| ?Background?Vasculitis is a term involves a cluster of diseases which stems from inflammation and necrosis within or around vascular endothelial cells.In pathological aspect,it may show a series change from pure sight necrosis to granuloma,during which process the inflammation cells can be neutrophil leukocyte,eosinophilic leukocytes,lymphocyte and histocytes.The classification of vasculitis is a little complex,there has been no accordant standard so far.Yet doctors generally differentiate this group of diseases according to the diameter of encroached blood vessels or the type of infiltrative cells.Being the most common type of vasculitis in dermatosis clinic,Leukocytoclastic vasculitis(LCV)involves various diseases including Henoch-Schonlein purpura(HSP),allergic vasculitis(ACV),urticarial vasculitis(UV).The inducement of LCV usually relates to drug,inflammation,food,chemical substance,systemic disease and so on while the mechanism usually to antigen,antibody,alexin,endotheliocyte,inflammation cell,various inflammatory factors and so on.Most researchers believe that LCV must be relevant to type III hypersensitivity,it is the chain reaction between circulating immunocomplex,complement,neutrophil leukocyte plays the core role of the mechanism of immune complex type of vasculitis.Other researchers demonstrate that antinenutrophil cytoplasmic antibody(ANCA)may involve the development of LCV.Recent years,investigators are paying more attention on the pathological process of various kinds of immune complex and complement when they deposit on basement membrane of blood vessel,while little research hit the study point of connected cytokines involved in LCV.Matrix metalloproteinases(MMPs)are zinc-dependent endopeptidases with highly homologous basic structure which can degrade extracellular matrix and maintain its ecological balance.Matrix protease tissue inhibiting factors(TIMPs)can be matched with activated MMPs by 1:1 proportion to form TIMPs-MMPs complex so as to against the degradation process done by MMPs to substrate.The investigations to relationship between vasculitis and MMPs mainly has been concentrated on large or moderate grade vessels vasculitis so far,which revealed the significant role played by MMPs and TIMPs in the development of large or moderate grade vessels vasculitis.To the opposite,studies concerning small vessels especially the relationship between LCV and MMPs in the evolution of small vasculitis is comparatively rare.Among them,one finding is that both MMP-9(gelatinase B)and MMP-3(stromelysins-1)can degrade the specific element of basement membrane of blood vessel,that is ? collagen.In all the four types of TIMP that have been revealed,TIMP-1 is found to be capable of competing all kinds of MMPs,and show high potency to MMP-9 and MMP-3.From histological perspective,LCV may present infiltration of a lot kind of inflammation cells,vascular wall damage and fibrinoid necrosis,karyorrhexis and so on.Based on this,we speculate that MMP-9?MMP-3?TIMP-1 involve in the progression of LCV.IL-37 is one of the IL-1 family members found in recent years.As a new kind of anti-inflammation cytokines,it plays a role in immune response and suppression of a number of pro-inflammatory cytokines.It has been proved that IL-37 acts as a defensive guard in a series of diseases such as systemic lupus erythematosus,rheumatoid arthritis,colitis induced by chemical and liver damage and so on.What's more,it is drawing more and more attention in lots of other dermatological diseases,for example,psoriasis,atopic dermatitis and this like.Nevertheless,the research area concerning relationship between IL-37 and vasculitis is still out of reach.?Object? Patients with LCV were reucrited as the subjects.in present work.different biological sample such as serum,urine and skin lesion were collected from the patients at acute stage of LCV,the expression level of MMP-9?MMP-3?TIMP-1 and IL-37 in different corresponding sample were detected,and realationship were explored among the value and feasibility of the indicators mentioned above in both diagnosis and follow up of LCVwere also discussed.?Method? 1.Subjects: Tissue pathology paraffin blocks from patients diagnosed as LCV ever before and that from LCV patients at present were included.The patients with LCV including allergic purpura(HSP),allergic vasculitis(ACV),urticarial vasculitis(UV)were collected in the department of out or in-patients from March 2016 to January 2017 in The Guangzhou Institute of Dermatology according to the criteria made in 2012 in the America Chapel Hill meeting.Of 30 LCV patients included in the study group,with an average age of 35.0±3.45 years old,and 13 were male,17 were female.Of 10 subjects in the control group,4 were male,6 were female,and with the average age of 38.4±3.68 years old.(2)Sixty-two Paraffin blocks clinically and pathologically confirmed LCVwhich were preserved in Pathology department of our hospital were selected from January 2011 to September 2016,among which 24 were allergic vasculitis(ACV)patients,18 allergic purpura(HSP)patients,20 urticarial vasculitis(UV)patients.2.Sample collecting: Both serum and urine were collected for laboratory analyze at the first visit and subsequent visit 3 weeks after treatment respectively,and biopsy was performed at first visit too.3.Detection of indicators:(1)The concentration of MMP-9,MMP-3,TIMP-1 and IL-37 levels of serum and urine were determined respectively by enzyme-linked immunosorbent assay(ELISA);(2)Expression of MMP-9,MMP-3,TIMP-1 and IL-37 of paraffin specimens were detected by immunohistochemistry;(3)The m RNA transcription of MMP-9,MMP-3,TIMP-1 and IL-37 were assessed by the reverse transcription real-time quantitative PCR;4.Statistical analysis: Using SPSS 16.0 software for statistical analysis,comparing the difference between the LCV group and normal control group and correlation analysis was carried out on the different indicators.P < 0.05 as statistically significant??Result? ?.Syudy on both serum expression level and relationship of MMPs?MMPs/TIMP and IL-37 for LCV patients at acute stage1.Results of MMPs?MMPs/TIMP?IL-37 before treatment:(1)MMP-9 in the serum levels of LCV group(1580.48±206.51)ng/ml was is significantly higher than that of control group(492.19±107.96)ng/ml(P < 0.01);MMP-9/TIMP-1 of LCV group 16.71±2.76 was is significantly higher than that of control group 4.54±1.14(P < 0.01);IL-37 in serum levels of LCV group(214.79±16.39)ng/ml was significantly lower than that of control group(417.82±40.91)ng/ml(P < 0.01).(2)There was no significant difference for MMP-3,TIMP-1,and MMP-3/TIMP-1 between study group and control group(P > 0.05);(3)Moderate positively correlated was found by spearman correlation analysis between TIMP-1 and MMP-9(r=0.555,P<0.01);and so as TIMP-1 and MMP-3 too(r=0.547,P<0.01).On the contrary,slightly negatively correlated was shown between MMP-3 and IL-37(r=-0.397,P<0.05),and so as IL-37 and MMP-9/TIMP-1 too(r=-0.353,P<0.05).2.Results of MMPs?MMPs/TIMP?IL-37 after treatment: Serum MMP-9 concertntration declined by 60% when compared to its original level(957.36±283.71ng/ml vs 381.24±98.16)ng/ml;and similar for TIMP-1 by 52%(228.33±92.4 ng/ml vs 108.56±20.07ng/ml),MMP-9/TIMP-1 by 58%(11.35±8.31 vs 4.79±2.54);on the other hand,IL-37 increased approximately by 155% when compared to its original level(290.54±35.70 ng/ml vs 741.91±191.30ng/ml).Yet no significant difference was observed in MMP-3,and MMP-3/TIMP-1.?.Syudy on both urine expression level and relationship of MMPs ?MMPs/TIMP and IL-37 for LCV patients at acute stage1.Results of MMPs?MMPs/TIMP?IL-37 before treatment: MMP-9 in the urine levels of LCV group(587.08±131.04)ng/ml was significantly higher than that of control group(159.44±51.10)ng/ml(P < 0.05).But no significant difference was observed for MMP-3,TIMP-1,MMP-9/TIMP-1 and MMP-3/TIMP-1when compare d to control group(P>0.05).2.Results of MMPs ? MMPs/TIMP ? IL-37 after treatment:Urine MMP-9 concentration declined by 75% when compared to original level(1106.20±423.70ng/ml vs 81.21±88.43ng/ml)ng/ml,and similar for MMP-9/TIMP-1 declined by 68%(56.50±30.58 vs 18.47±9.25).Yet no significant difference was observed in MMP-3,MMP-3/TIMP-1,TIMP-1and IL-37(P>0.05).?.The comparison of tissue expression intensity of MMPs?TIMP-1?IL-37 in lesion of LCV at acte stage:1.Expression of MMPs and TIMP-1: MMP-9 mainly expresses in keratinocyte,lymphocyte,Neutrophils and cytoplasm of endothelial cells.In this study,it is found that the expression level in LCV patients is significantly higher than control group(0.069±0.001 vs 0.013±0.001,P<0.01).MMP-3 mainly expresses in a few keratinocytes and exceptional endothelial cells,and expression level was not significantly differed between LCV group and control group(P>0.05).TIMP-1 generally expresses in a small amount of basal layer cells in epidermis and lymphocytes in dermis.No significant difference was shown for TIMP-1 between LCV and control group.2.MMPs/TIMP-1: MMP-9/TIMP-1 of LCV group is obviously higher than control group(4.83±0.30 vs 0.14±0.03,P < 0.01),whereas the comparison of MMP-3/TIMP-1 shows no significant difference(0.09±0.01 vs 0.08±0.02,P>0.05).3.IL-37: IL-37 usually expresses in part of keratinocytes,cytoplasm of endothelial cells in dermis and scattered collagen tissue.The expression levels in LCV patients are significantly lower than control group(0.034±0.002 vs 0.096±0.004,P<0.01).?.Comparison of protein expression intensity in immunohistochemical array among different subtypes of LCV.1.The expression intensity of MMP-9 in ACV group,UV group and HSP group were all significantly higher than that of control group(P < 0.01),no significant difference was found among ACV,UV and HSP group(P > 0.05).Moreover,no significant difference was found in the expression of MMP-3 and TIMP-1 between all the subtype groups(P > 0.05).2.IL-37 expression in ACV group,UV group and HSP group were lower than that of control group(P < 0.01),and the expression intensity for both UV and HSP group were lower than that in ACV group(P < 0.01),but the UV group showed no significant difference with HSP group(P > 0.05).?.Comparison of m RNA transcription of MMPs?TIMP-1?IL-37 in LCV patients at acute stage.1.The m RNA transcription level of MMP-9 was dramatically higher in the LCV group than that in the control group(3.10±0.63 VS 0.84±0.28,P<0.01),while The transcription level of IL-37 m RNA was significantly lower in LCV group than that in control group(29.53±4.80 VS 128.31±28.9,P<0.01).2.The m RNA transcription level of MMP-3 and TIMP-1 in LCV group had no significant difference when compared to that in control group(0.32±0.14 VS 0.25±0.14,2.42±1.37 VS 2.07±1.15,P>0.05).?Conclusion? 1.MMP-9,IL-37 may be involved in the occurrence of LCV and disease changes.2.MMP-9 and the radio of MMP-9/TIMP-1 may be positively correlated with the severity of LCV.3.The expression of MMP-9 in the urine of LCV patients was significantly increased,and urine can be used as a noninvasive index to judge the prognosis of LCV.4.The level of IL-37 in LCV patients may be negatively correlated with LCV disease activity.5.MMP-9 m RNA transcription level in patients with LCV was basically the same as that of serum MMP-9,which was higher than that in control group,while IL-37 was lower than that in control group. |
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