| Objective1.To acquire several effective kings of Chinese medicine for Aropic dermatitis from previous literatures by systematic Meta-analysis.According to Chinese medicial erperts’ recommendation,Portulaca oleracea L.and ADHO1 are confirmed to be studied through next prelimitary animal experiment.2.To establish the atopic dermatitis murine model by DNFB sensitization and excitation on C57BL/6 mice3.To observe the efficacy of the external application of Portulaca oleracea L.and ADH01 for atopic dermatitis and their impact on the immunological function and filaggrin expression.To disscuss their possible mechanism on the TH1/TH2 regulation and repair of skin barrier.To provide a basis for developing new effective external preparation for atopic dermatitis.Methods1.According to the retrieving strategy,randomized controlled trials(RCTs)external therapies of traditional Chinese medicine for atopic dermatitis from January 1989 to 2015 were retrieved from the database of CNKI,WANFANG,VIP,sinomed,PubMed,Web of Science,and the outcomes of which were assessed systematically by meta-analysis with the RevMan5.2,the quality of studies were assessed by Jadad scale and the criteria recommended Cochrane Collaboration Handbook 5.1.Analze the formula in the studies which were filtrated and consult my tutor and other clinical Chinese medicial spelicialists.2.Ten female 7-week-old C57BL/6 mice were randomly divided into 2 groups:model group and control group.The mice’ s back hair were shaved 1 day prior to sensitization.In the first day,50 μ 1 and 20 μ 1 o.5%DNFB in acetone/olive oil(4:1)were evenly respectively applied to the dorsal skin and right ear to make the first sensitization.Since the fifth day,0,2%DNFB have been applied to the same area every two days with the same volume to make the excitation.Then the excitation was conducted once a week to keep the skin lesion The same volume of acetone/olive oil vehicle was applied instead of DNFB solution to control group.On the 29th day,record the skin lesion signs,scratching frequency in ten minutes and ear thickness.Then save the serum and the skin lesion tissue.To detect serum IgE、IL-4and IFN-γlevels with ELISA assay.The skin lesion tissue was fixed with 10%methanol and made into pathological section.To detect the IL-4 and IFN-γmRNA expression from skin lesion Cells with Real-time quantitative RT-PCR.To detect the Mean Optical Density of the filaggrin from lesion with immunohitochemical assay.3.Thirty female 7-week-old C57BL/6 mice were randomly divided into 6 groups:the model group,the cortisone group(0.1%hydrocortisone butyrate),low and high dosage of Portulaca oleracea L.water extract group(n=5),and low and high dosage of ADHO1 water extract group(n=5).According to previous methods,the mice were given external DNFB to become the AD model.Since the excitation stage,The recommended external dosage of Portulaca oleracea L.water extract.ADH01 water extract and 0.1%hydrocortisone butyrate were given to the corresponding groups.The model group was given nothing.4 weeks later,record the skin lesion signs and evaluate the severity of skin inflammation,scratching frequency in ten minutes and ear thickness.Then save the serum and the skin lesion tissue.To detect serum IgE、IL-4and IFN-γlevels with ELISA assay.The skin lesion tissue was fixed with 10%methanol and made into pathological section.To detect the IL-4 and IFN-γmRNA expression from skin lesion Cells with Real-time quantitative RT-PCR.To detect the Mean Optical Density of the filaggrin from lesion with immunohitochemical assay.Results1.17 studies which were included showed that the effective rate of experimental group was better than that of control group[OR=1.79,95%CI(1.27,2.80),P<0.05];12 studies showed that the cure-rate of experimental group was better than that of control group[OR=1.65,95%CI(1.27,2.15),P<0.01];9 studies and another 5 studies showed that the Incidences of adverse effect and the recurrence rate of experimental group were both lower than that of their respectively control group[OR=0.23,95%CI(0.11,0.47),P<0.01;0R=0.14,950%CI(0.05,0.37),P<0.01].According to the anysis of the formula from the studies which were filtrated and the tutor and other clinical Chinese medicial spelicialists’ advise,the external application of Portulaca oleracea L.and ADHO1 water extract were confirmed to be given to the AD murine model with clinical common dosage and the way of bathing.2.mice of the model group appeared erythema,exudation,skin exfoliation and lichenification on the dorsal skin and obvious right ear swelling.Histopathology showed hyperkeratosis,spinous layer thickening,significant infiltration of plenty of lymphocytes in the model group.Compared with the control group,the scratching behavior frequency and ear thickness in the model group obviously increased(P<0.05);the Serum IgE and IL-4 level in the model group obviously increased(P<0.05);the IL-4 mRNA expression of skin lesion in the model group significantly increased(P<0.05);the filaggrin expression of skin lesion in the model group obviously decreased(P<0.05).3.Compared with the model group,the skin inflammmation in the Portulaca oleracea L.group.the ADHO1 group and the cortisone group appeared slighter erythema,skin exfoliation,lichenification and no exudation.Histopathology showed slighter spinous layer thickening and infiltration of lymphocytes in the Portulaca oleracea L.group、the ADH01 group and the cortisone group,which also showed hyperkeratosis.Compared with the model group,the skin leision severity score decreased significantly in the other five groups(P<0.0125);the scratching behavior frequency obviously decreased in the Portulaca oleracea L.group、the high dosage of ADHO1 group and the cortisone group(P<0.05);the ear thickness obviously decreased in the other five groups(P<0.0125);the serum IgE level obviously decreased in the Portulaca oleracea L.group、the high dosage of ADHO1 group and the cortisone group(P<0.05);the serum IL-4 level obviously decreased the Portulaca oleracea L.group and the cortisone group(P<0.05);the serum IFN-γlevel obviously decreased in the cortisone group(P<0.05),the serum IFN-γlevel show no difference in the Portulaca oleracea L.groups(P>0.05);the skin lesion IL-4mRNA expression obviously decreased in the cortisone group and the high dosage of Portulaca oleracea L.group(P<0.0125);the skin lesion IFN-γ mRNA expression obviously increased in the high dosage of Portulaca oleracea L.group(P<0.0125),obviously decreased in the cortisone group(P<0.125);the skin lesion FLG optical density value obviously increased in the high dosage of Portulaca oleracea L.group and the high dosage of ADHO1 group(P<0.05).Compared with the cortisone group,the scratching behavior frequency increased with obvious difference in the Portulaca oleracea L.group and the high dosage of ADHO1 group(P<0.05);the ear thickness increased with obvious difference in the Portulaca oleracea L.group and the ADHO1 group(p<0.0125);the serum IgE level increased with obvious difference in the low dosage of Portulaca oleracea L.group(P<0.05).Compared with the low dosage of Portulaca oleracea L.group,the skin leision severity score showed no obvious difference in the high dosage of Portulaca oleracea L.group and the ADHO1 group(P>0.0125),the scratching behavior frequency obviously decreased in the high dosage of Portulaca oleracea L.group(P<0.05);the ear thickness obviously decreased in the high dosage of Portulaca oleracea L.group and the high dosage of the ADHO1 group(p<0.05);the serum IgE level obviously decreased in the high dosage of Portulaca oleracea L.group(p<0.05);Compared with the high dosage of Portulaca oleracea L.group,the scratching behavior frequency obviously decreased in the high dosage of ADHO1 group(P<0.05),the ear thickness obviously increased in the ADHO1 group(p<0.05),the serum IgE level obviously decreased in the high dosage of ADHO1 group(p<0.05);Compared with the low dosage of ADHO1 group,the ear thickness obviously increased in the high dosage of ADHO1 group(p<0.05).Conclusion1.The Meta analysis result proved that compared with the western medicial treatment,external therapies of TCM for AD has more effective efficacy、lower Incidences of adverse effect and the recurrence rate.Analze the formula in the studies which were filtrated and consult my tutor and other clinical Chinese medicial spelicialists.This study objectively assess the efficacy and safety of external therapies of TCM for atopic dermatitis(AD)and offer evidence for EBM.According to the anysis of the formula from the studies which were filtrated and the tutor and other clinical Chinese medicial spelicialists’ advise,the external Chinese medicine were confirmed to be conducted futher assement and filtrating.2.The Atopic dermatitis murine model was successfully established by DNFB in the C57BL/6 mice with significant changes on the skin inflammation,pruritus,seroimmunological indexes,skin lesion immunological indexes and filaggrin expression,which are all similar with human atopic dermatitis.This model could be used for the study of AD in many aspects such as pathogenesis,treatments and transgenosis3.Though futher assement and filtrating,the high dosage of Portulaca oleracea L.and ADHO1 could both obviously decrease the severity of skin lesion,relieve itching,decrease the serum IgE level and promote the filaggrin expression and repair the skin barrier.The Portulaca oleracea L.also could regulate the TH1/TH2 unblance.Portulaca oleracea L.and ADHO1 all have great effection for AD and suit futher development.Future study could focuse on the immunoregulation relatived study with preparation having more propotion of ADH01 spore.External preparation could take other new dosage form for best response. |
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