肤皮损评分、苏木素-伊红(HE)染色皮肤组织病理学变化,酶联免疫吸附测定法(ELISA)检测小鼠血清中白细胞介素-6(IL-6)、IL-1β、活性氧(ROS)、超氧化物歧化酶(SOD)活性、丙二醛(MDA)水平,免疫组化(IHC)、蛋白免疫印迹法(Western blot)、实时荧光定量聚合酶链式反应(Real-time PCR)检测小鼠皮肤组织Nrf2/HO-1/NF-κB信号通路相关蛋白表达。结果:与空白组比较,模型组小鼠皮肤皮损评分升高(P<0.01),皮肤组织出现严重的病理损伤,小鼠血清中IL-1β、IL-6、ROS、MDA含量上升(P<0.01),SOD活性下降(P<0.01),皮肤组织中Nrf2、HO-1、核转录因子-κB抑制因子α(IκBα)mRNA及蛋白表达水平上调(P<0.01),磷酸化(p)-IκBα、p-NF-κB p65蛋白及NF-κB p65 mRNA表达水平下调(P<0.01);与模型组及空白基质乳膏组比较,经马齿苋治疗后皮肤皮损评分下降(P<0.01),皮肤组织病理损伤减轻,血清中IL-1β、IL-6、ROS、MDA含量降低(P<0.01),SOD活性上升(P<0.01),皮肤组织中Nrf2、HO-1、IκBαmRNA及蛋白表达水平下调(P<0.05,P<0.01),p-IκBα、p-NF-κB p65蛋白及NF-κB p65 mRNA表达水平上调(P<0.05,P<0.01);与马齿苋+抑制剂组比较,马齿苋高剂量乳膏组皮肤皮损评分降低(P<0.01),皮肤组织病理损伤减轻,小鼠血清中IL-1�Objective:To investigate the mechanism of topical treatment of allergic contact dermatitis(ACD)mice with Portulacae Herba based on the nuclear factor E_(2)-related factor 2(Nrf2)/heme oxygenase-1(HO-1)/nuclear factor-κB(NF-κB)signaling pathway.Methods:A total of 706-week-old specific pathogen free(SPF)female Kunming mice were adaptively fed for 1 week and randomly divided into blank group,model group,compound dexamethasone acetate cream group(2.075×10^(-2) g·g^(-1)),blank matrix cream group,low-dose Portulacae Herba cream group(0.1 g·g^(-1)),high-dose Portulacae Herba cream group(0.2 g·g^(-1)),and Portulacae Herba+inhibitor group(0.2 g·g^(-1)+30 mg·kg^(-1)ML385),with 10 mice in each group.One day before the experiment,the mice were shaved on the neck and back.Except for the blank group,the mice in the other groups were treated with 2,4-dinitrochlorobenzene(DNCB)to establish an ACD model.After respective administration,the skin lesion of the mice was scored,and the histopathological changes of the skin were stained with hematoxylin-eosin(HE).Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-6(IL-6),interleukin-1β(IL-1β),reactive oxygen species(ROS),superoxide dismutase(SOD)activity,and malondialdehyde(MDA)in serum of mice.The expression of Nrf2/HO-1/NF-κB signaling pathway-related proteins in mouse skin tissue was detected by immunohistochemistry(IHC),Western blot,and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Results:Compared with the blank group,the mice in the model group had an increased skin lesion score(P<0.01),severe pathological damage to skin tissue,increased content of IL-1β,IL-6,ROS,and MDA in their serum(P<0.01),and decreased content of SOD(P<0.01).In addition,the mRNA and protein expression levels of Nrf2,HO-1,and nuclear factor-κB inhibitorα(IκBα)in skin tissue were up-regulated(P<0.01),while the protein expression levels of phosphorylated(p)-IκBαand p-NF-κB p65 and the mRNA expression of NF-κB p65 were down-regu

关 键 词：马齿苋 外用治疗 核因子E_(2)相关因子2(Nrf2)/血红素加氧酶-1(HO-1)/核转录因子-κB(NF-κB) 变应性接触性皮炎 作用机制 

分 类 号：R275[医药卫生—中医皮科] R285[医药卫生—中医学] R751