Le psoriasis est une maladie cutanée grave pour laquelle il n’existe encore aucun traitement curatif. Des modèles de peau psoriasique in vitro sont alors nécessaires afin de tester de nouveaux traitements. Cette étude visait, en premier lieu, à optimiser la production de substituts cutanés psoriasiques par la méthode d’auto-assemblage, pour ensuite les utiliser afin d’évaluer l’efficacité d’une nouvelle molécule potentiellement antipsoriasique, la tBEU. Dans un premier temps, des substituts sains et psoriasiques ont été produits par la méthode d’auto-assemblage partiellement modifiée en utilisant des plaques 6 puits et 12 puits. Ces expériences ont permis de démontrer que les plaques 6 puits étaient plus efficaces pour la production de substituts reproductibles et représentatifs d’une peau psoriasique in vivo. Ensuite, ces substituts ont été traités avec la tBEU, démontrant que cette molécule diminue la prolifération des kératinocytes et améliore leur différenciation. Ainsi, la tBEU pourrait éventuellement être utilisée comme un traitement efficace du psoriasis. / Psoriasis is a serious cutaneous disease for which there is currently no cure. In vitro psoriatic skin models could become new tools to assess the potency of new drugs during the development of new therapies. First, this study aimed to optimize the production of psoriatic skin substitutes by the self-assembly method, and then use them to assess the effectiveness of a new potentially antipsoriatic molecule, tBEU. Initially, healthy and psoriatic substitutes have been produced by the self-assembly method partially modified, using 6-well and 12-well plates. These experiments demonstrated that the 6-well plates were more effective for the production of reproducible substitutes and representative of psoriatic skin in vivo than the 12-well plates. Then, these substitutes were treated with tBEU, showing that this molecule decreases significantly keratinocytes proliferation and improves their differentiation. Thus, tBEU may eventually be used as an effective topical treatment for psoriasis.

Le psoriasis se caractérise par la présence d’infiltrats leucocytaires inflammatoires au niveau du derme et de l’épiderme cutané humain, menant à des désordres cutanés considérables : hyperprolifération des cellules épidermiques basales, différenciation altérée des kératinocytes et sécrétion accrue de médiateurs pro-inflammatoires dans le microenvironnement local. Cette dysfonction immuno-épithéliale menant au développement de lésions blanchâtres, agit tel un cercle vicieux incontrôlable, stimulant réciproquement les kératinocytes et les cellules immunitaires. Dans cette optique, mes travaux de doctorat ont visé à : (1) générer un microenvironnement immunocompétent par l’intégration de cellules T pré-activées dans un modèle de peau saine, non-lésionnelle et lésionnelle, produit selon la méthode d’auto-assemblage afin (2) d’étudier le rôle de cette composante importante sur les caractéristiques-clés associées au psoriasis. De surcroit, nous avions pour autre objectif (3) d’analyser l’impact de macrophages dérivés de monocytes dans un modèle tridimensionnel de peau saine afin (4) de déterminer l’influence de ces cellules sur le remodelage matriciel et la différenciation épidermique à l’état d’équilibre pour, finalement, (5) analyser l’empreinte du microenvironnement sur leur polarisation in vitro. Nos données mettent en lumière le caractère déterminant de la composante immunitaire, lorsqu’associée aux cellules cutanées pathologiques, dans le développement des lésions psoriasiques, puisque la présence de cellules T, dans un modèle de peau saine, n’induit pas l’ensemble des caractéristiques histopathologiques associées à cette dermatose. D’autre part, ces modèles de peau immunocompétente répondent à un traitement de choix pour le psoriasis, le méthotrexate, qui inhibe la prolifération cellulaire et atténue l’inflammation. Ces nouveaux modèles organotypiques serviront d’outils précliniques performants pour le criblage d’actifs visant à traiter certaines pathologies d’origine auto-immune inflammatoire chronique, dans lesquelles le système immunitaire joue un rôle crucial, à l’exemple du psoriasis. Par ailleurs, l’intégration de macrophages dérivés de monocytes dans un modèle de peau normale, affecte l’état de différenciation des kératinocytes épidermiques tout en réduisant l’état inflammatoire basal. Ces macrophages induisent, en outre, un remodelage matriciel tissulaire important modifiant de ce fait le microenvironnement avoisinant. Finalement, ce nouveau modèle tridimensionnel nous a permis, d’une part, de mimer davantage la structure cutanée physiologique et, d’autre part, de mieux comprendre l’influence de ces cellules sur le phénotype cutané à l’état d’équilibre. Bien que d’autres études soient nécessaires avant l’utilisation de ces modèles en phase préclinique, ces travaux constituent une avancée majeure dans le développement de modèles produits par génie tissulaire, plus complexes et plus pertinents d’un point de vue physiologique. / Psoriasis is characterized by the presence of inflammatory leukocyte infiltrates in the human skin dermis and epidermis, leading to important skin disorders: hyperproliferation of basal epidermal cells, altered differentiation of keratinocytes, and increased secretion of proinflammatory mediators in the local microenvironment. This immuno-epithelial dysfunction leading to the development of whitish lesions acts as an uncontrollable vicious circle, stimulating the keratinocytes and the immune cells. With this in mind, my thesis work aimed to: (1) generate an immunocompetent microenvironment by integrating pre-activated T cells into a healthy, non-lesional and lesional skin model, produced using the self-assembly method (2) to study the role of this essential component on key features associated with psoriasis. In addition, our other objective was (3) to analyze the impact of monocyte-derived macrophages in a three-dimensional model of healthy skin (4) to determine the influence of these cells on tissue remodeling and epidermal differentiation at steady state, (5) in order to finally analyze the microenvironment effect on their in vitro polarization. Our data highlight the essential feature of the immune component, when associated with pathological skin cells, in the development of psoriatic lesions, since the presence of T cells, in a healthy skin model, does not induce all of the histopathological key features, associated with the dermatosis. On the other hand, these models of immunocompetent skin respond to a treatment of choice for psoriasis, methotrexate, which inhibits cell proliferation and reduces inflammation. These new organotypic models will serve as powerful preclinical tools for the screening of active agents to treat some chronic inflammatory pathologies of autoimmune origin, in which the immune system plays a crucial role, like psoriasis. Moreover, the integration of monocytederived macrophages into a normal skin model affects the differentiation state of epidermal keratinocytes while reducing the basal inflammatory state. These macrophages induce, in addition, a significant matrix remodeling thereby modifying the surrounding microenvironment. Finally, this new three-dimensional model allowed us, on the one hand, to further mimic the physiological cutaneous structure and, on the other hand, to better understand the influence of these cells on the skin phenotype at steady state. Although further studies are needed prior to the use of these preclinical models, this work represents a major breakthrough in the development of more complex tissue-engineered models that are physiologically relevant.

Die Psoriasis ist mit einer Prävalenz von 1-3% eine der häufigsten chronisch-entzündlichen Erkrankungen. Histopathologisch ist sie gekennzeichnet durch epidermale Hyperkeratose und Akanthose, gesteigerte Angiogenese sowie ein gemischtzelliges leukozytäres Infiltrat. Aus aktueller Sicht wird sie als eine komplexe primär T-Zell vermittelte Autoimmunerkrankung mit genetischer Prädisposition verstanden. Psoriasis kann mit kardiovaskulären, metabolischen und psychiatrischen Erkrankungen assoziiert sein (Komorbidität). Hier wurde erstmals die Wirkung des inositoylierten Plättchen-aktivierenden Faktors (Ino-C2-PAF), eines synthetischen Alkylphospholipids mit geringer Toxizität, in experimentellen Modellen chronisch-entzündlicher (Haut)-Erkrankungen untersucht. Die Wirkung beruht auf Zellmembran-Interaktionen, wodurch der Cholesterol- und Phospholipidmetabolismus verändert und zelluläre Signalkaskaden der Proliferation, Apoptose und Motilität beeinträchtigt werden. Ino-C2- PAF reguliert außerdem entzündungsrelevante Proteine herab. In vitro lag der Schwerpunkt auf der Wirkung von Ino-C2-PAF auf Endothelzellen und Leukozyten. Ino-C2-PAF hemmte die Proliferation humaner Endothelzellen moderat, steigerte allerdings die Apoptose TNFα-stimulierter Endothelzellen sehr deutlich. Diese Wirkung war begleitet von einer Reduktion der durch TNFα stimulierbaren endothelialen Adhäsionsmoleküle VCAM-1, ICAM-1 und E-Selektin sowie der lymphozytären Adhäsionsmoleküle CD49d, CD11a, CD62L und CLA. Funktionell führte dies zu einer signifikanten Abnahme dynamischer Interaktionen (Rollen und feste Adhäsion) von Leukozyten und aktivierten Endothelzellen in Flusskammerexperimenten. Die anti-entzündliche Wirkung von Ino-C2-PAF wurde in zwei komplementären Modellen in vivo bestätigt. Sowohl in K5.hTFGß-transgenen als auch in JunB/c-Jun-defizienten Mäusen bewirkte Ino-C2-PAF eine signifikante Besserung des psoriasiformen Phänotyps sowohl auf makroskopischer als auch auf histopathologischer Ebene. Die Ergebnisse dieser Arbeit legen nahe, dass Ino-C2-PAF oder verwandte Substanzen zur Behandlung entzündlicher Erkrankungen wie der Psoriasis eingesetzt werden könnten.

銀屑病是一種免疫相關的慢性炎症性皮膚病，其發病率約占世界人口的1-3%，而現今仍然缺乏有效安全的根治方法。國內外使用中草藥治療銀屑病取得較好的療效，但目前缺少對其進行系統研究和開發。我們研究小組之前對61種常用治療銀屑病中藥進行篩選, 發現中藥茜草根和南蛇藤的乙醇提取物具有強大的抑制表皮細胞增生的作用，本博士研究課題的目的是確定新的安全有效的用于治療銀屑病的中藥化學成分, 並闡明其作用機制。 / 本研究篩選了28種存在于這兩種中藥中的化學單體成分，采用體外培養永生化的人類皮膚良性角質形成細胞株HaCaT, 應用MTT法, 繪制細胞生長曲線，獲得抑制50%細胞生長所需藥物濃度(IC50)。實驗結果發現1-羟基-3-甲基蒽醌（HMA）， 1，4-二氨基-2，3-（2-苯氧基乙氧基）蒽醌 （DBA）和南蛇藤表現了強大的抗表細胞生長作用，其48小時培養後的IC50分別爲17.9，15.8，1.1 μM. 值得一提的是這些化合物對正常人表皮角質細胞HEK和人類成纖維細胞Hs68只有相對輕微細胞毒性。 / 隨後進行的機理研究，通過熒光染色，DNA凝膠電泳，細胞周期檢測，流式細胞計檢測及Western blot 分析結果表明， HMA和南蛇藤素是通過誘導細胞凋亡作用抑制HaCaT細胞生長。其中南蛇藤素通過線粒體凋亡和死亡受體介導的兩種通路誘導細胞凋亡， 其誘導細胞凋亡作用與其抑制核因子-κB在HaCaT細胞中的表達和活化有關。 / 另一方面，DBA 抑制人體表皮角質細胞生長的作用機理在于其對角質細胞終末分化的誘導作用。DBA與HaCaT和HEK細胞共同培養96小時後，能顯著促進細胞角質化外膜形成，同時上調角蛋白K1/10，人體套膜蛋白，轉谷氨酰胺酶-1表達和下調角蛋白K5/14表達。而利用小鼠尾部鱗片表皮模型對HMA的外用制劑進行測試，結果顯示HMA誘導角質細胞終末分化能力較弱。 / 總而言之，本研究課題從兩種中藥中成功發現三個具有較強的抗銀屑病活性的化學單體成分，這些來自中藥的天然産物具有很好的開發成新的銀屑病治療外用制劑的應用前景。 / Psoriasis is an immunologically-mediated chronic inflammatory disease of the skin and joints affecting approximately 1-3% of the world’s population. Traditionally, Chinese medicine has been extensively used both inside and outside China for treating psoriasis with promising clinical results. Based on the promising findings in our previous screening project on 61 psoriasis-treating Chinese medicines which showed the root of Rubia cordifolia L. (Rubiae Radix) to have potent anti-psoriatic action, the present study aimed to identify active anti-psoriatic chemical constituents derived from Rubiae Radix and another Chinese herb namely Celastrus orbiculatus Thunb. and to elucidate the underlying mechanisms of action. / Microplate MTT assay was performed to evaluate the anti-proliferative actions of 28 selected Rubiae Radix-derived anthraquinones and other chemical ingredients on cultured HaCaT keratinocytes. Among them, 1-hydroxy-3-methyl-anthraquinone (HMA) and 1,4-diamino-2,3-bis(2-phenoxyethoxy)anthraquinone (DBA), as well as celastrol, a Celastrus orbiculatus-derived triterpene, were found to possess significant anti-proliferative action on HaCaT cells, with IC₅₀ value of 17.9, 15.8 and 1.1 μM, respectively. All DBA, HMA and celastrol showed only mild to moderate toxic effects on normal human keratinocyte HEK cells and human fibroblast Hs68 cells. / Mechanistically, celastrol and HMA was found to induce apoptosis in a dose-dependent manner in HaCaT cells as characterized by DNA fragmentation, phosphatidyl-serine externalization and activation of caspase 3. Further studies by flow cytometric and western blot analyses demonstrated that the celastrol-induced apoptosis on HaCaT cells was associated with the inhibition of NF-κB pathway and through caspase-related apoptotic pathway as characterized by activation of caspase proteins, regulation of Bcl-2 family proteins and depolarization of mitochondrial potential. / On the other hand, DBA showed an ability to induce terminal differentiation in cultured human keratinocytes and this capability is believed to be responsible for its growth inhibitory effects. DBA significantly accentuated the cornified envelope formation in HEK and HaCaT keratinocytes together with the augmentation of K1/K10, involucrin and transglutaminase 1 protein levels and decrease of expression of K5/K14 protein in DBA-treated cells. However, the subsequent in vivo study using a mouse tail model showed that HMA did not have significant effects on modulating keratinocyte terminal differentiation. / Taken together, our present PhD project successfully identified DBA, HMA and celastrol to have potent anti-psoriatic action on in vitro models, and the experimental findings render these naturally-occurring chemicals to be promising candidates for further development into anti-psoriatic pharmaceutical agents. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Zhou, Linli. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 213-244). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iii / Publications --- p.v / Acknowledgements --- p.vii / Table of Contents --- p.viii / List of Figures --- p.xvii / List of Tables --- p.xxi / List of Abbreviations --- p.xxii / Chapter Chapter One --- General Introduction / Chapter 1.1 --- Psoriasis --- p.2 / Chapter 1.1.1 --- Structure of skin --- p.2 / Chapter 1.1.2 --- Epidemiology of psoriasis --- p.3 / Chapter 1.1.3 --- Pathogenesis --- p.5 / Chapter 1.1.4 --- Classification --- p.8 / Chapter 1.1.4.1 --- Nonpustular (plaque type) psoriasis --- p.9 / Chapter 1.1.4.2 --- Guttate psoriasis --- p.9 / Chapter 1.1.4.3 --- Pustular psoriasis --- p.9 / Chapter 1.1.4.4 --- Erythrodermic psoriasis --- p.10 / Chapter 1.1.4.5 --- Nail psoriasis --- p.11 / Chapter 1.1.4.6 --- Psoriatic arthritis --- p.11 / Chapter 1.1.5 --- Comorbidities --- p.13 / Chapter 1.2 --- Treatment of Psoriasis --- p.16 / Chapter 1.2.1 --- Conventional treatment for psoriasis --- p.16 / Chapter 1.2.1.1 --- Topical therapy --- p.16 / Chapter 1.2.1.2 --- Phototherapy --- p.19 / Chapter 1.2.1.3 --- Systemic therapy --- p.21 / Chapter 1.2.2 --- Targeted immunotherapy --- p.24 / Chapter 1.2.3 --- Combination, rotational and sequential therapy --- p.25 / Chapter 1.2.4 --- Complementary treatment --- p.26 / Chapter 1.3 --- Traditional Chinese Medicine for Psoriasis --- p.30 / Chapter 1.3.1 --- Prescriptions for psoriasis based on pattern differentiation --- p.30 / Chapter 1.3.2 --- Clinical and experimental study of TCM for psoriasis --- p.34 / Chapter 1.3.3 --- Possible action mechanisms of Chinese herbs for psoriasis --- p.34 / Chapter 1.3.4 --- Previous studies on TCM for psoriasis conducted by our research group --- p.35 / Chapter 1.4 --- Aims and Objectives of the Present Study --- p.38 / Chapter Chapter Two --- Phytochemical and Apoptotic Studies of Rubiae Radix-derived Anthraquinones and Other Related Compounds / Chapter 2.1 --- Introduction --- p.41 / Chapter 2.2 --- Selection and Screening of Rubiae Radix-derived anthraquinones and Other Related Compounds for Anti-proliferative Action on Cultured HaCaT Human Keratinocytes --- p.43 / Chapter 2.2.1 --- Introduction --- p.43 / Chapter 2.2.2 --- Materials and methods --- p.45 / Chapter 2.2.2.1 --- Procurement of Rubiae Radix-derived anthraquiones and other related compounds --- p.45 / Chapter 2.2.2.2 --- Purification of anthraquinones from Rubiae Radix --- p.50 / Chapter 2.2.2.3 --- General cell culture --- p.54 / Chapter 2.2.2.4 --- SRB assay --- p.55 / Chapter 2.2.2.5 --- MTT assay --- p.56 / Chapter 2.2.2.6 --- Assessment of synergistic or antagonistic effects between two active anthraquiones --- p.56 / Chapter 2.2.2.7 --- Statistical analysis --- p.57 / Chapter 2.2.3 --- Results --- p.57 / Chapter 2.2.3.1 --- Anti-proliferative effects of 35 Rubiae Radix fractions on HaCaT cells by SRB assay --- p.57 / Chapter 2.2.3.2 --- Anti-proliferative effects of the 27 anthraquinones and related compounds on HaCaT cells by SRB assay --- p.59 / Chapter 2.2.3.3 --- Confirmation of the anti-proliferative action of 8 active pure compounds using MTT assay --- p.61 / Chapter 2.2.3.4 --- Cytotoxic effects of 1-hydroxy-3-methyl-anthraquinone and1,4-diamino-2,3-bis(2-phenoxyethoxy)anthraquinone on the growth of HEK and Hs68 cells --- p.64 / Chapter 2.2.3.5 --- Drug interactions between different active anthraquinones --- p.67 / Chapter 2.2.4 --- Discussion --- p.69 / Chapter 2.3 --- Investigations of the Apoptotic Effects of DBA and HMA on HaCaT cells --- p.71 / Chapter 2.3.1 --- Introduction --- p.71 / Chapter 2.3.2 --- Materials and methods --- p.76 / Chapter 2.3.2.1 --- Chemicals --- p.76 / Chapter 2.3.2.2 --- General cell culture methods --- p.76 / Chapter 2.3.2.3 --- Cell cycle analysis with PI staining --- p.76 / Chapter 2.3.2.4 --- Hoechst fluorescence staining for morphological evaluation --- p.77 / Chapter 2.3.2.5 --- DNA fragmentation assay --- p.77 / Chapter 2.3.2.6 --- Detection of apoptosis by flow cytometry --- p.78 / Chapter 2.3.2.7 --- Prepare cytosol fraction of HaCaT cells --- p.79 / Chapter 2.3.2.8 --- Western blot analysis --- p.79 / Chapter 2.3.2.9 --- Statistical analysis --- p.80 / Chapter 2.3.3 --- Results --- p.76 / Chapter 2.3.3.1 --- Action of DBA and HMA on cell cycle progression --- p.80 / Chapter 2.3.3.2 --- Alteration of cellular morphology --- p.84 / Chapter 2.3.3.3 --- Detection of DNA fragmentation --- p.86 / Chapter 2.3.3.4 --- Quantitative analysis of apoptotic cells by annexin V-PI staining --- p.88 / Chapter 2.3.3.5 --- Activation of procaspase-3 and release of cytochrome c protein --- p.91 / Chapter 2.3.4 --- Discussion --- p.94 / Chapter 2.4 --- General Discussion --- p.97 / Chapter Chapter Three --- Effects of Rubiae Radix and Its-derived Anthraquinones on Keratinocyte Terminal Differentiation / Chapter 3.1 --- Introduction --- p.100 / Chapter 3.2 --- Materials and Methods --- p.105 / Chapter 3.2.1 --- Chemicals --- p.105 / Chapter 3.2.2 --- General cell culture --- p.105 / Chapter 3.2.3 --- Cornified envelope (CE) formation assay --- p.106 / Chapter 3.2.4 --- Western blot analysis --- p.107 / Chapter 3.2.4 --- Statistical analysis --- p.107 / Chapter 3.3 --- Results --- p.108 / Chapter 3.3.1 --- EA fraction of Rubiae Radix, DBA and HMA stimulates CE formation --- p.108 / Chapter 3.3.2 --- EA fraction of Rubiae Radix, DBA and HMA regulated TG1 expression and involucrin production in cultured human keratinocytes --- p.112 / Chapter 3.3.3 --- Regulation of cytokeratins by EA fraction of Rubiae Radix, DBA and HMA --- p.118 / Chapter 3.4 --- Discussion --- p.128 / Chapter Chapter Four --- Anti-psoriatic Action of Celastrol from Celastrus orbiculatus / Chapter 4.1 --- Introduction --- p.136 / Chapter 4.2 --- Anti-proliferative Action of Celastrol on Cultured Human Keratinocytes and Other Cell Types --- p.138 / Chapter 4.2.1 --- Introduction --- p.138 / Chapter 4.2.2 --- Materials and methods / Chapter 4.2.2.1 --- Chemicals --- p.138 / Chapter 4.2.2.2 --- General cell culture --- p.139 / Chapter 4.2.2.3 --- MTT assay --- p.139 / Chapter 4.2.2.4 --- Statistical analysis --- p.139 / Chapter 4.2.3 --- Results --- p.142 / Chapter 4.2.3.1 --- Anti-proliferative effect of celastrol on cultured cells --- p.142 / Chapter 4.2.4 --- Discussion --- p.145 / Chapter 4.3 --- Induction of Apoptosis by Celastrol on Human Keratinocytes --- p.146 / Chapter 4.3.1 --- Introduction --- p.146 / Chapter 4.3.2 --- Materials and methods --- p.146 / Chapter 4.3.2.1 --- Chemicals --- p.146 / Chapter 4.3.2.2 --- General cell culture --- p.147 / Chapter 4.3.2.3 --- Cell cycle analysis with PI staining --- p.147 / Chapter 4.3.2.4 --- Detection of apoptosis by flow cytometry --- p.147 / Chapter 4.3.2.5 --- Measurement of the mitochondrial membrane potential (ΔΨm) --- p.148 / Chapter 4.3.2.6 --- Western blot analysis --- p.148 / Chapter 4.3.2.7 --- Statistical analysis --- p.148 / Chapter 4.3.3 --- Results --- p.149 / Chapter 4.3.3.1 --- Induction of sub-G1 phase by celastrol on HaCaT cells --- p.149 / Chapter 4.3.3.2 --- Quantitative analysis of apoptotic cells by Annexin V-PI staining --- p.151 / Chapter 4.3.3.3 --- Alteration of ΔΨm --- p.153 / Chapter 4.3.3.4 --- Activation of caspase family protein --- p.155 / Chapter 4.3.3.5 --- Celastrol regulates the Bcl-2 family members --- p.159 / Chapter 4.3.4 --- Discussion --- p.161 / Chapter 4.4 --- Inhibition of NF-κB Transcription Factor Activation by Celastrol --- p.164 / Chapter 4.4.1 --- Introduction --- p.164 / Chapter 4.4.2 --- Materials and methods --- p.165 / Chapter 4.4.2.1 --- Chemicals --- p.165 / Chapter 4.4.2.2 --- General cell cultrue --- p.165 / Chapter 4.4.2.3 --- Western blot analysis --- p.165 / Chapter 4.4.2.4 --- Detect nuclear p65 by ELISA assay --- p.166 / Chapter 4.4.2.5 --- Statistical analysis --- p.166 / Chapter 4.4.3 --- Results --- p.167 / Chapter 4.4.3.1 --- Celastrol inhibited the NF-κB activation --- p.167 / Chapter 4.4.4 --- Discussion --- p.170 / Chapter 4.5 --- Induction of Terminal Differentiation by Celastrol --- p.173 / Chapter 4.5.1 --- Introduction --- p.173 / Chapter 4.5.2 --- Materials and methods --- p.174 / Chapter 4.5.2.1 --- Chemicals --- p.174 / Chapter 4.5.2.2 --- General cell culture --- p.174 / Chapter 4.5.2.3 --- CE formation assay --- p.174 / Chapter 4.5.2.4 --- Western blot analysis --- p.174 / Chapter 4.5.2.5 --- Statistical analysis --- p.174 / Chapter 4.5.3 --- Results --- p.175 / Chapter 4.5.3.1 --- Regulation of CE formation by celastrol --- p.175 / Chapter 4.5.3.2 --- Modulation of terminal differentiation markers by celastrol --- p.178 / Chapter 4.5.4 --- Discussion --- p.181 / Chapter 4.6 --- General Discussion --- p.183 / Chapter Chapter Five --- In vivo Anti-psoriatic Effects of Topical Preparation of 1-hydroxy-3-methyl-anthraquinone / Chapter 5.1 --- Introduction --- p.187 / Chapter 5.2 --- Material and Methods --- p.191 / Chapter 5.2.1 --- Chemicals --- p.191 / Chapter 5.2.2 --- Formulation of topical preparation containing HMA --- p.191 / Chapter 5.2.3 --- Mouse tail model --- p.192 / Chapter 5.2.4 --- Histopathological evaluation --- p.193 / Chapter 5.2.5 --- Statistical analysis --- p.194 / Chapter 5.3 --- Results --- p.195 / Chapter 5.3.1 --- Body weight profile --- p.195 / Chapter 5.3.2 --- Histological resutls --- p.197 / Chapter 5.4 --- Discussion --- p.201 / Chapter Chapter Six --- General Conclusions and Future Perspectives / Chapter 6.1 --- General Conclusions --- p.205 / Chapter 6.2 --- Future Perspectives --- p.210 / References / References by alphabetical order --- p.213

銀屑病是一種慢性炎症性皮膚病，其發病率約佔全球1-3%的人口。銀屑病的病理特徵包括角質細胞增殖和分化異常，同時伴隨炎症反應，白細胞聚集於真皮和表皮以及血管擴張。證據顯示角質細胞能參與及延續免疫反應，以達致維持或促進該病的作用。研究亦建議角質細胞減少凋亡是引致銀屑病的一個特定現象；因此，長期以來誘導角質細胞凋亡就被用作為治療銀屑病的一種有效策略。 / 根據銀屑病的嚴重程度，治療方法可分為三級：外用藥物主要用於比較輕微的病患，而光療適合中等程度的病患；對於嚴重病例則可使用系統性治療或生物製劑。基於大約75%的銀屑病患者屬於輕微至中度病患，外用藥物是目前應用最為廣泛的治療方法。在中國銀屑病治療的歷史中曾經使用過中草藥，研究亦表明，其治療機制可能通過抑制角質細胞增殖和誘導角質細胞凋亡。比較研究也指出，傳統中藥比西藥的副作用相對較少，及具有較長的舒緩期和較低的復發率。 / 我們先前的研究發現，茜草根提取物能夠抑制一個和銀屑病相關的HaCaT角質細胞增殖。本研究證實，茜草根的乙酸乙酯提取物（EA）能誘導HaCaT細胞凋亡，其抑制角質細胞增殖的作用比茜草根的乙醇提取物（EE）更為有效，並可和一個流行於歐洲國家的重要外用銀屑病治療藥地蒽酚相比。另外，透過不同的檢測，包括形態學觀察，細胞凋亡雙染（磷脂結合蛋白V-碘化丙啶）分析，細胞週期分析，去氧核醣核酸斷裂測試，原位末端轉移酶標記技術，免疫熒光染色以及西方墨點法，我們發現一種在茜草中的化合物，1,4-二羥基-2-萘甲酸（DHNA）能通過死亡受體介導，線粒體介導或不依賴胱天蛋白酶的途徑導致HaCaT細胞凋亡。同時，在其中一種銀屑病動物模型，小鼠鼠尾鱗片表皮上的初步研究顯示DHNA亦可誘導角質細胞分化。此外，在細胞水平（存活率，釋放白细胞介素-1α）和動物上（Draize動物皮膚刺激性試驗）的實驗結果表明DHNA比地蒽酚的刺激性較小。 / 總括而言，本研究透過人類皮膚細胞和動物實驗說明EA和DHNA的細胞凋亡機制，以及DHNA對皮膚的潛在刺激性。這些結果顯示EA和DHNA有潛能發展成為安全及能有效治療銀屑病的替代藥物。EA和DHNA可在一個連續療程中結合使用，其中EA藥效媲美地蒽酚，應能迅速清除銀屑病皮損；而DHNA比地蒽酚的刺激性小，則比較適合應用在這個連續療程中後來的維護保養階段 / Psoriasis is a chronic inflammatory skin disorder that affects approximately 1-3% of the population worldwide. It is characterized by epidermal hyperplasia or abnormal differentiation, infiltration of leucocytes into the dermis and epidermis, dilation of blood vessels in dermis and inflammation. Evidence indicates keratinocytes contributed to the disease, and keratinocytes also participate in maintaining the chronically perpetuating immune response that sustains psoriasis. Decrease in keratinocytes apoptosis is suggested to be a specific pathogenic phenomenon, and induction of keratinocytes apoptosis have long been considered as an effective anti-psoriatic strategy. / Treatment of psoriasis is based on disease severity. Topical agents are predominantly for mild conditions; phototherapy for moderate conditions and systemic treatment or biological agents for severe cases. Topical treatment remains the most widely used method as an estimated 75% of psoriatic patients have mild to moderate disease. Chinese herbs have been used for the treatment of psoriasis in China, and studies showed their mechanism on treating psoriasis may through inhibition of keratinocyte proliferation and induction of apoptosis. Comparison studies also show that traditional Chinese medicine has relatively fewer side effects than western therapeutic agents, with a longer remission time and lower recurrence rate. / The extract of the root of Rubia cordifolia L. (Rubiae Radix et Rhizoma) was previously found to inhibit keratinocyte proliferation using a psoriasis-relevant HaCaT cells model. In this study, the ethyl acetate extract of the root of Rubia cordifolia L. (EA) was confirmed to induce apoptosis on HaCaT cell, and the antiproliferative effect of EA is more potent than the ethanol extract of the herb (EE) and is comparable to dithranol, an important and popular topical treatment for psoriasis among Europe countries. Besides, we identified one of the components in Rubia cordifolia L., 1,4-dihydroxy-2-naphthoic acid (DHNA), could induce HaCaT keratinocyte apoptosis through the death receptor and mitochondria mediated pathway as well as in a caspase independent manner using various assays such as morphological examination, annexin V-PI staining, cell cycle analysis, DNA fragmentation, TUNEL assay, immunofluorescence staining and Western blot analysis. Moreover, DHNA was found to induce keratinocyte differentiation in a preliminary study using the in vivo mouse tail model of psoriasis. Furthermore, results from in vitro (cell viability, IL-1α release) and in vivo (Draize animal skin irritation test) experiments suggested DHNA have less irritation problems than dithranol. / In summary, this study describes the apoptotic mechanism of EA and DHNA, as well as the irritation potential of DHNA using different human skin cells and animal model. These results suggest EA and DHNA have the potential to develop as safe and effective therapeutic alternative for the treatment of psoriasis. EA and DHNA can be used together in a sequential therapy, in which EA is effective in rapid clearing of psoriatic lesions as its potency is comparable to dithranol; whereas DHNA is better suited for the later maintenance therapy for its milder irritation effect compared with dithranol. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Mok, Chong Fai. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 164-183). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract (English version) --- p.iv / Abstract (Chinese version) --- p.vi / List of Publication and Presentation --- p.viii / Acknowledgements --- p.ix / Table of Contents --- p.x / List of Tables --- p.xvi / List of Figures --- p.xvii / List of Abbreviations --- p.xx / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1. --- Psoriasis --- p.1 / Chapter 1.1.1. --- Histological features --- p.2 / Chapter 1.1.2. --- Role of keratinocytes in psoriasis --- p.4 / Chapter 1.1.3. --- Decrease in skin cell apoptosis --- p.8 / Chapter 1.2. --- Treatment of psoriasis --- p.9 / Chapter 1.2.1. --- Conventional treatment --- p.9 / Chapter 1.2.1.1. --- Mild disease --- p.10 / Chapter 1.2.1.1.1. --- Corticosteroids --- p.10 / Chapter 1.2.1.1.2. --- Vitamin D₃ analogs --- p.11 / Chapter 1.2.1.1.3. --- Tazarotene --- p.11 / Chapter 1.2.1.1.4. --- Anthralin --- p.12 / Chapter 1.2.1.1.5. --- Coal tar --- p.12 / Chapter 1.2.1.2. --- Moderate disease --- p.12 / Chapter 1.2.1.2.1. --- Phototherapy --- p.12 / Chapter 1.2.1.3. --- Severe disease --- p.13 / Chapter 1.2.1.3.1. --- Retinoids --- p.13 / Chapter 1.2.1.3.2. --- Methotrexate --- p.14 / Chapter 1.2.1.3.3. --- Cyclosporine --- p.14 / Chapter 1.2.1.3.4. --- Fumaric acid --- p.15 / Chapter 1.2.1.3.5. --- Biological agents --- p.15 / Chapter 1.2.2. --- Alternative treatment --- p.16 / Chapter 1.2.2.1. --- Traditional Chinese Medicine (TCM) --- p.17 / Chapter 1.3. --- Aims and objectives of the present study --- p.19 / Chapter Chapter 2: --- Apoptotic Action of Ethyl Acetate Fraction of the Root of Rubia cordifolia L. (Rubiae Radix et Rhizoma) on HaCaT Human Keratinocytes --- p.21 / Chapter 2.1. --- Introduction --- p.21 / Chapter 2.1.1. --- Rubia cordifolia L. --- p.21 / Chapter 2.1.2. --- Apoptosis --- p.22 / Chapter 2.1.3. --- Study objectives --- p.28 / Chapter 2.2. --- Materials and Methods --- p.30 / Chapter 2.2.1. --- Sources of medicinal materials --- p.30 / Chapter 2.2.2. --- Preparation of extracts --- p.30 / Chapter 2.2.3. --- Reagents --- p.31 / Chapter 2.2.4. --- Cell culture --- p.31 / Chapter 2.2.5. --- Proliferation assay --- p.32 / Chapter 2.2.6. --- Fluorescent staining for morphological evaluation --- p.33 / Chapter 2.2.7. --- Annexin V/propidium iodide staining --- p.33 / Chapter 2.2.8. --- JC-1 staining --- p.34 / Chapter 2.2.9. --- Statistical analysis --- p.35 / Chapter 2.3. --- Results --- p.36 / Chapter 2.3.1. --- EA inhibits proliferation of human epidermal HaCaT keratinocytes --- p.36 / Chapter 2.3.2. --- Alteration of cellular morphology --- p.39 / Chapter 2.3.3. --- EA increases phosphatidylserine externalization in HaCaT cells --- p.41 / Chapter 2.3.4. --- EA decreases MMP --- p.45 / Chapter 2.4. --- Discussion --- p.47 / Chapter Chapter 3: --- Identification of Pure Compound for Possible Apoptotic Action on HaCaT Human Keratinocytes and Detailed Mechanistic Study --- p.51 / Chapter 3.1. --- Introduction --- p.51 / Chapter 3.1.1. --- Anthraquinone --- p.51 / Chapter 3.1.2. --- Study objectives --- p.52 / Chapter 3.2. --- Materials and Methods --- p.54 / Chapter 3.2.1. --- Reagents --- p.54 / Chapter 3.2.2. --- Cell culture --- p.54 / Chapter 3.2.3. --- Proliferation assay --- p.55 / Chapter 3.2.4. --- Fluorescent staining for morphological evaluation --- p.56 / Chapter 3.2.5. --- Annexin V/propidium iodide staining --- p.56 / Chapter 3.2.6. --- JC-1 staining --- p.56 / Chapter 3.2.7. --- Cell cycle analysis --- p.56 / Chapter 3.2.8. --- Detection of DNA fragmentation --- p.57 / Chapter 3.2.9. --- Terminal Deoxynucleotidyltransferase-Mediated dUTP Nick End Labeling (TUNEL) assay --- p.57 / Chapter 3.2.10. --- Western blot analysis --- p.58 / Chapter 3.2.11. --- Immunofluorescence staining --- p.59 / Chapter 3.2.12. --- Statistical analysis --- p.60 / Chapter 3.3. --- Results --- p.61 / Chapter 3.3.1. --- DHNA inhibits proliferation of human epidermal HaCaT Keratinocytes --- p.61 / Chapter 3.3.2. --- Alteration of cellular morphology --- p.70 / Chapter 3.3.3. --- DHNA increases phosphatidylserine externalization in HaCaT cells --- p.72 / Chapter 3.3.4. --- DHNA decreases MMP --- p.76 / Chapter 3.3.5. --- DHNA causes G0/G1 cell cycle arrest in HaCaT cells --- p.78 / Chapter 3.3.6. --- DHNA increases DNA fragmentation --- p.81 / Chapter 3.3.7. --- DHNA increases TUNEL positive cells in HaCaT cells --- p.83 / Chapter 3.3.8. --- Western blot analysis --- p.85 / Chapter 3.3.9. --- DHNA induced Fas aggregation in HaCaT cells --- p.88 / Chapter 3.3.10. --- Caspase inhibition assay --- p.90 / Chapter 3.3.11. --- DHNA induced caspase independent apoptosis in HaCaT cells --- p.93 / Chapter 3.3.12. --- Effects of DHNA on MAPK in HaCaT cells --- p.96 / Chapter 3.3.13. --- MAPK inhibition assay --- p.100 / Chapter 3.4. --- Discussion --- p.104 / Chapter Chapter 4: --- Anti-Psoriatic Effects of Topical 1,4-Dihydroxy-2-naphthoic acid Formulation on in vivo Mouse Tail Experiments --- p.111 / Chapter 4.1. --- Introduction --- p.111 / Chapter 4.1.1. --- Keratinocytes differentiation process --- p.111 / Chapter 4.1.2. --- Animal model for psoriasis --- p.114 / Chapter 4.1.3. --- Study objectives --- p.119 / Chapter 4.2. --- Materials and Methods --- p.122 / Chapter 4.2.1. --- Reagents --- p.122 / Chapter 4.2.2. --- Formulation and preparation of topical drug --- p.122 / Chapter 4.2.3. --- Mice for in vivo experiments --- p.123 / Chapter 4.2.4. --- Treatment with topical preparations --- p.124 / Chapter 4.2.5. --- Statistical analysis --- p.125 / Chapter 4.3. --- Results --- p.126 / Chapter 4.3.1. --- Tail skin appearance after topical treatment --- p.126 / Chapter 4.3.2. --- Histological examination and findings --- p.128 / Chapter 4.4. --- Discussion --- p.132 / Chapter Chapter 5: --- Prediction of Skin Irritation Potential of 1,4-Dihydroxy-2-naphthoic acid by in vitro and in vivo Experiments --- p.135 / Chapter 5.1. --- Introduction --- p.135 / Chapter 5.1.1. --- Skin irritation --- p.135 / Chapter 5.1.2. --- Viability test and IL-1α release --- p.136 / Chapter 5.1.3. --- Animal irritation test --- p.139 / Chapter 5.1.4. --- Study objectives --- p.139 / Chapter 5.2. --- Materials and Methods --- p.141 / Chapter 5.2.1. --- Reagents --- p.141 / Chapter 5.2.2. --- Cell culture --- p.141 / Chapter 5.2.3. --- Viability test --- p.141 / Chapter 5.2.4. --- IL-1α release assay --- p.142 / Chapter 5.2.5. --- Animal irritation test --- p.142 / Chapter 5.2.6. --- Statistical analysis --- p.143 / Chapter 5.3. --- Results --- p.144 / Chapter 5.3.1. --- Viability test --- p.144 / Chapter 5.3.2. --- IL-1α release assay --- p.144 / Chapter 5.3.3. --- Animal irritation test --- p.147 / Chapter 5.4. --- Discussion --- p.152 / Chapter Chapter 6: --- General Discussion and Conclusions --- p.155 / References --- p.164

In den letzten Jahrzehnten haben sich die so genannten Biologika auch zur Therapie der Psoriasis etabliert. Zu diesen Medikamenten gehört auch Adalimumab, welches als vollständig humaner Antikörper eines der Schlüsselzytokine in der Pathogenese der Psoriasis, TNFα, neutralisiert. Allerdings führt die Therapie nicht bei allen Patienten zu ausreichendem Wirkerfolg. Da bisher vor Beginn der Therapie nicht zwischen den Patienten, die von der Therapie profitieren, und denen, die keine ausreichende Wirkung erfahren, unterschieden werden kann, werden die letzteren unnötigerweise den Risiken und Nebenwirkungen dieser Therapie ausgesetzt. In dieser Arbeit wurden die Interaktionen kultivierter Endothelzellen und Lymphozyten ex vivo unter dem Einfluss von Adalimumab untersucht. Insbesondere auf mögliche Unterschiede zwischen „Respondern“ und „Non-Respondern“ wurde im Hinblick auf die mögliche Entwicklung eines prädiktiven Tests für das Ansprechen auf Adalimumab ein Schwerpunkt gelegt. Lymphozyten gesunder Probanden und von Psoriasis-Patienten wurden ex vivo hinsichtlich ihrer Interaktion mit kultivierten Endothelzellen (HUVEC), mit und ohne TNFα-Stimulation, untersucht. Hierbei wurden sowohl frisch isolierte als auch kryokonservierte Lymphozyten verwendet, da sich zwischen diesen keine Unterschiede in den funktionellen Flusskammer-Versuchen zeigten. Nach Stimulation der Endothelzellen mit TNFα kam es zu einem deutlichen Anstieg des Rollens und der festen Adhäsion aller Lymphozyten an den Endothelzellen. Allerdings zeigten sich im Ausmaß dieser Interaktion deutliche inter-individuelle Unterschiede. Obwohl diese auch bei Psoriasis-Patienten auftraten, konnten keine signifikanten Unterschiede zwischen „Respondern“ und „Non-Respondern“ beobachtet werden. In den Untersuchungen zum Einfluss von Adalimumab auf TNFα-stimulierte Endothelzellen sowie die Interaktion dieser mit Lymphozyten ex vivo zeigte sich eine deutliche Abhängigkeit der Auswirkung vom Zeitpunkt der Behandlung. Erfolgte die Adalimumab-Behandlung vor oder gleichzeitig mit der TNFα-Stimulation der Endothelzellen, kam es zur Aufhebung der TNFα-induzierten Effekte sowohl in der Transkription der Adhäsionsmoleküle (PCR), der Expression dieser (Immunfluoreszenz-Mikroskopie) sowie der dynamischen Interaktionen mit Lymphozyten (Flusskammer). Eine dem TNFα nachfolgende Behandlung blieb ohne Wirkung, sodass davon auszugehen ist, dass sich die bereits induzierten Prozesse nicht mehr rückgängig machen lassen. Hier könnte eine Erklärung für das späte Eintreten der Adalimumab-Wirkung in vivo liegen. Somit konnten in dieser Arbeit vier zentrale Ergebnisse erzielt werden: Erstens, es wurde erstmals gezeigt, dass wichtige funktionelle Eigenschaften humaner Lymphozyten während der Kryokonservierung erhalten bleiben. Zweitens, es wurden erstmals deutliche inter-individuelle Unterschiede im Ausmaß der Interaktion zwischen Lymphozyten ex vivo mit TNFα-stimulierten Endothelzellen nachgewiesen. Adalimumab unterdrückte diese dynamischen Interaktionen, sofern seine Zugabe vor oder gleichzeitig mit der TNF-Exposition erfolgte. Drittens, diese inter-individuellen Unterschiede bestanden gleichermaßen bei gesunden Probanden und Psoriasis-Patienten. Viertens, die funktionellen Unterschiede erlaubten keine Unterscheidung zwischen Psoriasis-Patienten, deren Erkrankung sich durch Adalimumab besserte („Responder“), und denen, deren Erkrankung nicht auf diese Therapie ansprach („Non-Responder“).

Psoriasis is not an illness just of these days. 3-7% of world population suffers from this illness. This diploma work focuses on quality of life of ill people. We guess that it influences such a client in a holistic way. This illness interferes into personal, professional and social life. So psoriasis is not just an illness but also an everyday problem or handicap for those who get ill.Two questionnaires were made for this research, where first of those is focused on quality of life by a standard questionnaire and the second one was created on the base of analysis of D. Johnson model. Those questionnaires were given out to ill in South and Central Counties of Bohemia. The respondents were to chose, after their completing, which one is more sensitive to problems implying from this illness. The results are graphically shown and then paid attention in chapter Discussion.Finally I would like to say, those hypotheses were acknowledged. Psoriasis influences life in a holistic way, a nurse can have a significant influence on well-being of a client with this illness in area of her nursing interventions and D. Johnson model is an effective tool for providing those ill with psoriasis with care.

Le psoriasis est une dermatose inflammatoire chronique affectant 2 à 3 % de la population européenne. Les cytokines pro-inflammatoires ont un rôle crucial dans la pathogénèse du psoriasis. Parmi les voies de signalisation cytokinique, l'étude de la voie JAK/STAT a conduit au développement de traitements systémiques efficaces. Cependant, les essais cliniques évaluant les inhibiteurs JAK/STAT impliquent en grande majorité des administrations orales, la voie topique restant marginale. Le développement de formulations innovantes pour application topique contenant des inhibiteurs JAK/STAT semble être une stratégie prometteuse dans le cadre du psoriasis. Les nanoparticules de poly(acide lactique) (NP PLA) développées au laboratoire ont été étudiées pour la délivrance topique de principes actifs. Ce sont des vecteurs efficaces, qui s'accumulent dans les follicules pileux. De plus, l'encapsulation d'actifs dans des NP PLA permet une libération spécifique au niveau du site d'action, et donc une réduction de la toxicité. L'objectif de ce travail est d'élaborer des formulations semi-solides de NP PLA contenant un inhibiteur JAK/STAT pour le traitement topique du psoriasis, tout en caractérisant un modèle in vivo de psoriasis induit par applications d'Imiquimod. Cette caractérisation des lésions psoriasiformes permettra l'évaluation in vivo des nanoformulations topiques contenant des inhibiteurs JAK/STAT. Cinq formulations ont été élaborées puis caractérisées afin de répondre aux attentes galéniques pour une application topique. L'intégrité des NP PLA a été vérifiée, et la pénétration/perméation de molécules modèles à travers de la peau de souris inflammée a été évaluée / Psoriasis is a chronic inflammatory skin disease, affecting 2 to 3 % of European population. Inflammatory cytokines play a crucial role in the pathogenesis of psoriasis. Among cytokines signaling pathways, JAK/STAT pathway has been widely investigated, leading to the development of efficient systemic agents. However, current clinical trials evaluating JAK/Sat inhibitors mainly involve oral administrations, with few investigations on topical route. Developing innovative drug delivery systems for topical application of JAK/STAT inhibitors seems a promising strategy for psoriasis treatment. Poly(lactic acid) nanoparticles (PLA NPs) developed in the laboratory have been widely investigated for topical drug delivery and are efficient carriers for local dermatotherapy, especially through hair follicles. Moreover, drug encapsulation in PLA NPs for topical delivery allows a specific delivery to the site of action, and thus a decreased toxicity.The aim of this work was to elaborate semi-solid formulations of PLA NPs containing JAK/STAT inhibitors for topical treatment of psoriasis, while characterizing an in vivo model of Imiquimod-induced psoriasis in mice. This characterization of psoriasis-like skin lesions in Imiquimod treated mice provided key tools for in vivo evaluation of topical nanoformulations containing JAK/STAT inhibitors. Five formulations have been developed and then characterized in order to meet galenic criteria for topical drug administration. PLA NPs integrity was assessed, and penetration/permeation profiles of model dugs through inflamed mice skin were determined

Psoriasis vulgaris ist eine chronisch-entzündliche Hauterkrankung, welche mit verdickten, erythematös-schuppigen Plaques und systemischer Inflammation einhergeht. Bisher veröffentlichte epidemiologische Studien, sowie Studien an Tiermodellen und Zellkulturen lassen vermuten, dass psoriatische Inflammation den Knochenstoffwechsel verändert. Um dieser Fragestellung nachzugehen, wurden in der vorliegenden Studie zwei Serummarker des Knochenstoffwechsels, das n-terminale Propeptid des Typ 1-Prokollagens (P1NP) sowie das c-terminale Telopeptid des Typ 1-Kollagens (CTX-I) anhand von 42 Patienten mit Psoriasis vulgaris im Vergleich zu 40 gesunden Kontroll-Probanden mittels ELISA, einem enzymgekoppelten Immunadsorptionstest, analysiert. Unsere Daten zeigen eine Verminderung des Knochenaufbaumarkers P1NP bei Psoriasis-Patienten unabhängig von Geschlecht, Alter und BMI im Vergleich zu gesunden Kontroll-Personen, wohingegen CTX-I als Knochenresorptionsmarker unbeeinflusst bleibt. Dies lässt einen Psoriasis vulgaris-assoziierten Netto-Verlust an Knochensubstanz bei vermindertem Knochenaufbau und gleichbleibendem Knochenabbau vermuten. Unsere Studie zeigt, dass sowohl bei männlichen als auch weiblichen Patienten mit Psoriasis vulgaris die Konzentrationen von P1NP signifikant verringert sind. Hinsichtlich der Beeinflussung dieser Marker durch das Körpergewicht zeigte die vorliegende Arbeit, dass sowohl bei übergewichtigen als auch normalgewichtigen Patienten mit Psoriasis vulgaris die P1NP-Konzentrationen erniedrigt sind. Nach Unterteilung der Patienten-Kohorte nach Geschlecht und BMI konnten bei den Patienten mit Psoriasis signifikant bzw. tendenziell verringerte P1NP Serumlevel festgestellt werden, sodass von einem geschlechter- sowie BMI-unabhängigen Effekt der Psoriasis vulgaris ausgegangen werden kann. Hierbei muss jedoch beachtet werden, dass durch die Aufteilung in Unter-Kohorten die Fallzahlen pro Gruppe zum Teil sehr klein werden und damit die statistische Aussagekraft eingeschränkt ist. Interessanterweise korrelieren die P1NP-Level weder mit der Erkrankungsschwere anhand des PASI (Psoriasis Severity Index), BSA (Body surface area) und PGA (Physican´s Global Assessment), noch mit der Erkrankungsdauer oder der Gesamtleukozytenzahl im Blut als Entzündungsparameter. Dies könnte bedeuten, dass bereits eine geringe psoriatische Entzündung beziehungsweise eine kurze Erkrankungsdauer in den Knochenstoffwechsel eingreift und den Knochenaufbau hemmt. Da Psoriasis-Patienten Risikofaktoren für reduzierte Knochendichte aufweisen, wie beispielsweise hohe Raten an Tabakkonsum sowie geringer sportlicher Aktivität, erfolgte eine Analyse der Knochenresorptionsmarker in entsprechenden Unter-Kohorten. Dennoch zeigten sich keine signifikanten Unterschiede bezüglich der P1NP- und CTX-I-Serumwerte zwischen Psoriasis vulgaris Patienten mit/ohne Tabakkonsum und aktivem/inaktivem Lebensstil. In Zusammenschau unserer hier vorgestellten Daten und anderen bisher veröffentlichten epidemiologischen Studien sowie Studien an Tiermodellen und Zellkulturen welche unsere Ergebnisse stützen, lässt sich vermuten, dass die psoriatische Entzündung den Knochenstoffwechsel verändert und damit möglicherweise das Osteoporose-Risiko erhöht. Folglich könnte eine konsequente anti-psoriatische Therapie sowie ein frühzeitiges Screening der Knochenqualität hilfreich sein, um psoriasis-assoziierten Knochenveränderungen vorzubeugen.:1 Einleitung ................................................................................................................ 1 1.1 Psoriasis .......................................................................................................... 1 1.1.1 Definition und klinisches Bild ................................................................. 1 1.1.2 Epidemiologie ........................................................................................ 1 1.1.3 Klassifikation .......................................................................................... 2 1.1.4 Histologie der Psoriasis vulgaris ............................................................ 7 1.1.5 Pathomechanismen ............................................................................... 9 1.1.6 Therapieformen ................................................................................... 12 1.1.7 Psoriasis-assoziierte Komorbiditäten .................................................. 18 1.2 Knochenstoffwechsel ................................................................................... 19 1.2.1 Definition und Physiologie ................................................................. 19 1.2.2 Knochenstoffwechselregulation ......................................................... 22 1.2.3 Zusammenhang zwischen Entzündung und Knochenstoffwechsel ... 27 2 Fragestellung ..................................................................................................... 30 3 Materialien und Methoden .................................................................................. 31 3.1 Klinische Methoden ..................................................................................... 31 3.1.1 Patientenselektion und Studiendesign ............................................... 31 3.1.2 Einschlusskriterien ............................................................................. 31 3.1.3 Ausschlusskriterien ............................................................................ 32 3.1.4 Kohortencharakterisierung ................................................................. 33 3.1.5 Studienablauf ..................................................................................... 35 3.1.6 Fragebogen ........................................................................................ 36 3.2 Labormethoden ............................................................................................ 37 3.3 Statistische Analyse ..................................................................................... 37 4 Ergebnisse .......................................................................................................... 38 4.1 P1NP-Serumkonzentrationen ...................................................................... 38 4.1.1 P1NP in Zusammenhang mit dem Geschlecht ................................. 39 4.1.2 P1NP in Zusammenhang mit dem BMI .............................................. 40 4.1.3 P1NP in Zusammenhang mit dem Geschlecht und BMI..................... 41 4.1.4 P1NP in Zusammenhang mit systemischer Therapie......................... 43 4.1.5 P1NP in Zusammenhang mit Krankheitsschwere, Erkrankungsdauer und Entzündungsparametern.....................................................................45 4.1.6 P1NP in Zusammenhang mit den äußeren Einflussfaktoren Tabakkonsum und inaktiver Lebensstil ...................................................................... 46 4.2 CTX-I-Serumkonzentrationen ................................................................ 47 4.2.1 CTX-I in Zusammenhang mit dem Geschlecht .................................. 48 4.2.3 CTX-I-Serumkonzentrationen in Zusammenhang mit dem Geschlecht und BMI ............................................................................................. 50 4.2.4 CTX-I in Zusammenhang mit systemischer Therapie ....................... 52 4.2.5 CTX-I in Zusammenhang mit Krankheitsschwere Erkrankungsdauer und Entzündungsparametern ............................................................54 4.2.6 CTX-I in Zusammenhang mit den äußeren Einflussfaktoren Tabakkonsum und inaktiver Lebensstil ............................................. 55 5 Diskussion .......................................................................................................... 56 6 Zusammenfassung ............................................................................................. 71 7 Literaturverzeichnis ............................................................................................ 73 8 Abbildungsverzeichnis ........................................................................................ 81 9 Tabellenverzeichnis ............................................................................................ 82 10 Anlagen ............................................................................................................. 83 10.1 Fragebogen der Studie .............................................................................. 83 10.2 Selbstständigkeitserklärung ....................................................................... 85 10.3 Lebenslauf ................................................................................................. 86 10.4 Danksagung .............................................................................................. 87

Bakgrund: Känsla av sammanhang (SOC) är en salutogen modell som beskriver hur människor ser livet och hanterar situationer med hjälp av resurser för att främja hälsa. Psoriasis är en kronisk och autoimmun sjukdom med en prevalens på två procent i Skandinavien. Den vanligaste formen av psoriasis är plackpsoriasis som karaktäriseras av rodnad, fjällning och förtjockning av epidermis. Syfte: Syftet var att belysa psykosociala aspekter av att leva med psoriasis. Metod: En allmän litteraturstudie där 11 resultatartiklar granskades i sin helhet och analyserades i enlighet med en databearbetningsmodell för litteraturöversikter med tre steg. Resultat: Genom dataanalysen identifierades tre huvudteman: Att skämmas över sin kropp, Att uppleva hinder i livet och Att hantera psoriasis. Resultatet påvisade flertalet psykosociala begränsningar som präglade vardagen och livskvaliteten negativt. Hälsofrämjande hanteringsstrategier identifierades dessutom. Konklusion: Ytterligare omvårdnadsforskning om levda erfarenheter hos personer med psoriasis behövs då området är begränsat utforskat. I vårdsammanhang bör personen ses som en helhet och inte enbart utifrån ett patogent perspektiv. / Background: Sence of coherence (SOC) is a salutogenic model that describe how people perceive life and deal with situations using their own resources to develop health. Psoriasis is a chronic and autoimmune disease with a prevalence of two percent in Scandinavia. The most common type of psoriasis is plaque psoriasis, which is characterized by raised skin patches, together with redness and scaling.  Aim: The aim was to illustrate the psychosocial aspects of living with psoriasis. Method: A general literature review in which 11 outcome articles were reviewed in their entirety and analyzed according to a three-step data processing model for literature reviews. Results: Through the data analysis, three main themes were identified: to be ashamed of one’s body, experiencing obstacles in life and dealing with psoriasis. The results revealed a large number of psychosocial limitations that negatively affected everyday life and quality of life. Health promoting coping strategies were also identified. Conclusion: Further nursing research is required to explore the lived life of people with psoriasis, as the current knowledge is limited. In the context of care, the person should be seen as a whole and not solely from a pathogenic perspective.