Histamine enhances the production of human β-defensin-2 in human keratinocytes

The anti-microbial peptide human β-defensin-2 (hBD-2), produced by epidermal keratinocytes, plays pivotal roles in anti-microbial defense, inflammatory dermatoses, and wound repair. hBD-2 induces histamine release from mast cells. We examined the in vitro effects of histamine on hBD-2 production in normal human keratinocytes. Histamine enhanced TNF-α- or IFN-γ-induced hBD-2 secretion and mRNA expression. Histamine alone enhanced transcriptional activities of NF-κB and activator protein-1 (AP-1) and potentiated TNF-α-induced NF-κB and AP-1 activities or IFN-γ-induced NF-κB and STAT1 activities. Antisense oligonucleotides against NF-κB components p50 and p65, AP-1 components c-Jun and c-Fos, or H1 antagonist pyrilamine suppressed hBD-2 production induced by histamine plus TNF-α or IFN-γ. Antisense oligonucleotide against STAT1 only suppressed hBD-2 production induced by histamine plus IFN-γ. Histamine induced serine phosphorylation of inhibitory NF-κBα (IκBα) alone or together with TNF-α or IFN-γ. Histamine induced c-Fos mRNA expression alone or together with TNF-α, whereas it did not further increase c-Jun mRNA levels enhanced by TNF-α. Histamine induced serine phosphorylation of STAT1 alone or together with IFN-γ, whereas it did not further enhance IFN-γ-induced tyrosine phosphorylation of STAT1. The histamine-induced serine phosphorylation of STAT1 was suppressed by MAPKK (MEK) inhibitor PD98059. These results suggest that histamine stimulates H1 receptor and potentiates TNF-α- or IFN-γ-induced hBD-2 production dependent on NF-κB, AP-1, or STAT1 in human keratinocytes. Histamine may potentiate anti-microbial defense, skin inflammation, and wound repair via the induction of hBD-2.

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Leptin Enhances Human β-Defensin-2 Production in Human Keratinocytes

Endocrinology ◽

10.1210/en.2008-0343 ◽

2008 ◽

Vol 149 (10) ◽

pp. 5189-5198 ◽

Cited By ~ 33

Author(s):

Naoko Kanda ◽

Shinichi Watanabe

Keyword(s):

Mrna Expression ◽

Tyrosine Phosphorylation ◽

P38 Mapk ◽

Wound Repair ◽

Human Keratinocytes ◽

Janus Kinase ◽

Serine Phosphorylation ◽

Epidermal Keratinocytes ◽

Antimicrobial Defense

Leptin, an adipocyte-derived cytokine/hormone, modulates innate and adaptive immunity. Human β-defensin-2 (hBD-2) produced by epidermal keratinocytes promotes cutaneous antimicrobial defense, inflammation, and wound repair. We examined the in vitro effects of leptin on hBD-2 production in human keratinocytes. hBD-2 secretion and mRNA expression were analyzed by ELISA and RT-PCR, respectively. Although leptin alone was ineffective, it enhanced IL-1β-induced hBD-2 secretion and mRNA expression in keratinocytes. IL-1β- and IL-1β plus leptin-induced hBD-2 production both were suppressed by antisense oligonucleotides against nuclear factor-κB (NF-κB) p50 and p65; the latter was also suppressed by antisense signal transducer and activator of transcription (STAT)1 and STAT3. IL-1β enhanced the transcriptional activity of NF-κB, whereas leptin enhanced STAT1 and STAT3 activity. The p38 MAPK inhibitor SB202190 suppressed IL-1β- and IL-1β plus leptin-induced hBD-2 production, IL-1β-induced NF-κB activity, and leptin-induced STAT1 and STAT3 activity; contrastingly, the Janus kinase (JAK) 2 inhibitor AG490 suppressed IL-1β plus leptin-induced hBD-2 production and leptin-induced STAT1 and STAT3 activity. IL-1β induced serine phosphorylation of inhibitory κBα, STAT1, and STAT3. Leptin induced tyrosine and serine phosphorylation of STAT1 and STAT3, both of which were suppressed by AG490, and serine phosphorylation was also suppressed by SB202190. IL-1β or leptin individually induced threonine/tyrosine phosphorylation of p38 MAPK, whereas only leptin induced tyrosine phosphorylation of JAK2, suggesting that leptin may enhance hBD-2 production in keratinocytes by activating STAT1 and STAT3 via JAK2 and p38 MAPK in cooperation with NF-κB, which is activated by IL-1β. Leptin may promote cutaneous antimicrobial defense, inflammation, and wound repair via hBD-2.

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Water Extract of Rubus coreanus Prevents Inflammatory Skin Diseases In Vitro Models

Plants ◽

10.3390/plants10061230 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1230

Author(s):

Sumin Pyeon ◽

Ok-Kyung Kim ◽

Ho-Geun Yoon ◽

Shintae Kim ◽

Kyung-Chul Choi ◽

...

Keyword(s):

Mrna Expression ◽

Skin Diseases ◽

Water Extract ◽

Total Phenolic ◽

Hacat Cells ◽

Rubus Coreanus ◽

Tnf Α ◽

Ifn Γ ◽

Inflammatory Skin

Atopic dermatitis (AD) is a chronic inflammatory skin disease caused by immune hypersensitivity reaction. The cause of AD is unclear, but its symptoms have a negative effect on quality of life; various treatment methods to alleviate these symptoms are underway. In the present study, we aimed to evaluate in vitro antioxidant and anti-inflammatory effects of Rubus coreanus water extract (RCW) on AD. Total phenolic compounds and flavonoid content of RCW were 4242.40 ± 54.84 mg GAE/g RCE and 1010.99 ± 14.75 mg CE/g RCW, respectively. RCW reduced intracellular reactive oxygen species level and increased the action of antioxidant enzymes, such as catalase, superoxide dismutase, and glutathione peroxidase in tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ)-stimulated HaCaT cells. Moreover, mRNA expression of the pro-inflammatory cytokines, including TNF-α, interleukin-1β, and interleukin-6, was downregulated by RCW in the TNF-α/IFN-γ-stimulated cells. The levels of inflammatory chemokines (thymus- and activation-regulated chemokine; eotaxin; macrophage-derived chemokine; regulated on activation, normal T-cell expressed and secreted; and granulocyte-macrophage colony-stimulating factor) and intercellular adhesion molecule-1 were decreased in the TNF-α/IFN-γ-stimulated HaCaT cells after RCW treatment. Additionally, the mRNA expression levels of filaggrin and involucrin, proteins that form the skin, were increased by RCW. Furthermore, RCW inhibited the nuclear factor kappa-light-chain-enhancer of the activated B cells pathway in the TNF-α/IFN-γ-stimulated HaCaT cells. Collectively, the present investigation indicates that RCW is a potent substance that inhibits AD.

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High Matrix Metalloproteinase Production Correlates with Immune Activation and Leukocyte Migration in Leprosy Reactional Lesions

Infection and Immunity ◽

10.1128/iai.00896-09 ◽

2009 ◽

Vol 78 (3) ◽

pp. 1012-1021 ◽

Cited By ~ 17

Author(s):

Rosane M. B. Teles ◽

Rose B. Teles ◽

Thais P. Amadeu ◽

Danielle F. Moura ◽

Leila Mendonça-Lima ◽

...

Keyword(s):

Matrix Metalloproteinase ◽

Cultured Cells ◽

Leukocyte Migration ◽

Therapeutic Interventions ◽

Matrix Metalloproteinase 2 ◽

Mrna Levels ◽

Membrane Breakdown ◽

Tnf Α ◽

Ifn Γ

ABSTRACT Gelatinases A and B (matrix metalloproteinase 2 [MMP-2] and MMP-9, respectively) can induce basal membrane breakdown and leukocyte migration, but their role in leprosy skin inflammation remains unclear. In this study, we analyzed clinical specimens from leprosy patients taken from stable, untreated skin lesions and during reactional episodes (reversal reaction [RR] and erythema nodosum leprosum [ENL]). The participation of MMPs in disease was suggested by (i) increased MMP mRNA expression levels in skin biopsy specimens correlating with the expression of gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α), (ii) the detection of the MMP protein and enzymatic activity within the inflammatory infiltrate, (iii) increased MMP levels in patient sera, and (iv) the in vitro induction of MMP-9 by Mycobacterium leprae and/or TNF-α. It was observed that IFN-γ, TNF-α, MMP-2, and MMP-9 mRNA levels were higher in tuberculoid than lepromatous lesions. In contrast, interleukin-10 and tissue inhibitor of MMP (TIMP-1) message were not differentially modulated. These data correlated with the detection of the MMP protein evidenced by immunohistochemistry and confocal microscopy. When RR and ENL lesions were analyzed, an increase in TNF-α, MMP-2, and MMP-9, but not TIMP-1, mRNA levels was observed together with stronger MMP activity (zymography/in situ zymography). Moreover, following in vitro stimulation of peripheral blood cells, M. leprae induced the expression of MMP-9 (mRNA and protein) in cultured cells. Overall, the present data demonstrate an enhanced MMP/TIMP-1 ratio in the inflammatory states of leprosy and point to potential mechanisms for tissue damage. These results pave the way toward the application of new therapeutic interventions for leprosy reactions.

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Aberrant MUC1 accumulation in salivary glands of Sjögren’s syndrome patients is reversed by TUDCA in vitro

Rheumatology ◽

10.1093/rheumatology/kez316 ◽

2019 ◽

Vol 59 (4) ◽

pp. 742-753 ◽

Cited By ~ 3

Author(s):

Isabel Castro ◽

Nicolás Albornoz ◽

Sergio Aguilera ◽

María-José Barrera ◽

Sergio González ◽

...

Keyword(s):

Er Stress ◽

Salivary Glands ◽

Nuclear Translocation ◽

Secretory Process ◽

Mrna Levels ◽

Mucin 1 ◽

Tnf Α ◽

Mucin Expression ◽

Ifn Γ

Abstract Objectives Xerostomia in SS patients has been associated with low quality and quantity of salivary mucins, which are fundamental for the hydration and protection of the oral mucosa. The aim of this study was to evaluate if cytokines induce aberrant mucin expression and whether tauroursodeoxycholic acid (TUDCA) is able to counteract such an anomaly. Methods Labial salivary glands from 16 SS patients and 15 control subjects, as well as 3D acini or human submandibular gland cells stimulated with TNF-α or IFN-γ and co-incubated with TUDCA, were analysed. mRNA and protein levels of Mucin 1 (MUC1) and MUC7 were determined by RT-qPCR and western blot, respectively. Co-immunoprecipitation and immunofluorescence assays for mucins and GRP78 [an endoplasmic reticulum (ER)-resident protein] were also performed. mRNA levels of RelA/p65 (nuclear factor-κB subunit), TNF-α, IL-1β, IL-6, SEL1L and EDEM1 were determined by RT-qPCR, and RelA/p65 localization was evaluated by immunofluorescence. Results MUC1 is overexpressed and accumulated in the ER of labial salivary gland from SS patients, while MUC7 accumulates throughout the cytoplasm of acinar cells; however, MUC1, but not MUC7, co-precipitated with GRP78. TUDCA diminished the overexpression and aberrant accumulation of MUC1 induced by TNF-α and IFN-γ, as well as the nuclear translocation of RelA/p65, together with the expression of inflammatory and ER stress markers in 3D acini. Conclusion Chronic inflammation alters the secretory process of MUC1, inducing ER stress and affecting the quality of saliva in SS patients. TUDCA showed anti-inflammatory properties decreasing aberrant MUC1 accumulation. Further studies are necessary to evaluate the potential therapeutic effect of TUDCA in restoring glandular homeostasis in SS patients.

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IL-2, IL-5, TNF-α and IFN-γ mRNA expression in epidermal keratinocytes of systemic lupus erythematosus skin lesions

Clinics ◽

10.1590/s1807-59322011000100014 ◽

2011 ◽

Vol 66 (1) ◽

pp. 77-82 ◽

Cited By ~ 16

Author(s):

José Ronaldo M Carneiro ◽

Hellen T Fuzii ◽

Cristiane Kayser ◽

Fernando L Alberto ◽

Fernando A Soares ◽

...

Keyword(s):

Systemic Lupus Erythematosus ◽

Mrna Expression ◽

Lupus Erythematosus ◽

Skin Lesions ◽

Epidermal Keratinocytes ◽

Systemic Lupus ◽

Tnf Α ◽

Ifn Γ

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In vitro C3 mRNA Expression in Pemphigus Vulgaris: Complement Activation is Increased by IL-1α and TNF-α

Journal of Cutaneous Medicine and Surgery ◽

10.1177/120347549900300306 ◽

1999 ◽

Vol 3 (3) ◽

pp. 140-144 ◽

Cited By ~ 20

Author(s):

Claudio Feliciani ◽

Paola Toto ◽

Paolo Amerio ◽

Pierluigi Amerio

Keyword(s):

Mrna Expression ◽

Complement Activation ◽

In Vitro Study ◽

Peak Level ◽

Pemphigus Vulgaris ◽

Epidermal Keratinocytes ◽

Human Epidermal Keratinocytes ◽

Tnf Α

Background: Pemphigus vulgaris (PV) is a potentially life-threatening disease, characterized immunohistologically by IgG deposits and complement activation on the surface of keratinocytes. Complement activation has been implicated in the pathogenesis with C3 deposits in about 90% of patients. Objective: In order to further elucidate the role of complement in PV and to define which cytokines play a role in C3 mRNA expression, we performed an in vitro study in human keratinocytes. Methods: Normal human epidermal keratinocytes (NHuK) were incubated with PV serum and C3 mRNA was measured. We previously had shown that IL-1α and TNF-α are expressed in PV in vivo and in vitro. Since cytokines are able to modulate complement activation, mRNA expression was evaluated in a similar experiment after pretreatment using antibodies against IL-1α and TNF-α. Results: Incubation of NHuK with PV sera caused their detachment from the plates after 20–30 minutes with a complete acantholysis within 12 hours. An early C3 mRNA expression was seen after 30 minutes with a peak level after 1 hour. Blocking studies, using antibodies against human IL-1α and TNF-α in NHuK together with PV-IgG, showed reduction of in vitro induced acantholysis and inhibition of C3 mRNA expression. Conclusions: This study supports the hypothesis that complement C3 is important in PV acantholysis and that complement activation is increased by IL-1α and TNF-α.

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Comparative Study of the Effect of 5,6-Dihydroergosterol and 3-epi-5,6-dihydroergosterol on Chemokine Expression in Human Keratinocytes

Molecules ◽

10.3390/molecules25030522 ◽

2020 ◽

Vol 25 (3) ◽

pp. 522

Author(s):

Ye Seul Park ◽

Hye Jin Moon ◽

Kwang Hyun Ahn ◽

Tae Hoon Lee ◽

Hakwon Kim

Keyword(s):

Mrna Expression ◽

Inhibitory Activity ◽

Human Keratinocytes ◽

Expression Levels ◽

Chemokine Expression ◽

Tnf Α ◽

New Type ◽

Synthetic Derivative ◽

Ifn Γ ◽

Mrna Expression Levels

5,6-Dihydroergosterol-glucose is an organic synthetic derivative of spinasterol-glucose, which has potent anti-inflammatory activity. We previously synthesized alpha and beta anomers of DHE-glycosides and compared their inhibitory activity on CCL17 and CCL22 mRNA expression induced by TNF-α/IFN-γ in activated HaCaTs. Recently, we synthesized a new type of DHE-glycosides, 3-epi-5,6-dihydroergosterol(3-epi-DHE)-glycosides, and compared its inhibitory activity on mRNA expression levels of CCL17 and CCL22 in TNF-α/IFN-γ-induced HaCaT. DHE-Xly did not affect TNF-α/IFN-γ induced CCL17 and CCL22 mRNA expression in HaCaTs, however, 3-epi-DHE-Xly strongly inhibited TNF-α/IFN-γ induced CCL17 and CCL22 mRNA expression levels in human keratinocytes. These results provide important clues for development of chronic dermatitis treatment via inhibition of chemokine expression using DHE derivatives.

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The Immunomodulatory Effects of Phellodendri Cortex Polysaccharides on Cyclophosphamide-Induced Immunosuppression in Mice

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2021/3027708 ◽

2021 ◽

Vol 2021 ◽

pp. 1-9

Author(s):

Yi Cheng ◽

Lu Liu ◽

Simei Mo ◽

Jianxiong Gao ◽

Hongjian Zhang ◽

...

Keyword(s):

Mrna Levels ◽

Immunomodulatory Effects ◽

Control Groups ◽

Tnf Α ◽

Cyclophosphamide Treatment ◽

Ifn Γ ◽

Phellodendri Cortex ◽

Spleen Lymphocytes ◽

Immunosuppressed Mice

Cyclophosphamide is a commonly used anticancer drug, and immunosuppression is one of the most common side effects. How to recover the immunological function is important for cyclophosphamide-treated patients. In the present study, Phellodendri Cortex polysaccharides (CPP) could enhance the proliferation of mouse spleen lymphocytes in vitro. The immunoregulatory function of CPP was then investigated in cyclophosphamide-induced immunosuppressed mice. In CPP-treated groups, mice were orally treated with CPP at doses of 1, 0.5, and 0.25 g/kg bodyweight from 1 to 11 d, respectively. The cyclophosphamide was administrated in CPP and cyclophosphamide groups from 12 to 14 d. In the cyclophosphamide and normal control groups, the mice received equal volume of saline from 1 to 14 d. The results showed that CPP (1 g/kg) could significantly increase the bodyweight of mice, even during cyclophosphamide treatment. The organ coefficients of the spleen and thymus were recovered by CPP treatment. CPP upregulated the contents of cytokines (IL-2, IL-6, IFN-γ, and TNF-α) in serum, which were downregulated by cyclophosphamide. The mRNA levels of these cytokines were also elevated by CPP treatment in the spleen. Cyclophosphamide upregulated the expressions of NF-κB p65, TLR4, and MyD88, suggesting that the NF-κB signaling pathway was activated by cyclophosphamide. After CPP treatment, it was recovered to normal level. These results indicated that CPP alleviated the cyclophosphamide-induced immunosuppression.

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Alterations of ultrastructure and elemental composition in cultured human epidermal keratinocytes after treatment with nitrofurazone and silver sulfadiazine

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100127803 ◽

1987 ◽

Vol 45 ◽

pp. 676-677

Author(s):

A. R. Crooker ◽

M. C. Myers ◽

T. L. Beard ◽

E. S. Graham

Keyword(s):

Electron Microscopy ◽

Elemental Composition ◽

Pyrolytic Carbon ◽

Human Keratinocytes ◽

Silver Sulfadiazine ◽

Epidermal Keratinocytes ◽

Human Epidermal Keratinocytes ◽

Culture Systems ◽

Cytotoxicity Endpoints

Cell culture systems have become increasingly popular as a means of screening toxic agents and studying toxic mechanisms of drugs and other chemicals at the cellular and subcellular levels. These in vitro tests can be conducted rapidly in a broad range of relevant mammalian culture systems; a variety of biological and biochemical cytotoxicity endpoints can be examined. The following study utilized human keratinocytes to evaluate the relative cytotoxicities of nitrofurazone (NF) and silver sulfadiazine (SS), the active ingredients of FURACIN(R) Topical Cream and SILVADENE(R) Cream, respectively. These compounds are anti-infectives used in the treatment of burn patients. Cell ultrastructure and elemental composition were utilized as cytotoxicity endpoints.Normal Human Epidermal Keratinocytes (HK) were prepared from the EpiPackTM culture system (Clonetics Corporation, Boulder, CO). For scanning electron microscopy (SEM) and transmission electron microscopy (TEM), cells were seeded on sterile 35 mm Falcon plastic dishes; for elemental microanalysis, cells were plated on polished pyrolytic carbon discs (E. Fullam, Latham, NY) placed in the culture dishes.

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