| BackgroundPsoriasis is one of the most chronic inflammatory hyperplastic skin diseases,complex condition,easy to relapse,to patients and families has brought great mental and economic burden disease with a long course and recurrent attacks.The incidence of psoriasis is associated with streptococcal infection,genetic susceptibility,environmental factors,mood,immune disorders The pathogenesis of psoriasis is inadequately understood,a variety of factors such as streptococcal infection,genetic susceptibility,environmental factors,mood and immune disorders are involved in its outbreak,in which the factor of immunity is regarded as the central part.It can lead to a variety of chronic skin inflammatory manifestations.As the immune regulation mainly through the interaction between cytokines to achieve,so the relationship between cytokines and psoriasis has been of concern.A large number of previous studies have shown that IL-23/IL-17 axis plays an important role in psoriasis,but recent studies have found that IL-36 inflammatory pathway may be better than the role of the axis in psoriasis.Elevated levels of IL-36 cytokines and IL-36R were observed in lesions of human psoriatic skin and confirmed in mouse models of psoriasis-like diseases.Mice lacking IL-23,IL-17,or IL-22 were less well protected from disease compared with Il36r-/-mice,indicating an additional distinct activity of IL-36 beyond induction of the pathological IL-23/IL17 axis.Meanwhile,intraperitoneal injection of IL-36 antibody in mice can reduce psoriasis skin lesions.This suggests that IL-36 may play an important role in the pathogenesis of psoriasis.IL-36 cytokines,which are members of the IL-1 superfamily,include three members,namely IL-36a,IL-36?,and IL-36?(originally named IL-1F6,IL-1F8,and IL-1F9,respectively).It has been shown that IL-36 is a strong inducer of the CCR6 ligand CCL20 expression by KC,and IL-36? injection resulted in chemokine expression,leukocyte infiltration,and acanthosis of mouse skin.Meanwhlie,the reports also revealed that IL-36 cytokines is regulated not only by Th17 cytokines,they also potentiate the function of Th17 cytokines.However,the expression of CCL20/CCR6 was positively correlated with IL-17 production.This further suggests that IL-36 may have an interaction with CCL20.In conclusion,IL-36 can promote the activation of Th17 cells and stimulate the secretion of CCL20 from keratinocytes,which may play an important role in psoriasis,so it is very important to further explore the mechanism of IL-36 in psoriasis.In view of foreign research reports,IL-36? was selected as the study object in the study of IL-36? IL-36? in IL-36 family.The expression of psoriasis in mice was induced by imiquimod,and the expression of psoriasis lesions in IL-36? mice was detected by real-time fluorescence quantitative PCR and Western Blot.In addition,intraperitoneal injection of IL-36 antibody,real-time quantitative PCR,immunohistochemical method to study the IL-36 antibody on psoriasis-like lesions and psoriasis-related factors CCL20 and further study the mechanism of IL-36 in psoriasis,psoriasis for the treatment and prognosis to provide a theoretical basis.Objective1.Construction of imiquimod cream-induced psoriasis mouse model.2.The expression of IL-36a in psoriasis-like lesions induced by imiquimod was first explored.3.To investigate whether IL-36 antibody has a mitigating effect on imipramide-induced mouse psoriasis-like inflammation.4.To investigate the effect of IL-36 antibody on CCL20 in psoriasis-induced lesions in mice induced by imiquimod,and to explore the possible mechanism of IL-36 involvement in psoriasis.Methods1.Animal experiment:the establishment of imiquimod cream in mice induced psoriasis lesion model,the control group smear Vaseline cream;2.The expression of IL-36a in the skin of imiquimod mice was detected by real-time fluorescence quantitative PCR and Western blot.3.BALB/c female mice were randomly divided into three groups:vaseline blank control group(short for blank control group),imiquimod model group(short for model group)and IL-36 antibody treatment experimental group(short for experimental group).4.To observe the severity of inflammatory changes of psoriasis in mice,the daily PASI score,HE staining observation of pathological changes in skin lesions,and to measure the thickness of skin lesions.5.The expression of CCL20 in skin lesions was detected by immunohistochemistry and real-time quantitative PCR.Results1.The expression of IL-36a in imiquimod-induced psoriasis lesions in mice and in the control skin in miceReal-time quantitative PCR results showed that the expression of IL-36a mRNA in the model group was significantly higher than that in the control group.(t = 7.523,P<0.05).The Western Blot showed that the expression of IL-36a protein in the lesion of the model group was higher than that of the control group,the difference was statistically significant.(t= 31.695,P<0.05.The results suggest that IL-36a may be involved in the establishment of the psoriasis imiquimod model.2.The effect of IL-36 antibody on imiquimod-induced psoriasis lesions in miceThe results showed that there was no obvious abnormality in the dorsal skin of the blank control group.Erythema appeared on the 3rd day in the model group.After the skin was thickened,the erythema was thickened on the 4th to the 6th day,8 days showed typical psoriasis-like changes,some mice began to appear desquamation,visible point bleeding,film phenomenon;experimental group 3 days part of the mouse back skin is light red,less scaly,less than the model group,the first 5-8 days erythema,scales,thickening increased,but the indicators were less than the model group,the first 8 days did not show typical psoriasis lesions,the indicators were still higher than the blank control group.From the clinical manifestations,this suggests that IL-36 antibodies have a deterrent effect on psoriasis-like lesions in imiquimod mice.HE staining revealed showed that the skin of the control group had no abnormalities on the 8th day.The skin lesions of the model group were changed by psoriasis,the epidermis was thickened and the epidermis was prolonged.The epidermis was more smooth and the number of parakeratosis Reduced thickness is lower than model group.This suggests that IL-36 antibody significantly improves the pathologic severity of psoriasis lesions in imiquimod mice.3.The effects of IL-36 antibody on the expression of CCL20 in psoriasis-like lesions induced by imiquimod in miceThe expression of CCL20 in the model group was mainly in the epidermis,the color was brown,the number of colored cells and the number of the cells were significantly increased compared with the control group(P<0.05).The expression of CCL20 in the control group was significantly lower than that in the control group The CCL20 in the IL-36 antibody group was also mainly located in the epidermis,and the number and number of stained cells were significantly lower than those in the model group.CCL20 staining was light brown.The results of immunohistochemistry showed that the difference between the experimental group and the model group was statistically significant(Z = 3.25,P<0.05).The difference between the experimental group and the control group was not statistically significant(Z = 1.45,P>0.5).In the non-parametric test,we found that CCL20 was mainly expressed in the epidermis of the model group and the experimental group,and there was significant difference between the experimental group,the control group and the model group(P<0.05).Compared with the blank control group,the content of CCL20 in the model group was significantly higher than that in the control group(P<0.05).Real-time quantitative PCR also showed that the difference between the experimental group(1.63 ± 0.19)and the model group(2.65 ±0.47)was significantly different(P<0.05).This suggests that IL-36 antibody significantly reduces the expression of CCL20 in imidocormol-induced mouse psoriatic lesions..Conclusion1.IL-36a may play an important role in the pathogenesis of psoriasis in mice induced by imiquimod,and the expression of IL-36a may be involved in the pathogenesis of psoriasis.2.IL-36 antibody can alleviate imipramine-induced mouse psoriasis-like inflammation,its role may be related to inhibition of CCL20 expression. |
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