methylcellulose and Psoriasis

Human prolactin (PRL) is a well-known hormone for pituitary of lactation and reproduction, but it also has immunostimulatory effect in some inflammatory or autoimmune diseases including psoriasis, which has not been well elucidated. This study aimed to determine the relationship between PRL and psoriasis through clinical case-control studies, and explore the function of PRL in the pathogenesis of imiquimod (IMQ)-induced psoriasis-like mouse model. Serum from patients with psoriasis vulgaris (PsV), patients with erythrodermic psoriasis, and healthy controls (HCs) were collected for PRL test. Skin biopsies were collected for PRL, PRL receptors (PRLRs), cytokines mRNA level determination, PRL immunohistochemistry and PRL Western blotting. Mice were divided into four groups (each n = 6): control group (CON), IMQ group, anti-PRL group and solvent group. Anti-PRL group and solvent group mice were treated with PRL antagonist (cabergoline) and the solvent (0.25% methylcellulose) separately. The serum PRL level of PsV patients was significantly higher than that of HCs (P < 0.001). Compared with HCs, the mRNA levels of PRL and Th1/Th17 cytokines in skin lesions increased significantly (P < 0.05), and the PRL protein level was also significantly elevated in the epidermis and dermis of PsV patients. In IMQ-induced psoriasis-like mouse model, the mRNA and protein levels of PRL in skin lesions were significantly higher than CON group (P < 0.01). Comparing to solvent group, serum PRL level and PRL, cytokines mRNA levels in skin lesions all decreased significantly and the skin inflammatory condition was also alleviated obviously in anti-PRL group. This study suggests that local production of PRL is the main resource of PRL in skin lesions and may play an important role in skin inflammatory of psoriasis.

Topics: Adult; Aged; Animals; Cabergoline; Cytokines; Dermis; Disease Models, Animal; Dopamine Agonists; Epidermis; Female; Humans; Imiquimod; Male; Methylcellulose; Mice; Middle Aged; Prolactin; Psoriasis; Receptors, Prolactin; Risk Adjustment; RNA, Messenger; Solvents; TRPV Cation Channels

The aim of this study was to develop an effective drug delivery system for the simultaneous topical delivery of two anti-inflammatory drugs, spantide II (SP) and ketoprofen (KP). To achieve this primary goal, we have developed a skin permeating nanogel system (SPN) containing surface modified polymeric bilayered nanoparticles along with a gelling agent. Poly-(lactide-co-glycolic acid) and chitosan were used to prepare bilayered nanoparticles (NPS) and the surface was modified with oleic acid (NPSO). Hydroxypropyl methyl cellulose (HPMC) and Carbopol with the desired viscosity were utilized to prepare the nanogels. The nanogel system was further investigated for in vitro skin permeation, drug release and stability studies. Allergic contact dermatitis (ACD) and psoriatic plaque like model were used to assess the effectiveness of SPN. Dispersion of NPSO in HPMC (SPN) produced a stable and uniform dispersion. In vitro permeation studies revealed increase in deposition of SP for the SP-SPN or SP+KP-SPN in the epidermis and dermis by 8.5 and 9.5 folds, respectively than SP-gel. Further, the deposition of KP for KP-SPN or SP+KP-SPN in epidermis and dermis was 9.75 and 11.55 folds higher, respectively than KP-gel. Similarly the amount of KP permeated for KP-SPN or SP+KP-SPN was increased by 9.92 folds than KP-gel. The ear thickness in ACD model and the expression of IL-17 and IL-23; PASI score and TEWL values in psoriatic plaque like model were significantly less (p < 0.001) for SPN compared to control gel. Our results suggest that SP+KP-SPN have significant potential for the percutaneous delivery of SP and KP to the deeper skin layers for treatment of various skin inflammatory disorders.

Topics: Administration, Cutaneous; Aminoquinolines; Animals; Anti-Inflammatory Agents; Disease Models, Animal; Drug Delivery Systems; Humans; Hypromellose Derivatives; Imiquimod; Immunohistochemistry; Inflammation; Methylcellulose; Mice; Mice, Inbred C57BL; Nanogels; Nanoparticles; Particle Size; Permeability; Polyethylene Glycols; Polyethyleneimine; Psoriasis; Rheology; Skin; Surface Properties; Viscosity

During malignant transformation in skin, epidermal keratinocytes (KCs) frequently acquire the capacity to by-pass cellular senescence, a response that normally limits their unrestricted proliferation. Despite growing interest in the role for senescence during aging of skin and cutaneous carcinogenesis, little is known regarding regulation of three proteins encoded by the INK4a/ARF locus (p12, p14(ARF), p16) in KCs. In this study, several molecular pathways are explored using cultured KCs and KCs freshly isolated from psoriatic plaques. p16 and p14(ARF) are predominantly expressed spontaneously when foreskin-derived early-passage KCs undergo confluency-induced premature senescence. Induction of p14(ARF) on confluency occurred with low E2F-1 levels. Suspension of KCs in methylcellulose induced p12 expression. Addition of various cytokines (interferon-gamma, tumor necrosis factor-alpha) or a phorbol ester [12-O-tetradecanoylphorbol-13-acetate (TPA)] only induced p16, but not p14(ARF). Confluent KCs up-regulated Ras activity and the downstream signaling involving ERK. Addition of MAPK inhibitor blocked cytokine and TPA-induced p16 expression. Confluency and interferon-gamma induced premature senescence and p16 expression was linked to induction of the transcription factor Egr-1. KCs derived from chronic psoriatic plaques were characterized by enhanced p16, p14(ARF), and p12 expression accompanied by elevated Egr-1 levels. These results demonstrate that multiple and highly divergent stimuli can trigger the senescent checkpoint in human KCs with differential regulation of p16, p14(ARF), and p12. Although abnormal mitogenic signaling by oncogenic Ras is generally cited as being responsible for induction of premature senescence, our findings indicate that a broader perspective is warranted, to include confluency and cytokine-/TPA-induced pathways for KCs.

Topics: Cell Division; Cells, Cultured; Cellular Senescence; Cyclin-Dependent Kinase Inhibitor p16; Cytokines; DNA-Binding Proteins; Early Growth Response Protein 1; Enzyme Inhibitors; Gene Expression Regulation; Genes, cdc; Genes, Reporter; Humans; Immediate-Early Proteins; Keratinocytes; Methylcellulose; Mitogen-Activated Protein Kinases; Psoriasis; ras Proteins; Reference Values; Signal Transduction; Tetradecanoylphorbol Acetate; Transcription Factors; Transfection; Tumor Suppressor Protein p14ARF

We have previously shown that antisense oligonucleotides effectively reduced insulin-like growth factor I receptor expression in human psoriatic skin grafted on to nude mice when injected intradermally. We therefore investigated the penetration of C-5 propyne modified antisense oligonucleotides into human normal and psoriatic skin after topical administration. Oligonucleotide (37.5 microg; 250 microM) was applied in aqueous solution or 5% methylcellulose gel for 24 h, prior to live confocal microscopy and fluorescence microscopy of fixed sections. We found that oligonucleotide could penetrate through the stratum corneum of psoriatic but not normal human skin over large regions of the epidermis. The oligonucleotide was localized to the nucleus of large parakeratotic cells in the psoriatic skin as well as smaller basal and suprabasal keratinocytes. In normal human skin, oligonucleotide was confined to the stratum corneum, with little or no oligonucleotide apparent in the viable epidermis. Electrophoresis of oligonucleotide recovered from treated psoriatic and normal skin revealed that the oligonucleotide remained intact over the 24 h period. In summary, we found that C-5 propyne modified antisense oligonucleotides could reach the target cells (in this case basal keratinocytes) after topical administration to psoriatic but not normal skin.

Topics: Administration, Topical; Alkynes; Epidermis; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Gels; Genetic Therapy; Humans; In Vitro Techniques; Keratinocytes; Methylcellulose; Microscopy, Fluorescence; Oligonucleotides, Antisense; Psoriasis

Keratinocytes (KC) are important source of and targets for several cytokines. Although KC express IL-15 mRNA, the functional effects of IL-15 on these epithelial cells remain to be dissected. Investigating primary human foreskin KC and HaCaT cells, we show here by semiquantitative RT-PCR and flow cytometric analysis that both translate IL-15 and IL-15R mRNA and express IL-15 and IL-15Ralpha protein on the cell surface, suggesting that human KC can employ IL-15 for juxtacrine signaling. While IL-15 exerted no significant effect on KC proliferation and IL-6 or IL-8 secretion, IL-15 inhibited both anti-Fas and methylcellulose-induced KC apoptosis in vitro. This is in line with the recognized potent anti-apoptotic effects of IL-15. IL-2, whose receptor shares two components with the IL-15R, failed to inhibit KC apoptosis. Together with the role of IL-15 in sustaining chronic immune reactions, this invited the question of whether a reduction of KC apoptosis by IL-15 may be involved in the pathogenesis of psoriasis, a chronic hyperproliferative inflammatory skin disease characterized by abnormally low KC apoptosis in the epidermis. Remarkably, compared with nonlesional psoriatic skin and skin of healthy volunteers, lesional psoriatic epidermis showed high IL-15 protein expression in the epidermis and enhanced binding activity for IL-15. Therefore, antagonizing the inhibitory effects of IL-15 on KC apoptosis deserves exploration as a novel therapeutic strategy in psoriasis management.

Topics: Apoptosis; Binding Sites; Cell Division; Cell Membrane; Cells, Cultured; Epidermis; fas Receptor; Gene Expression Regulation; Humans; Immune Sera; Immunosuppressive Agents; Interleukin-15; Interleukin-6; Interleukin-8; Keratinocytes; Methylcellulose; Psoriasis; Receptors, Interleukin-15; Receptors, Interleukin-2; RNA, Messenger

Previously we observed that hyperplastic epidermal keratinocytes characteristic of psoriasis had abundant amounts of the cell survival protein Bcl-xL; however, whether this overexpression correlated with enhanced survival was unclear because the majority of epidermal cells possess nuclei that are positively labeled by an assay typically regarded as indicative of cells undergoing apoptosis (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) staining). To clarify this apparent discrepancy, we explored the propensity of keratinocytes derived from psoriatic plaques to undergo apoptosis and also determined the reliability of TUNEL staining as an indicator of apoptosis in keratinocytes in vitro and in vivo. First, a keratinocyte cell line, HaCat, was examined before and after being suspended in semisolid medium (methylcellulose) using flow cytometry to detect TUNEL-positive cells, and the percentage of positive cells was correlated to the presence or absence of double-stranded DNA fragmentation using pulsed field gel electrophoresis. After 18 hours in methylcellulose suspension, apoptosis was detected in HaCat cells when at least 5% of the cell population was undergoing programmed cell death. Second, we examined 23 clinical specimens of skin (13 from psoriatic patients and 10 from healthy control subjects) and observed that no double-stranded DNA fragmentation was present in any of the freshly isolated keratinocytes from either normal or psoriatic patients. Keratinocytes from 9 of 12 normal skin samples underwent double-stranded DNA fragmentation after being in methylcellulose for 18 to 24 hours, which contrasts with keratinocytes from lesions of psoriasis where only 1 of 13 of the skin samples had these changes. Third, two-color immunofluorescence staining of psoriatic plaques revealed that numerous TUNEL-positive keratinocytes were also positive for proliferating cell nuclear antigen and Ki-67 antigens and that by flow cytometry TUNEL-positive keratinocytes obtained from psoriatic plaques possessed a DNA content profile indicative of proliferating and not dying cells. These results demonstrate that keratinocytes within psoriatic plaques do not have double-stranded DNA breaks, that they have a prolonged capacity to resist induction of apoptosis compared with normal-skin-derived keratinocytes or cultured HaCat cells, and that caution is necessary for proper interpretation related to detection of 3'-OH DNA ends (ie, TUNEL positi

Topics: Apoptosis; Cell Line; DNA; DNA Fragmentation; Genetic Techniques; Humans; Keratinocytes; Ki-67 Antigen; Methylcellulose; Proliferating Cell Nuclear Antigen; Psoriasis; Reference Values; Skin

During terminal differentiation, basal keratinocytes lose gradually contact with the basement membrane, a process accompanied by the progressive functional down-regulation and loss of integrin expression. Understanding the molecular nature of this complex mechanism will eventually lead to insight into the pathogenesis of differentiation disorders of the epidermis, e.g. psoriasis.. The monoclonal antibody 8D9 against the very late antigen 5 (VLA-5) integrin subunit was used to study the expression and down-regulation of this protein in several experimental paradigms of keratinocyte differentiation.. Primary cultures of human keratinocytes were prepared and used as such, or after induction of terminal differentiation with methylcellulose and/or calcium. Expression of the 8D9 epitope was analyzed using immunoblotting, protein chemistry and immunocytochemistry on cultured cells and on skin biopsies from control and psoriatic patients.. The monoclonal antibody 8D9 reacts with the alpha 5-subunit of human VLA-5 integrin and with a 68-kD antigen that is strongly expressed in differentiating keratinocytes in vitro and in the cornified layers of human skin in vivo. Psoriatic skin showed additional immunoreactivity in the upper spinous and granular layers. Based on indirect immunological and chemical evidence we suggest that the 68-kD protein is an amino-terminal degradation product of the alpha 5-subunit, which provides a new and interesting marker of differentiating keratinocytes.

Topics: Antibodies, Monoclonal; Antigens, CD; Basement Membrane; Biopsy; Calcium; Cell Differentiation; Cells, Cultured; Down-Regulation; Epitopes; Gene Expression Regulation; Humans; Immunoblotting; Immunohistochemistry; Integrin alpha5; Integrin beta1; Keratinocytes; Methylcellulose; Molecular Weight; Psoriasis; Receptors, Fibronectin

Topics: Chloramphenicol; Epithelium; Lactose; Methacrylates; Methylcellulose; Pregnadienetriols; Psoriasis; Radiation Effects; Talc; Ultraviolet Rays; Wound Healing; Zinc Oxide