| The term of dermatitis-eczema refers to a broad range of common allergic skin diseases,including allergic contact dermatitis (ACD), eczema and atopic dermatitis (AD). Because ofits high prevalence, protracted course and often accompanied by serious itching, it affects thepatients quality of life a lot. Nowadays, drug treatment for the disease is mainly concentratedin symptomatic treatment using anti-histamine drugs systemically and corticosteroidstopically. Owing to its complex etiology and pathogenesis, long course of treatment, beingeasily relapsed, besides the potential side effects of long-term use of these drugs, new drugsfor dermatitis-eczema have been in need badly.Traditional Chinese medicine has unique advantages in the treatment of dermatitis-eczema,which can effectively expelling wind, promoting urination, removing heat, moisturizingdryness and anti-allergy, through a variety of ways. Traditional Chinese medicine thinkshighly of the synergy effect of various drugs, as well as the balance overall. Therefore, it mayhave more efficacy and fewer side effects on dermatitis-eczema, as compared withanti-histamine drugs and corticosteroids. Shenshe lotion is composed of Radix SophoraeFlavescentis, Fructus Cnidii, Cortex Phellodendri, Radix Angelicae Sinensis, RhizomaChuanxiong, Radix et Rhizoma Rhei and Broneolum Syntheticum. It may have effects ofantibacterial, anti-inflammatory, expelling wind, moisturizing dryness, cooling blood andpromoting blood flow. Being administered through the skin not only can avoid the irritationof gastrointestinal tract by oral administration, decrease drugs concentration in blood, reduceside effects, but also can improve patients’ compliance. It is expected to be a new clinicalpraeparatum for the treatment of dermatitis-eczema.A lot of animal models in mice, rats and guinea pigs have been made in the research ofdermatitis-eczema. These models did provide good platforms and tools for the explorer ofetiology, pathogenesis, therapeutic effect and drug development for dermatitis-eczema. Thetwo of the most classic models were mice model with ACD for the acute phase of diseases,and mice model with chronic dermatitis-eczema for the chronic phase.In this study, we try to explore the effect of Shenshe lotion on dermatitis-eczema withacute and chronic dermatitis-eczema mice models. We also try to explore its possiblemechanism by study the expression of some inflammation-related cytokines and chemokinein both mRNA level and protein level, so as to provide experimental basis for its clinic use later.Part I The effect of Shenshe lotion on mice model of ACDIn this study,2,4-dinitrofluorobenzene (DNFB) was used for sensitization on abdomenand stimulation on ears in BALB/c mice to establish ACD models.60BALB/c mice wererandomly divided into six groups: blank control group, model control group, high to low doseShenshe lotion group (1:2,1:1and1:0.5) and the positive control group (1%hydrocortisonesolution). Mice were stimulated on ears with1%DNFB following the abdominal sensitization3days ago, and12h after that, they were given different treatment respectively. Ear thickness,allergic effect and scratching index were calculated before and12h,48h after the stimulation,and pathological changes such as inflammatory cell infiltration in ear were observed byhematoxylin eosin (HE) staining.It was showed that Shenshe lotion of different concentrations could reduce the earthickness and score of allergic effect, ease the scratching phenomenon, and reduce the degreeof inflammation of the ear skin. There was no difference between Shenshe lotion and1%hydrocortisone solution in general. While1%hydrocortisone solution did better in reducingthe degree of inflammation, and Shenshe lotion better in easing the scratching. It wasindicated that Shenshe lotion might have a good therapeutic effect on ACD model as1%hydrocortisone solution in general, without a dose dependent phenomenon.1%hydrocortisone solution did better in reducing the degree of inflammation, while Shenshelotion did better in anti-itching.Part Ⅱ The effect of Shenshe lotion on mice model of chronic dermatitis-eczemaIn this study,2,4-dinitrochlorobenzene (DNCB) was used for sensitization on abdomenand repeated stimulation on ears in BALB/c mice to establish chronic dermatitis-eczemamodels.60BALB/c mice were randomly divided into six groups: blank control group, modelcontrol group, high to low dose Shenshe lotion group (1:2,1:1and1:0.5) and the positivecontrol group (1%hydrocortisone solution). Mice were stimulated on ears repeatedly per3-day with20μl0.5%DNCB for four times following the abdominal sensitization with100μl7%DNCB5days ago, and12h after the first stimulation, they were given different treatmentrespectively. Ear thickness, allergic effect and scratching index were calculated before thefirst stimulation,24h after each stimulation and3days after the last stimulation, andpathological changes such as inflammatory cell infiltration in ear were observed by HEstaining.It was showed that Shenshe lotion of different concentrations could reduce the ear thickness and score of allergic effect, ease the scratching phenomenon, and reduce the degreeof inflammation of the ear skin.1%hydrocortisone solution did better in reducing the earthickness, score of allergic effect and degree of inflammation than Shenshe lotion, meanwhile,Shenshe lotion did better in easing the scratching. It was indicated then, Shenshe lotion mighthave a good therapeutic effect on chronic dermatitis-eczema, especially in relieving pruritus.Part Ⅲ Exploring the anti-inflammatory mechanism of Shenshe lotion on mice modelsOn the basis of the pharmacodynamic research, we explored the anti-inflammatorymechanism of Shenshe lotion on dermatitis-eczema, with the serum and ear samples collectedabove. We detected protein levels of TNF-α, IFN-γ, IL-1β, IL-4and CCL27in peripheralblood with ELISA, and their mRNA levels in ear lesions with realtime PCR.It was found that when ACD and chronic dermatitis-eczema models were built in mice, thelevels of TNF-α and IFN-γ were significantly decreased, while IL-1β and IL-4levels wereincreased in peripheral blood, which showed a typical Th2immune response. As to themRNA levels in lesions, TNF-α, IFN-γ, IL-1β, IL-4and CCL27were all significantlyincreased, showing a significant inflammatory response. There was no change in peripheralblood of ACD mice after the treatment of Shenshe lotion, meanwhile, IL-4levels decreased inperipheral blood of dermatitis-eczema models treated with high concentrations of Shenshelotion. And their mRNA levels were all significantly decreased in lesions of both ACD andchronic dermatitis-eczema models, with no significant difference among differentconcentration of Shenshe lotion group, showing an improvement of the inflammation inlesions. It was showed, by immunohistochemistry, that CCL27protein level was significantlyhigher in lesions of mice models with ACD and chronic dermatitis-eczema, and after thetreatment with Shenshe lotion and1%hydrocortisone solution, CCL27protein level wasdecreased significantly, especially in epidermis, showing a positive correlation with theimprovementof the inflammation in lesion. It was indicated that, Shenshe lotion might play itsanti-inflammatory effects mainly by affecting TNF-α, IFN-γ, IL-1β, IL-4, CCL27and otherinflammatory factors in local lesions, other than those in serum. And further study wasrequired to explore its specific pathways and molecular mechanisms. |
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