摘要
目的:探讨4种中药润肤霜对2,4-二硝基氟苯(DNFB)致特应性皮炎小鼠的治疗作用及机制。方法:BALA/c小鼠按体质量随机分成正常组、模型组、空白膏霜组(润肤霜A组),芍药甘草汤组(润肤霜B组),芍药甘草汤加马齿苋,人参和忍冬藤组(润肤霜C组),芍药甘草汤加人参和忍冬藤组(润肤霜D组),给药剂量均为25 g·kg-1,以及阳性药他克莫司软膏组(3 g·kg-1),每组10~12只。除正常组外,其余各组用0.5%DNFB涂抹小鼠背部脱毛皮肤,每只100μL,连续7 d。第7天起,各组给予相应润肤霜外用干预,1次/d,连续15 d,正常组和模型组不给药。每周称量动物体质量1次,每周对动物背部皮肤进行评分3次,记录皮损状况分值;处死小鼠后,取左右耳,打孔称量双耳质量并计算肿胀度;取背部皮肤固定,苏木素-伊红(HE)染色进行病理组织学检查,观察小鼠背部皮肤组织病理改变并评分;末次给药后取血,酶联免疫吸附测定(ELISA)检测血清中免疫球蛋白E(IgE)含量;蛋白免疫印迹法(Western blot)检测皮肤组织中信号传导与转录激活因子3(STAT3),磷酸化(p)-STAT3蛋白的表达。结果:与正常组比较,模型组小鼠体质量持续性降低,造模区出现红斑、水肿、干燥、脱屑、结痂剥脱等炎性改变,皮损状况分值显著增加;耳部皮肤角质层增厚,耳肿胀度明显增加;镜下见表皮局部坏死脱落、表皮增厚、表皮增生,角质层的角化过度和角化不全,以及皮下组织炎症细胞浸润等病变;血清IgE含量显著性增高;皮肤组织中p-STAT3的表达增加。与模型组比较,润肤霜C和D组小鼠体质量明显增加(P<0.01),皮损状况评分明显降低(P<0.05,P<0.01);润肤霜B,C,D组较模型组小鼠耳廓肿胀度显著减少(P<0.01);润肤霜C组小鼠背部皮肤坏死缺损和表皮增生的病变程度较模型组明显减轻(P<0.05,P<0.01);润肤霜C和D组小鼠血清中IgE含量较模型组明显降低(P<0.05,P<0.01);润肤霜C组小鼠皮肤组织中p-STAT3蛋白表达明显低于模型组(P<0.05)。结论:润肤霜B,C,D对特应性皮炎均有一定的治疗效果,其中润肤霜C的治疗作用最为明显,作用机制可能与其抑制血清IgE含量的升高,及抑制STAT3的磷酸化,从而降低炎性细胞因子的水平有关。
Objective: To investigate the therapeutic effect and possible mechanism of four types of Chinese herbal moisturizers made in laboratory for atopic dermatitis induced by 2,4-dinitrofluorobenzene(DNFB)in mice. Method:According to the body weight,BALA/c mice were randomly divided into normal group,model group,blank cream group(moisturizer A),Shaoyao Gancaotang group(moisturizer B),Shaoyao Gancaotang with Portulacae Herba,Ginseng Radix et Rhizoma and Honeysuckle Stem group(moisturizer C),and Shaoyao Gancaotang with Ginseng Radix et Rhizoma and Honeysuckle Stem group(moisturizer D),with the dose of 25 g·kg-1 per day,as well as tacrolimus ointment group of 3 g·kg-1 per day,with 10 to 12 mice in each group. Except the normal group,the mice in the other groups were treated with 0.5%DNFB in the hair removal skin of back,100 μL each for 7 days. Starting from the 7 thday,each group was given the appropriate skin cream for external use intervention,once per day,for 15 consecutive days,except for the normal and the model groups. The animal body mass was measured once a week,and the animal back skin was graded three times a week,and the skin lesion score was recorded. After the mice were killed,the left and right ears were taken,the weight of both ears was punched and the degree of swelling was calculated. The back skin was fixed and stained with hematoxylin-eosin(HE)method,and then pathologic examination was conducted to observe and score the pathological changes of mouse back skin. Blood was obtained after the last dose and enzyme-linked immunosorbent assay(ELISA)was used to determine the immunoglobulin(Ig)E content in serum. Western blot was used to measure the expression of signal transduction and activator of transcription 3(STAT3),phosphorylation(p)-STAT3 in the skin tissue. Result: Compared with the normal group,the body mass decreased continuously, a series of inflammatory changes such as erythema, edema, dryness,desquamation and callus exfoliation and so on occurred in the modeling area,and the skin lesion score increased significantly in the model group. Additionally,the cuticle of ear skin was thickened and the degree of ear swelling was obviously increased in the model group. Microscopically,the occurred changes in the model mice included the local necrosis of the epidermis, epidermal thickening, epidermal hyperplasia, and the hyperkeratosis and hypokeratosis in the cuticle,as well as the subcutaneous inflammatory cell infiltration and so on. Furthermore,the content of serum IgE andthe expression of p-STAT3 in skin tissues increased significantly in the model group. Compared with the model group,the body mass of mice in group C and D was significantly increased(P<0.01),and the skin lesion status score was decreased(P<0.05,P<0.01). The degree of auricle swelling was significantly reduced in group B,C and D compared with that in the model group(P<0.01). The degree of skin necrosis and defect and epidermal hyperplasia of mice in moisturizer C group was significantly reduced compared with that in model group(P<0.05,P<0.01). Serum IgE levels of mice in group C and D were significantly lower than those in the model group(P<0.05,P<0.01). The expression of p-STAT3 protein in skin tissues of mice in moisturizer C group was significantly lower than that in model group(P<0.05). Conclusion:The moisturizers B,C and D all have certain therapeutic effect on atopic dermatitis,among which moisturizers C has the most obvious therapeutic effect. The possible mechanism may be that it reduces the level of inflammatory cytokines by inhibiting the increase of serum IgE content and the phosphorylation of STAT3.
作者
王海林
刘陆
回连强
刘婷
李春
李成亮
曹春雨
WANG Hai-lin;LIU Lu;HUI Lian-qiang;LIU Ting;LI Chun;LI Cheng-liang;CAO Chun-yu(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 450000,China;Henan University of Chinese Medicine,Zhengzhou 450000,China;LB Cosmeceuticals Technology(Shanghai)Co.Ltd.,Shanghai 200233,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第22期7-15,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
北京市自然科学基金项目(7132152)。
关键词
特应性皮炎
芍药甘草汤
中药润肤霜
白芍总苷
甘草总黄酮
atopic dermatitis
Shaoyao Gancaotang
traditional Chinese medicine moisturizers
total Paeoniae Radix Alba glycosides
total Glycyrrhizae Radix et Rhizoma flavonoids
