【作者】 董海玲；

【导师】 郑凤兰； 高顺强；

【作者基本信息】 河北医科大学 ， 皮肤病与性病学， 2008， 硕士

【摘要】 目的:玫瑰糠疹(Pityriasia rosea, PR)是一种临床较为常见的炎症性皮肤病,皮损为大小不等的圆形或椭圆形的玫瑰色斑疹,其表面附有糠状鳞屑,多发生在躯干及四肢近端。病因及发病机制尚不清楚,涉及病毒感染、变态反应、自身免疫、遗传性过敏症等多种学说。白三烯B4(leukotriene B4, LTB4)在变态反应、炎症、和免疫系统中发挥着重要作用。LTB4是肥大细胞、嗜碱性粒细胞激活后立即合成的介质,通过脱颗粒而向细胞外释放,作用于其它细胞和组织而参与变态反应,在I型变态反应速发期,可以与白介素-4 (interleukin-4, IL-4)共同增加淋巴细胞免疫球蛋白E (Immunoglobin E,IgE)的产生,在变态反应的迟发相中LTB4又是浸润得炎症细胞:巨噬细胞、中性粒细胞合成分泌的介质,且是I型变态反应迟发相所释放的最经典的炎症介质,在I型变态反应中可引起广泛的炎症损伤。变态反应对机体而言是防御机制的免疫反应,但却会对机体产生伤害性作用。5-羟色胺(5-hydroxytryptamine, serotonin, 5-HT)是炎性介质之一,当体内发生体液性或细胞性免疫反应时,可引起介质的激活或释放,并作用于效应细胞的受体,造成组织损伤,产生炎症反应, 5-羟色胺是I型变态反应迟发相所释放的一种血管活性胺类物质,具有与组胺和过敏性慢反应物质一样收缩平滑肌和增加血管通透性的作用,可引起皮肤红斑、水肿、发绀、出血、瘙痒等炎症反应。白介素-1β(interleukin,IL-1β)是一种主要起免疫调节作用的激素样肽类物质,即炎症前期细胞因子,主要由活化的单核-吞噬细胞产生,释放IL-1β是炎症急性期最基本和最重要的免疫反应现象。本实验通过酶联免疫吸附试验法检测PR患者血清LTB4水平,免疫组织化学技术检测PR患者皮损5-HT、IL-1β表达情况来阐述其在PR发病机制中的作用,并探讨玫瑰糠疹与变态反应的关系。材料与方法:血清标本均来源于2006年12月~2007年6月到河北医科大学第四附属医院皮肤科就诊的临床诊断为玫瑰糠疹的患者,共30例,其中男性16例,女性14例,年龄15~40岁,平均(24.58±6.19)岁,病程2天~2个月,平均(17.50±14.46)天,并且随机选取10例健康志愿者作为正常对照,对照组在年龄、性别上均与玫瑰糠疹患者组相匹配。所有受检者均无其他LTB4异常的疾病(如呼吸系统疾病、其他过敏性疾病、其他炎症性疾病、自身免疫性疾病等),一个月内无系统用药史(以除外药疹)及冶游史(以除外二期梅毒疹),同时未使用过抗组胺药、抗白三烯药及免疫抑制剂。皮损标本均来源于2003年3月~2007年5月到河北医科大学第四附属医院皮肤科就诊的玫瑰糠疹患者30例,均经临床和组织病理确诊为玫瑰糠疹,其中男性14例,女性16例,年龄10~42岁,平均(29.73±3.85)岁,病程5~40天,平均(19.89±3.88)天。所有患者均已排除患有其他系统性疾病,且3个月内未接受任何免疫、抗炎及抗过敏治疗。选取河北医科大学第四附属医院外科10例手术患者的健康皮肤作为正常对照,对照组在年龄、性别及取材部位上均与玫瑰糠疹患者相匹配。应用酶联免疫吸附法(Enzyme linked immunosorbent assay,ELISA)检测血清LTB4水平,结果采用两独立样本比较的t检验。不同性别的结果亦采用两独立样本比较t的检验,不同年龄及瘙痒程度的结果采用单因素方差分析。取患者皮损区组织做常规组织病理分析,应用免疫组织化学SP法检测5-HT及IL-1β的表达情况,结果采用两等级资料比较的Mann-whiteney U秩和检验以及Spearman等级相关分析。应用SPSS11.5统计软件分析试验结果。结果1 ELISA结果1.1玫瑰糠疹组血清LTB4含量均值1.8632±0.1605,与正常对照组均值1.4484±0.1044比较,经统计学分析,两组间有差异(P﹤0.05)。1.2不同性别、年龄及瘙痒程度玫瑰糠疹患者血清LTB4含量的均值,经统计学处理,各组间均无差异(P﹥0.05)。2免疫组化结果阳性染色为棕黄色颗粒。2.1 5-HT在玫瑰糠疹皮损表皮中呈阳性表达,主要表达于棘细胞层及颗粒层的细胞浆,30例玫瑰糠疹皮损中,强阳性0例,阳性5例,弱阳性17例,阴性8例,正常表皮不表达,两者差异有显著性(P﹤0.01)。2.2 IL-1β为阳性细胞浆及胞膜着色,表皮全层广泛阳性着色,30例玫瑰糠疹皮损中,强阳性3例,阳性11例,弱阳性14例,阴性2例,与正常对照组比较明显增高,经统计学分析有差异(P﹤0.05)。2.3玫瑰糠疹患者皮损中5-HT和IL-1β的表达,经统计学分析,两者有相关性(P﹤0.01)。结论1在本实验中,玫瑰糠疹患者组血清LTB4水平升高,提示LTB4可能参与了PR的发生或发展,它的作用可能是通过趋化白细胞向炎症部位聚集并诱导白细胞脱颗粒和产生超氧离子直接造成组织损伤,使白细胞黏附于血管内皮细胞,引起白细胞依赖性的血管通透性增加,与其它致炎因素共同导致了PR的发生发展。LTB4作为I型变态反应迟发相所释放的最经典的炎症介质,提示I型变态反应可能参与了PR的发病过程。2皮损中5-HT的过度表达,提示PR的发病机制中可能有5-HT的参与。5-HT可能是作为I型变态反应迟发相所释放的血管活性胺类物质,使皮肤毛细血管扩张,通透性增高,引起皮肤红斑等炎症反应,参与了PR患者皮损的发生,亦可能是通过5-HT的其他功能参与了PR皮损的形成和发展。3 IL-1β在玫瑰糠疹皮损中的过度表达,提示其可能作为一种重要的炎症介质,参与了PR的发病过程,机理可能是促使血管内皮细胞表达内皮细胞-白细胞黏附分子-1和细胞间黏附分子-1,从而增强白细胞与内皮细胞的黏附作用,促进炎细胞渗出与迁移,激活炎细胞,与其它因素共同造成了机体免疫损伤。4皮损中5-HT和IL-1β的表达,经统计学分析两者有相关性提示其作为致炎因子,可能互相促进共同参与了玫瑰糠疹的发病过程。更多还原

【Abstract】 Objective: Pityriasis rosea (PR) is an inflammation skin disease, which is common in the Department of Dermatology. The lesions tend to be distributed on the trunk and extremities. The primary macules are yellow-red ,oval or circular and covered with small, thin scales like a collar. The etiology and pathogenesis of PR remain unkown. Now many people consider that it is related with virus infection, hypersensitivity reaction, autoimmunity, genetic anaphylaxis and so on. Leukotrinene B4(leukotriene B4,LTB4)plays an important role in the hypersensitivity,inflammation,and immune system.LTB4 is synthesised immediately by mast cells and basophilic leukocyte degranulate when they are activited and realeased through degranulated way.LTB4 partcipated hypersensitivity reaction through other cells and issues.LTB4 enhances lymphocytes to produce IgE with IL-4 together in typeⅠhypersensitivity early stage, and it is factor synthesized by inflammation cells such as macrophages and neutrophil infiltrated in typeⅠhypersensitivity late stage, it is the classic inflammation factor realeased in typeⅠhypersensitivity late stage. LTB4 can cause wide inflammation in typeⅠhypersensitivity. Hypersensitivity reaction is a protected immune reaction for human body, but witch can hurt the body sometimes.5-hydroxytryptamine is one of inflammatory mediators.When fluids or cells immune is responsed then the meditors are activated or realeased witch hurt the issues through their receptor cell and cause inflammation.5- hydroxytryptamine is one of vasoactive amines material realeased in late stage on typeⅠhypersensitivity ,it can cause smooth muscle shrinkages and increased vascular`s permeability just like histamine and allergic slow-reacting substance. 5-hydroxytryptamine may cause smooth muscle shrinkages, capillaries expansion and enhance the permeability then cause skin erythema, edema, cyanosis, bleeding, itching, and other inflammatory response. IL-1βis an hormone-like peptide substance and its main function is to regulate immune. IL-1βis mainly generated by the activated monocytes– phagocytes. Reased IL-1βis the primary and the most important immune reponse phenomenon. This study detected the serum concentration of IL-1βby ELISA(Enzyme linked immunosorbent assay) and observe the expressions of 5-hydroxytryptamine and IL-1βin PR lesions to explain their function in the pathogenesis of PR. At the same time to explore the relation between PR and hypersensitivity.Materials and Methods: Thirty PR serum samples includigng 14 males and 16 females were collected from PR patients diagnosed by clinic of the dermatological department of the Forth Affiliated Hospital of Hebei Medical University during Dec,2006 to June,2007. Age ranged from 15 to 40 years(mean 24.58±6.19); Course from 2 days to 2 months,(mean 17.50±14.46). And random select 10 out–hospital healthy volunteers as controls. There are no significant differences in sexes and ages between controls and patients. All patients and volunteers have no LTB4 novel diseases(for example respiratory diseases, other allergic diseases, inflammatory diseases, autoimmune diseases), and had on systematic mediation history as well as swim history (to exclusion durg eruption and PR-type secondary syphilis) within one month, had no taken immunodepressants and antihistamine at the same time.Thirty lesion case diagnosed as PR clinically and histopathologically were collected from the dermatological department of the Forth Affiliate Hospital of Hebei Medical University during Mar. 2003 to May.2007, who had no other systemic disease or other skin diseases,and had not received any treatment since they were selected. These cases included 14 males and 16 females, age arranged from 10 to 42 years (mean29.73±3.85), course arranged from 5 to 40 days (mean19.89±3.88). The ten normal controls were healthy volunteers in the hospital, and had no significant difference in sex, age and position between patients and controls.The sera concentration of LTB4 were measured by enzyme linked immunosorbent assay (ELISA), the result were analyzed by 2-independent sample compared T test. The result of gender was analyzed by2-independent sample compared T test and the results of different age and pruritus degree were analyzed by one-factor analysis of variance. The lesion samples were investigated with histopathological method.The levels of 5-HT and IL-1βexpressions were measured by immunohistochemical method. HE-staining was used in histopathology and histostain-SP in immuno-histochemistry. The results were analyzed by Mann-whitney U rand sum test and Spearman’s rank correlation test.Results1 ELISA results1.1 The mean concentrations of sera LTB4 of PR patients and normal controls were1.8632±0.1605, and1.4484±0.1044, compared with each other, there was statistal significance (p<0.05).1.2 The mean concentrations of sera LTB4 of PR patients, which compared from gender, age phases and pruritus degrees, there were no statistal significance (p>0.05).2 Immunohistochemistry results Positive staining was submitted Buffy grains.2.1 5-HT stained on cytoplasm of stratum spinosum and stratum granulosum cell. Among these positive cells, 0 cases (+++), 5cases (++), 17 cases (+), 8 case (-).The level of its expression was markedly increased comparing with the controls (p<0.01).3.2 IL-1βstained on cell membranes and cytoplasm of full epidermis cell . Among these positive cells, 3 cases (+++), 11cases (++), 14 cases (+), 2 case (-).The level of its expression was markedly increased comparing with the controls (p<0.05).3.3 There was significant correlation between the expression degree of HSP-70 and TNF-α(p<0.01).Conclusion1 In this study, the concentration of serum LTB4 of PR patients is significantly increased, this implies LTB4 participants the development of PR. The mechanism may be LTB4 chemokines leukocyte aggregated to inflammation site and induces leukocyte to degranulate and prduce superoxide ions to hurt issue directly, then make leukocyte adhesion to endothelial cells, increase interleukin-dependent vascular permeability and induce the development of PR with other factors. LTB4 be the classic factor in typeⅠhypersensitivity late stage implies typeⅠhypersensitivity maybe participate the development of PR.2 The expression of 5-hydroxytryptamine in the lesions is stronger than that of controls , it explains that 5-serotoin may be take part in the development of PR.( It may be vasoactive amines material in typeⅠhypersensitivity late stage cause skin`s capillaries expansion ,permeability enhanced lead the skin emergy erythea and other inflammation reaction to participant the produce of PR It is perhaps that 5-HT takes part in the development of PR by it`s other fuction.3 The expression of IL-1βin the lesions is stronger than that of controls implies that it be of an important inflammation factors take part in the development of PR.The mechanism maybe IL-1βmake vascular endothelial cells expression endothelial cells-leukocyte adhesion molecule-1 and intercellular adhesion molecule-1,then enhancing the adhesion between interleukins, promoting exudation and migration of inflammatory cells and activating inflammation ,cause the immune hurt to the body with other factors.4 There was significant correlation between the expression degree of 5-HT and IL-1β,that implies they be of inflammation factors promting eatch other to take part in the development of PR.更多还原