1.上海中医药大学中药研究所,国家中医药管理局中药新资源与品质评价重点研究室,上海市复方中药重点实验室(上海 201203)
2.上海交通大学医学院附属新华医院崇明分院(上海 202150)
韩欣妍,女,博士研究生,主要从事中药药理与免疫方面研究工作
杨柳,博士后;E-mail: yangliu996633@126.com
吴晓俊,研究员,博士研究生导师;E-mail: xiaojunwu320@126.com
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韩欣妍,陈姿羽,袁金凤,等.黄花蒿茶提取物抑制Th2细胞相关炎症因子表达减轻DNCB诱导BALB/c小鼠特应性皮炎[J].上海中医药杂志,2022,56(12):73-78.
HAN Xinyan,CHEN Ziyu,YUAN Jinfeng,et al.Artemisia annua tea extract attenuating DNCB⁃induced atopic dermatitis by restraining Th2 cell mediated inflammatory responses in BALB/c mice[J].Shanghai Journal of Traditional Chinese Medicine,2022,56(12):73-78.
韩欣妍,陈姿羽,袁金凤,等.黄花蒿茶提取物抑制Th2细胞相关炎症因子表达减轻DNCB诱导BALB/c小鼠特应性皮炎[J].上海中医药杂志,2022,56(12):73-78. DOI: 10.16305/j.1007-1334.2022.2202083.
HAN Xinyan,CHEN Ziyu,YUAN Jinfeng,et al.Artemisia annua tea extract attenuating DNCB⁃induced atopic dermatitis by restraining Th2 cell mediated inflammatory responses in BALB/c mice[J].Shanghai Journal of Traditional Chinese Medicine,2022,56(12):73-78. DOI: 10.16305/j.1007-1334.2022.2202083.
目的,2,评价黄花蒿茶(苦草茶)提取物(AT)对2,4-二硝基氯苯(DNCB)诱导的BALB/c小鼠特应性皮炎(AD)是否具有改善皮损作用,并探讨其可能的机制。,方法,2,将32只雄性BALB/c小鼠,分为正常对照组、模型组、苦草茶提取物组及地塞米松组,每组8只。除正常对照组外,其余各组应用DNCB刺激诱导小鼠AD,于第7天灌胃给予相应组模型动物AT(3 g·kg,-1,)和地塞米松(1.5 mg·kg,-1,),连续给药13 d。通过皮肤损伤评分,以及耳厚、耳质量指标考察药物对AD小鼠皮损的作用;利用苏木精-伊红染色法(HE)、甲苯胺蓝(TB)染色法检测小鼠耳组织肥大细胞及嗜酸性细胞浸润;应用酶联免疫吸附法(ELISA)检测血清白介素-13(IL-13)水平;运用定量聚合酶链反应(qPCR)法检测耳组织,IL-13,、胸腺基质淋巴细胞生成素(,TSLP,)、白介素-17(,IL-17,)及肿瘤坏死因子-α(,TNF-α,)等炎症因子的mRNA表达。,结果,2,①与正常对照组比较,反复应用DNCB于小鼠背部及耳部皮肤后,模型组小鼠皮肤出现红肿溃烂、干燥剥落等表现,且小鼠表皮增厚、嗜酸性细胞及肥大细胞浸润增多(,P<,0.05),血清IL-13水平显著升高(,P<,0.05),耳组织,IL-13,、,TSLP,、,IL-17,及,TNF-α, mRNA表达显著上调(,P<,0.05)。②与模型组比较,苦草茶提取物组DNCB诱导的BALB/c小鼠的AD样症状显著减轻,皮肤干燥剥落、红斑、肿胀溃烂现象显著改善(,P<,0.05),耳增厚明显抑制(,P<,0.05),肥大细胞和嗜酸性细胞浸润显著减少(,P<,0.05);小鼠血清IL-13水平显著降低(,P<,0.05),耳组织,IL-13,、,TSLP,、,IL-17,及,TNF-α, mRNA表达显著下调(,P<,0.05)。,结论,2,AT可能通过抑制辅助性T细胞2(Th2)相关炎症细胞因子IL-13、TSLP、IL-17及TNF-α表达,进而对AD产生治疗作用。
Objective,2,To evaluate the effects of Artemisia annua tea extract (AT) on improving skin lesions in dinitrochlorobenzene (DNCB)-induced atopic dermatitis (AD) in BALB/c mice and to explore the possible mechanism.,Methods,2,Thirty-two male BALB/c mice were divided into normal control group, model group, AT group and dexamethasone group, with 8 mice in each group. In addition to the normal control group, AD was induced by DNCB stimulation in each mouse. At the 7th day, the model animal was given intragastric administration of AT (3 g·kg,-1,) and dexamethasone(1.5 mg·kg,-1,)for 13 days continuously. The effects of the drug on AD mice skin lesions were evaluated by clinical skin score, ear thickness and ear weight indexes. The infiltration of eosinophils and mast cells in mouse skin tissue were detected by hematoxylin and eosin (HE) and toluidine blue (TB) staining. ,IL,-,13, level was detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expressions of inflammatory genes such as ,IL,-,13,, thymic stromal lymphopoietin (,TSLP,), ,IL,-,17, and ,TNF,-,α, in ear tissues were detected by qPCR.,Results,2,①Compared with the normal control group, after repeated application of DNCB on the skin of the back and ear of mice, the skin of the model group mice showed erythema, swelling, fester,dryness, flakiness, and other manifestations. The epidermis of the mice was thickened, and the infiltration of eosinophils and mast cells was increased (,P,<,0.05). The serum IL-13 level was significantly increased (,P,<,0.05). The mRNA expressions of ,IL-13,, ,TSLP,, ,IL-17, and ,TNF-α, in ear tissue were significantly up-regulated (,P,<,0.05). ②Compared with the model group, the AD-like symptoms of BALB/c mice induced by DNCB were significantly alleviated by AT treatment. The skin dryness and flakiness, erythema, swelling and fester were significantly improved (,P<,0.05), ear thickening was significantly inhibited (,P<,0.05), and mast cells and eosinophilic infiltration were significantly decreased (,P<,0.05). At the same time, the serum IL-13 level was significantly decreased (,P<,0.05), and the mRNA expressions of ,IL-13,, ,TSLP,, ,IL-17, and ,TNF-α, in ear tissues were also significantly down-regulated (,P<,0.05).,Conclusion,2,AT may have a therapeutic effect on AD by inhibiting the expression of Th2-related inflammatory cytokines including IL-13, TSLP, IL-17 and TNF-α.
特应性皮炎湿疹苦草茶黄花蒿抗炎免疫调节小鼠模型中药研究
atopic dermatitiseczemaArtemisia annua teaArtemisia annuaanti-inflammatoryimmunoregulationmice modeltraditional Chinese herbal medicine research
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