摘要
[目的]探究除湿胃苓汤(Chushi Weiling Decoction,CSWLD)对特异性皮炎(atopic dermatitis,AD)小鼠的皮损改善效果,以及对组织细胞外信号调节激酶1/2(extracellular signal-regulated kinase1/2,ERK1/2)、紧密连接蛋白1(Claudin 1)表达的影响。[方法]采用二硝基氟苯(dinitro fluoro benzene,DNFB)诱导建立小鼠AD模型,随机分为模型组(0.9%氯化钠溶液)和低、中、高CSWLD组(5、10、20 mL·kg^(-1)),另设置空白组(0.9%氯化钠溶液),各组均以相应药物灌胃1周。给药结束后,评价皮损改善情况,并行炎症评分。酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)法检测血清白细胞介素-33(interleukin-33,IL-33)、IL-6、IL-4、CC类趋化因子配体-5(C-C motif chemokine ligand-5,CCL-5)和CCL-2含量。取皮损部位组织,以免疫组化染色测定IL-33、肿瘤抑制素2(suppression of tumorigenicity 2,ST2)、Claudin 1含量;以定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)和免疫印迹检测ERK1/2、Claudin 1的mRNA和蛋白表达。[结果]与空白组比较,模型组小鼠皮肤红肿、干燥、结痂明显,炎症评分增加,血清IL-33、IL-6、IL-4、CCL-5、CCL-2含量升高,组织IL-33、ST2含量增加,Claudin 1 mRNA表达降低,ERK1/2 mRNA表达升高,Claudin 1蛋白表达降低,ERK1/2磷酸化产物(phosphorylated-ERK1/2,p-ERK1/2)与ERK1/2蛋白比值增加(均P<0.01)。在给予不同剂量CSWLD后,小鼠皮损部位红肿消退,结痂脱落,症状减轻。与模型组比较,中、高CSWLD组小鼠炎症评分降低,血清IL-33、IL-6、IL-4、CCL-5、CCL-2含量下降,组织IL-33、ST2含量降低,Claudin 1 mRNA表达升高,ERK1/2 mRNA表达降低,Claudin 1蛋白表达升高,p-ERK1/2与ERK1/2蛋白比值降低(P<0.05,P<0.01);而低CSWLD组小鼠的各项指标差异无统计学意义(P>0.05)。[结论]CSWLD可下调炎性介质,并能抑制组织ERK1/2表达、促进组织Claudin 1表达,缓解AD小鼠皮损症状。
[Objective]To explore treatment effect of Chushi Weiling Decoction(CSWLD)on skin tissue lesion of atopic dermatitis(AD)mice,and its impact on expression of Claudin 1 and extracellular signal-regulated kinase1/2(ERK1/2)in tissue.[Methods]Mice were induced by dinitro fluoro benzene to establish AD model,then randomly divided into model group(0.9%sodium chloride solution),low,medium,high CSWLD groups(5,10,20 mL·kg~(-1)),and set blank group(0.9%sodium chloride solution),all groups took intragastrical administration for 1 week.After administration,the improvement of skin lesion and inflammation scores were evaluated.Enzyme-linked immunosorbent assay(ELISA)assay was used to detect contents of interleukin-33(IL-33),IL-6,IL-4,C-C motif chemokine ligand-5(CCL-5)and CCL-2.Collecting tissue of skin lesion,immunohistochemical staining was used to detect the content of IL-33,suppression of tumorigenicity 2(ST2)and Claudin 1,quantitative polymerase chain reaction(qPCR)and Western blot were used to detect the mRNA and protein expression of ERK1/2 and Claudin 1.[Results]Compared with blank group,model group mice skin was red and swollen,dry and scabbed severely,inflammation score was increased,contents of serum IL-33,IL-6,IL-4,CCL-5 and CCL-2 were increased,contents of IL-33 and ST2 in tissue were increased,Claudin 1 mRNA expression were decreased,ERK1/2 mRNA expression was increased,Claudin 1 protein expression was decreased,phosphorylated-ERK1/2(p-ERK1/2)and ERK1/2 ratio was increased(P<0.01).After the administration with different doses of CSWLD,the redness and swelling of mice skin were degraded,scab separated and symptoms relived.Compared with model group,inflammation scores of mice in medium and high CSWLD groups were decreased,contents of serum IL-33,IL-6,IL-4,CCL-5 and CCL-2 were decreased;in tissue,contents of IL-33 and ST2 were decreased,Claudin 1 mRNA expression was increased,ERK1/2 mRNA expression were decreased,Claudin 1 protein expression were increased,p-ERK1/2 and ERK1/2 ratio were reduced(P<0.05,P<0.01);while there's no statistical difference of results in low CSWLD group(P>0.05).[Conclusion]CSWLD reduced inflammatory factors,inhibited p-ERK1/2/ERK1/2 protein expression and promoted Claudin 1 protein expression in tissue to relieve skin lesion symptoms in AD mice.
作者
徐舰
陈琳瑶
XU Jian;CHEN Linyao(Hangzhou Third People's Hospital,Hangzhou(310009),China)
出处
《浙江中医药大学学报》
CAS
2024年第5期541-547,共7页
Journal of Zhejiang Chinese Medical University
基金
浙江省中医药科学研究基金项目(2021ZA114)。
