| Objective: Atopic dermatitis is a chronic inflammatory itchy skin disease.It begins from the infantile period with high incidence,and seriously affects the patients’ quality of life.Its pathogenesis is related to scratching stimulation,microbial colonization and skin barrier damage.For the treatment of atopic dermatitis,it is difficult to maintain long-term use of oral medicine due to sleepiness,drowsiness and other adverse effects.Western topical agents are often limited by adverse effects or high price.Therefore,topical treatment with traditional Chinese medicine has great significance in treatment of atopic dermatitis.The transdermal absorption is a key factor for topical use of traditional Chinese medicine.Badger oil,a traditional Chinese medicine with strong penetration,is a good transdermal absorption carrier.It can promote wound healing,is probable able to repair the skin barrier.Therefore,in this study,we made-up the “self-made badger oil liniment” using badger oil and phellodendron,honeysuckle,Fangfeng from the representative traditional formula for atopic dermatitis.We try to elucidate the therapeutic effects,side effects and the mechanism of the “self-made badger oil liniment”,in order to provide evidence for its use as a new traditional Chinese medicine preparation with good therapeutic effect,lesat side effect and reasonable price.Methods: First,we establish the mice model of atopic dermatitis.Twenty SPF grade female BALB / c mice of 6-8 weeks were purchased and randomly divided into four groups: blank control group(without modeling),model group(killed after successful modeling),negative control group(without topical treatment after modeling),positive control group(topical hydrocortisone butyrate)and treatment group(topical Badger oil liniment).The back of the mice were depilated and repeatedly sensitiazed by ovalbumin.The model was verified by pathological changes of the skin and detection of total serum Ig E.Then the therapeutic effects and adverse effects of self-made badger oil liniment were observed.After extablishment of AD model,mice of each group were given respective topical agents twice a day.One week later,the improvement of the lesions on the back of the mice and the adverse effects were abserved by naked eye,the pathological changes were observed after biopsy,and the total serum Ig E of each mouse was detected by ELISA.Then we explore the effects and mechanism of self-made badger oil liniment on the skin barrier.The skin barrier of mice in each group was observed through HE stained histological slides.The expression of FLG was detected by immunohistochemistry and real-time PCR,respectively.Finally,Staphylococcus aureus was cultured in vitro,and the inhibitory effect of self-made badger oil liniment and its components on Staphylococcus aureus was observed by paper method of bacteriostatic test.Results: Compared with the blank group,mice in modeling group developed erythema,edema,dryness,erosion,bleeding and crusts on the back.Pathological findings of the lesions of mice in modeling group showed hyperkeratosis with parakeratosis,crusting,intercellular edema,intercellular bridge,and lymphocyte infiltration around the blood vessels.The total serum Ig E of mice in modeling group was higher than that in the blank group.The difference was statistically significant(P < 0.05).After treatment,the negative control group revealed a little improvement of the lesions,but still edema and obvious desquamation,intercellular edema on histological findings.The positive group revealed improvement of the lesions,although still some desquamation.Histologial findings showed less edema but obviously thinner epidermis.The treatment group revealed improvement of the lesions,although still some desquamation.No side effects including erythema,edema were observed.Histological findings showed less intercellular edema,without obvious change in thickness of the epidermis.The total serum Ig E of treatment group and positive control group was decresed compared with the negative control group,with statistical significance(P < 0.05).Histological observation of the skin barrier indicated hyperkeratosis and epidermal thickening of the modeling group,thinner epidermis of the negative control group than the modeling group,obviously thinner epidermis of the positive control group than the blank group.While the epidermis in the treatment group is thinner than the modeling group,but thicker than the blank group,suggesting barrier impairment of the modeling group and positive control group,and improvement of the barrier impairment of the treatment group.Immunohistohemistry and staining area analysis revealed that the FLG of the blank group was mainly expressed in the orneum,expression of FLG in the modeling group and negative control group was decreased,expression of FLG in the treatment group was increased compared with the negative control group.The differences above were statistically significant(P<0.05).The result of real-time PCR was in accordance with that of the immunohistochemistry,suggesting that the effect of self-made badger oil liniment on skin barrier improvement might be associated with increased FLG expression.Finally,the results of the bacteriostatic test revealed that the phellodendron amurense and self-made badger oil liniment had obvious antibacterial effect on Staphylococcus aureus in vitro,with the diameter of antibacterial circle 19.00 ± 0.71 mm and 10.11 ± 0.78 mm respectively,while fangfeng,honeysuckle and badger oil had no inhibitory effect on Staphylococcus aureus.Conclusion: Self-made badger oil liniment has therapeutic effect on mouse atopic dermatitis model,without obvious side effects.The mechanism might be associated with decrease of serum Ig E level,improvement of sin barrier by promoting FLG expression,and inhibition of Staphylococcus aureus. |
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