Clinical Observation Of The Treatment Of Seborrheic Dermatitis With Cypress Lotion And Its Experimental Mechanism

1 Purpose(1)Observe the clinical efficacy of treating seborrheic dermatitis(damp-heat syndrome)by Jinbai lotion.(2)Optimizing the infection model of Guinea Pig Malassezia to evaluate the fungal effect through grading modelled skin lesion,microscopic examination and fungi culture,etc.(3)Testing the level of model INF-γ、IL-4、IL-2、IL-10,understanding the impact to immune system by Malassezia,discussing the possible functional mechanism of treating Malassezia infected disease by JinBai lotion.(4)The research utilised the cell experiments to observe the impact made by Jinbai lotion and Malassezia on cell proliferation formed by exfoliation.Exploring whether Jinbai lotion exerts its effect of anti-inflammatory and inhibiting the proliferation of epidermis by adjusting the exfoliation to build cellular immune functio.2 MethodThe observation of the effect to treat seborrheic dermatitis(damp-heat syndrome)by Jinbai lotion:The research strictly follows the relevant article and the diagnose standard of seborrheic dermatitis(damp-heat syndrome)provided by the industry standard.The research conducted various experimental plans including random,opened and parallelled comparison between Jinbai lotion and Ketoconazole lotion.The treatment period is four weeks.The research evaluated a general clinical efficacy by observing the therapeutic effect of patient’s erythema,scale,the degree of pruritus and target lesion area.Meanwhile,we recorded any adverse event and recurrence of the disease.Mechanism of JinBai lotionAnimal test(1)Build an Malassezia infected modelAdopting an approach to apply fungus and oil for 7 days to build a Malassezia infected model,observing the model’s erythema and scale and grading them,recording the model’s effect.(2)Observing the change of skin lesion under Guinea Pig Malassezia infected model.Adopting an approach to apply fungus and oil for seven days to build a Malassezia infected model,using Jinbai lotion on the body,while setting up a group contained positive medicine(using Ketoconazole lotion)for comparison,a group included model and a group with nothing.Evaluate the change of skin lesion grades,the percentage of curing fungus,number of fungal culture colonies and pathological changes.(3)ELISA approach to test the content of INF-γ、IL-4、IL-2、IL-10 in group with Jinbai lotion,the group with Ketoconazole lotion,the group with the model and the group with nothing.Cell experiment(1)Build a model to co-cultivate Malassezia and Hacat cell,using MTT approach to the impact to Hacat cell proliferation by Malassezia with different level of concentration.(2)Using MTT approach to test the impact to Hacat cell proliferation by Jinbailotion with different level of concentration.(3)Build a model to co-cultivate Malassezia and cell formed by exfoliation,using Jinbailotion with high,medium and low concentration level to affect the model;set up a group with the model,and a group with nothing.Abstract the supernate from each cultivated group and test the content of IL-8、sICAM-1、TLR-2 by ELIS A.3 ResultClinical efficacyThe clinical efficacy rate of treating seborrheic dermatitis(damp-heat syndrome)by Jinbai lotion was superior to the comparing group,which treated the disease by Ketoconazole lotion.The total efficiency of treatment by Jinbai lotion is 80.56%.However,the comparing group of Ketoconazole lotion is 75.00%.After analyzing by statistics,the data of these two groups has no significant difference(p>0.05).After treatment,the improvement of the grades of skin lesion,pruritus and scale treated by Jinbai lotion was superior than Ketoconazole lotion(p<0.05).With respect to the skin erythema,steatorrhea symptoms and target lesion area,the treatment effect of these two medicines are quite similar(p>0.05).The experimental research(1)Adopting a 7-days fungus-oil applying approach to build Guinea Pig Malassezia infected model.There were two kinds of skin lesion:point shape skin erythema,skin erythema and a significant amount of oil scale.The skin applied fungus and oil generated obvious skin erythema and scale after three days.It became most apparent in the 5th day.Erythema and some chipped scale appeared on the skin applying the 7-day fugus and oil approach.It became the most evident on three days after stopping using the fungus and oil.Concerning the grades of infected skin lesion,the average skin lesion value applying fungus and oil was higher than the one that only applied fungus(p<0.05),and it lasted for a longer time.Under the fungal microscopic examination,the animal in two groups got 100%positive scale cultivation text 4 days after stopping applying fungus.21 days after applying fungus,the positive rate of microscopic examination under fungus-oil application approach was apparently higher than the group merely applied fungus(p<0.05).(2)Bacteriostasis experiment of JinBai lotionSkin integral aspect:as the continuation of treatment time,JinBai lotion,Ketoconazole lotion group,model group,all skin lesion score showing a downward trend,JinBai lotion group with ketone health zun lotion skin lesion area decreased statistically significant(p<0.05).The score of guinea pig skin loss was the fastest and the overall score was lower.The cure rate of the fungus of JinBai and Ketoconazole lotion group were 70%,which was significantly different from that of the model group(p<0.01).Fungi cultivation colony count:JinBai lotion group,Ketoconazole lotion number fungal colony culture with the model group with significant difference(p<0.05),ketoconazole colony count<JinBai lotion group<model group.(3)Levels of INF-γ、IL-4、IL-2、IL-10In the model group,IL-2 and INF-ylevels were compared with other groups decreased significantly.There was no statistical significance(p>0.05)between JinBai lotion group and k Ketoconazole lotion group.The content of IL-4 and IL-10 in guinea pig serum of the model group was higher than that in the blank group and the ketoconazole group increased significantly(p<0.05).There was no statistical significance(p>0.05)between JinBai lotion group and k Ketoconazole lotion group.(4)The effect of malassezia on the proliferation of keratinocytes.At 24h,(fungi/cells =30/1)co-cultured keratinocytes proliferation was significantly inhibited,and keratinocytes in the remaining experimental groups were well developed.At 48h and 72h,(fungi/cells =20/1),(fungi/cells =30/1),the co-culture pores were compared with the other groups,and their proliferation was significantly inhibited(P<0.05).(5)The effect of JinBai lotion on the proliferation of keratinocytes.In addition to the low concentration group 24h,all concentrations of JinBai lotion showed inhibition of keratinocytes proliferation at the observation point(p<0.05),and the inhibition degree was positively correlated with the concentration size.The cytotoxicity of the high concentration group was 2,and the cytotoxicity was 1.(6)The content of IL-8,sICAM-1 and TLR-2 that are secreted and expressed by keratinocytes.There was a significant increase in the secretion of IL-8,sICAM-1 and TLR-2(P<0.01)with keratinocytes in the co-culture of the cultured cells.Compared with the model group,the secretion level(P<0.01)was significantly reduced in different concentrations of JinBai lotion(P<0.01),and the reduction level was positively correlated with the drug concentration.4 ConclusionThe clinical research showed that:It is safe to treat seborrheic dermatitis(damp-heat syndrome)by Jinbai lotion.The lotion can effectively relieve skin erythema,scale,pruritus,steatorrhea symptoms,and decrease the area of the skin lesion.The experiments suggests that:(1)Jinbai lotion has a good effect on fungi stasis.(2)The Guinea Pig Malassezia infected model contains Th1/Th2 drifting.Jinbai lotion might balance these two to reach the anti-inflammatory and anti-bacterial effect.(3)The Malassezia with different legal of concentration can inhibit exfoliation,and even make its apoptosis.Meanwhile,the Malassezia can stimulate the cell formed by exfoliation to secrete cell factor.Jinbai lotion might cut off the exfoliation and form cell TLR-2 receptor expression,further,inhibit the T-cell activation and reduce the secretion mechanism of IL-8 and sICAM-1 to relieve the react of local inflammation,therefore to inhibit the epidermal growth.