摘要
目的探讨二妙丸对特应性皮炎小鼠的缓解作用。方法体内实验:将BALB/c小鼠随机分为正常组、模型组、地塞米松组(2 mg/kg)和二妙丸高、中、低剂量组(4.68、2.34、1.17 g/kg),采用反复涂抹DNCB溶液构建特应性皮炎小鼠模型,造模同时灌胃给药,连续21 d,观察并记录小鼠背部皮损状况、耳肿胀度、耳片重量差,HE染色观察皮损组织病理变化,甲苯胺蓝(TB)染色观察皮损组织肥大细胞浸润,IHC法检测皮损组织巨噬细胞标志物F4/80表达,ELISA法检测血清TSLP、IL-4、IL-5和总IgE水平。体外实验:取对数生长期RAW264.7细胞,给予400、200、100μg/mL二妙丸进行干预,CCK-8法检测细胞增殖情况,Griess法检测细胞上清液NO水平,ELISA法检测细胞上清液TNF-α、IL-1β、IL-6水平,Western blot法检测细胞MAPK/NF-κB信号通路相关蛋白表达。结果体内实验:与模型组比较,二妙丸高剂量组小鼠背部皮损评分、右耳肿胀度、左右耳片重量差均降低(P<0.05,P<0.01),皮损组织表皮厚度减小,肥大细胞、巨噬细胞浸润减轻(P<0.05,P<0.01),血清炎症因子TSLP、IL-4、IL-5、总IgE水平降低(P<0.05,P<0.01),皮损组织F4/80表达降低(P<0.01)。体外实验:与模型组比较,二妙丸400、200μg/mL组细胞NO、TNF-α、IL-1β、IL-6水平降低(P<0.05,P<0.01),P38、JNK和P65蛋白磷酸化水平降低(P<0.05,P<0.01)。结论二妙丸能够缓解DNCB诱导的特应性皮炎小鼠皮损炎症,其机制可能与抑制巨噬细胞MAPK/NF-κB信号通路激活,减轻巨噬细胞浸润和降低Th2细胞因子有关。
AIM To explore the relieving effect of Er Miao Wan on atopic dermatitis in mice.METHODS In vivo experiment:BALB/c mice were randomly divided into normal group,model group,dexamethasone group(2 mg/kg)and high,medium and low dose groups of Er Miao Wan(4.68,2.34 and 1.17 g/kg).The mouse model of atopic dermatitis was established by repeatedly smearing DNCB solution,and the model was given orally for 21 days.The skin lesion condition on the back of mice,ear swelling degree,and the weight difference between ear lobes were observed and recorded.HE staining was used to observe the histopathological changes in the skin lesion tissues of mice.Toluidine blue(TB)staining was used to observe the infiltration of mast cells in skin lesions.The expression of macrophage marker F4/80 in skin lesions was detected by IHC.The serum levels of TSLP,IL-4,IL-5 and total IgE were detected by ELISA.In vitro experiment:RAW264.7 cells in logarithmic growth period were given 400,200 and 100μg/mL Er Miao Wan for intervention.Cell proliferation was detected by CCK-8 method.NO level in cell supernatant was detected by Griess method.TNF-α,IL-1βand IL-6 levels in cell supernatant were detected by ELISA method.The expressions of proteins related to the MAPK/NF-κB signaling pathway in cells was detected by Western blot.RESULTS In vivo experiment:Compared with the model group,the scores of back skin lesions,the swelling degree of right ear and the weight difference between left and right ear pieces in the high-dose group of Er Miao Wan decreased(P<0.05,P<0.01),the thickness of skin lesions decreased,the infiltration of mast cells and macrophages decreased(P<0.05,P<0.01),and the inflammatory factors TSLP,IL-4,IL-5 and total IgE levels in serum decreased(P<0.05,P<0.01),and the expression of F4/80 in the skin lesions decreased(P<0.01).In vitro experiment:Compared with the model group,the levels of NO,TNF-α,IL-1βand IL-6 in Er Miao Wan 400 and 200μg/mL groups decreased(P<0.05,P<0.01),and the phosphorylation levels of P38,JNK and P65 proteins decreased(P<0.05,P<0.01).CONCLUSION Er Miao Wan can alleviate skin lesion inflammation in DNCB-induced atopic dermatitis mice,and its mechanism may be related to inhibiting the activation of MAPK/NF-κB signaling pathway of macrophage,reducing macrophage infiltration and reducing Th2 cytokines.
作者
杨胜晋
刘燕娇
鲁诚
邓世俊
李婧
张欣佳
张祎
王睿睿
张丽娟
YANG Sheng-jin;LIU Yan-jiao;LU Cheng;DENG Shi-jun;LI Jing;ZHANG Xin-jia;ZHANG Yi;WANG Rui-rui;ZHANG Li-juan(Yunnan University of Chinese Medicine,Kunming 650500,China)
出处
《中成药》
北大核心
2025年第11期3591-3600,共10页
Chinese Traditional Patent Medicine
基金
国家自然基金地区基金项目(81760741,82160782)
云南省科技厅中医联合专项重点项目(202301AZ070001-006)
云南省科技厅中医联合专项面上项目(202101AZ070001-187)
云南省匡海学专家工作站(202305AF150029)
云南中医药大学中药学院研究生自主创新研究项目(30271108100)。
