Th2 Responses Elicited by Nasal Epithelial Cells Exposed to House Dust Mite Extract

Objectives

Respiratory epithelial cells are the first site of interaction of allergens with the immune system. The aim of this study was to examine the effect of epithelial cells, which were stimulated with house dust mite (HDM) extracts, on the immune response of peripheral blood mononuclear cells (PBMCs).

Methods

Primary nasal polyp epithelial cells were exposed to dermatophagoides pteronyssinus and dermatophagoides farina for 48 hr, and then the supernatant and cells were collected. After stimulation with HDM extract, the epithelial cells were co-cultured with PBMCs for 72 hr and then the supernatant was collected. We measured the interleukin (IL)-8 and granulocyte-macrophage colony stimulating factor to determine the activation of the epithelial cells. The tumor necrosis factor (TNF)-α, IL-5 and interferon-γ were measured to evaluate the interaction between the epithelial cells and the PBMCs. The mRNA expression of intercellular adhesion molecule 1 (ICAM-1) was assessed using the anti-ICAM-1 antibody.

Results

The HDM extracts activated the nasal epithelial cells and enhanced the expression of ICAM-1 mRNA and cell membrane ICAM-1. When the activated epithelial cells were co-cultured with PBMCs, the PBMCs produced lager amounts of TNF-α and IL-5. However the cytokine production was not inhibited by pretreatment with ICAM-1 antibody.

Conclusion

HDM allergens induce allergic inflammation by activating nasal epithelial cells, yet the interaction of the epitheila cells and the PBMCs may not be associated with an ICAM-1 medicated mechanism.     

Globus Pallidus Interna Deep Brain Stimulation for Chorea-Acanthocytosis

Chorea-acanthocytosis (ChAc) is a rare hereditary disorder characterized by involuntary choreiform movements and erythrocytic acanthocytosis. Pharmacotherapy for control of involuntary movements has generally been of limited benefit. Deep brain stimulation (DBS) has recently been used for treatment of some refractory cases of ChAc. We report here on the effect of bilateral high-frequency DBS of globus pallidus interna in a patient with ChAc.     

Vanillic acid inhibits inflammatory mediators by suppressing NF-κB in lipopolysaccharide-stimulated mouse peritoneal macrophages

Vanillic acid is a benzoic acid derivative that is used as a flavoring agent. It is an oxidized form of vanillin. At present, the mechanisms by which vanillic acid exerts its anti-inflammatory effects are incompletely understood. In this study, we attempted to determine the effects of vanillic acid on lipopolysaccharide (LPS)-induced inflammatory responses in mouse peritoneal macrophages. Our findings indicate that vanillic acid inhibits LPS-induced production of tumor necrosis factor (TNF)-α and interleukin (IL)-6. During the inflammatory process, the levels of cyclooxygenase (COX)-2 and nitric oxide (NO) increased in mouse peritoneal macrophages, but vanillic acid suppressed both the enhanced levels of COX-2 and the production of prostaglandin E(2) and NO. Moreover, vanillic acid suppressed the activation of nuclear factor-kappa B (NF-κB) and caspase-1. These results provide novel insights into the pharmacological actions of vanillic acid and are indicative of the potential use of this molecule in the treatment of inflammatory diseases.     

Insamhodo-tang, a traditional Korean medicine, regulates mast cell-mediated allergic inflammation in vivo and in vitro

Aim of the study

Insamhodo-tang (IHT) has traditionally been used in Korea to treat a variety of diseases, including chronic cough, tuberculosis, and chronic bronchitis. However, the anti-allergic and anti-inflammatory effects of IHT and its molecular mechanisms have yet to be clearly elucidated. In this study, we attempted to evaluate the effects of IHT on mast cell-mediated allergy inflammation in vitro and in vivo.

Materials and methods

We investigated to ascertain the pharmacological effects of IHT on both compound 48/80-induced and 2,4-dinitrofluorobenzene (DNFB)-induced allergic reactions under in vivo conditions. Additionally, to find a possible explanation for the anti-inflammatory mechanisms of IHT, we evaluated the regulatory effects of IHT on the level of inflammatory mediators in phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI)-stimulated human mast cells (HMC-1).

Results

The finding of this study demonstrated that IHT reduced compound 48/80-induced systemic anaphylactic shock, DNFB-induced dermatitis, and ear swelling responses in mice. Additionally, IHT inhibited the production of interleukin (IL)-6, IL-8, and TNF-α, as well as the activation of nuclear factor-κB and caspase-1 in PMACI-stimulated HMC-1.

Conclusion

Collectively, the findings of this study provide us with a novel insight into the pharmacological actions of IHT as a potential molecule for use in the treatment of allergic inflammation diseases.     

Use of scopoletin to inhibit the production of inflammatory cytokines through inhibition of the IkappaB/NF-kappaB signal cascade in the human mast cell line HMC-1

Scopoletin (6-methoxy-7-hydroxycoumarin) is a coumarin compound and a pharmacologically active agent that has been isolated from several plant species. However, as yet there is no clear explanation of how scopoletin affects the production of inflammatory cytokine. We therefore used cells from the human mast cell line (HMC-1) to investigate this effect. Scopoletin significantly and dose-dependently inhibits the way in which phorbol 12-myristate 13-acetate (PMA) plus A23187 induces the production of inflammatory cytokines such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-8 (P<0.05). The maximal rates at which scopoletin (0.2 mM) inhibited the production of TNF-alpha, IL-6, and IL-8 were 41.6%+/-4.2%, 71.9%+/-2.5%, and 43.0%+/-5.7%, respectively. In activated HMC-1 cells, the expression level of nuclear factor (NF)-kappaB/Rel A protein was increased in the nucleus whereas the level of NF-kappaB/Rel A in nucleus was decreased by treatment with scopoletin. Scopoletin decreased PMA plus A23187-induced luciferase activity. Scopoletin also inhibits IkappaBalpha phosphorylation and degradation in cytoplasm. These results indicate that scopoletin has a potential regulatory effect on inflammatory reactions that are mediated by mast cells.     

Induction of apoptosis by KI0477959 through activation of caspase-3 in human leukemia cell line, HL-60 cells

KI0477959 (Herbkines) has been used for the purpose of development of physical strength in wasting diseases, like cancer. In the present study, apoptosis-inducing activities of butanol fraction of KI0477959 were studied in human leukemia cell line, HL-60 cells. KI0477959 increased cytotoxicity but had less effect on human peripheral blood mononuclear cells. KI0477959-induced apoptosis was accompanied by activation of caspase-3 and specific proteolytic cleavage of poly-ADP-ribose polymerase. Increased apoptosis was reduced by treatment with p38 and extracellular signal-regulated protein kinase (ERK) inhibitors. These results suggest that KI0477959 induces apoptosis through activation of caspase-3, p38, and ERK in HL-60 cells.     

Transgenic Panax ginseng inhibits the production of TNF-alpha, IL-6, and IL-8 as well as COX-2 expression in human mast cells

The most well-known medicinal plant, Panax ginseng (P. ginseng), contains various phytosterols and bioactive triterpene saponins (ginsenosides). Squalene synthase is a key regulatory enzyme for triterpene biosynthesis and overexpression of the squalene synthase confers the hyper-production of triterpene saponins to form transgenic ginseng. In this study, we have investigated whether and how transgenic P. ginseng modulates an inflammatory reaction in a stimulated human mast cell line, HMC-1. It was found that transgenic P. ginseng inhibited the production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-8, and the expression of cyclooxygenase-2 in phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187 (PMACI)-stimulated HMC-1. Additionally, we have shown that transgenic P. ginseng suppressed the intracellular calcium level induced by PMACI. These results provide new insights into the pharmacological actions of transgenic P. ginseng as a potential molecule for use in therapy in mast cell-mediated inflammatory diseases.     

Levocetirizine inhibits rhinovirus-induced bacterial adhesion to nasal epithelial cells through down-regulation of cell adhesion molecules

BackgroundThe first critical step for bacterial infection is attachment of bacteria to the cell adhesion molecules of epithelial cells. The rhinovirus (RV)-induced increased expression of cell adhesion molecules including fibronectin (Fn) and carcinoembryonic antigen–related cell adhesion molecules (CEACAMs) is closely related to the activation of nuclear factor-kappa B (NF-κB). Recent studies have demonstrated that Levocetirizine (LCT) has anti-inflammatory properties that are mediated by inhibitory effects on NF-κB in addition to classic antihistaminic effects.ObjectiveTo investigate the inhibitory effects of LCT on the RV-induced expression of Fn and CEACAMs in human nasal epithelial cells (HNECs) and identified the effects of LCT on secondary Staphylococcus aureus and Haemophilus influenzae adhesion to RV-infected HNECs.MethodsPrimary HNECs obtained from inferior turbinate mucosa were pretreated with 50 nM LCT 24 hours before RV-16 infection and for 48 hours thereafter. The expression levels of Fn and CEACAMs were assayed by real-time polymerase chain reaction (PCR) and Western blotting. Bacterial adhesion to cells was assessed by confocal microscopy.ResultsFibronectin and CEACAM messenger RNA (mRNA) and protein levels in HNECs were significantly increased by RV-16 infection. Levocetirizine significantly reduced these increases in mRNA levels and protein expression of Fn and CEACAMs. Confocal microscopy showed that treatment with LCT significantly reduced the adhesion levels of S aureus and H influenza in RV-infected HNECs compared with RV-infected, untreated HNECs.ConclusionThese findings suggest that LCT inhibits the expression of Fn and CEACAMs and has the potential to prevent secondary bacterial infections in RV-infected HNECs by interfering with bacterial adhesion.     

The Effect of Nano-Silver on Allergic Rhinitis Model in Mice

Objectives

Silver has long been known as a strong antimicrobial and disinfectant. Several types of nano-silver coated products have been developed. However, the antimicrobial and disinfectant characteristics of nano-silver have not been well studied. The aim of this study was to investigate the effect of nano-silver on allergic inflammation in a mouse model.

Methods

Female BALB/C mice were sensitized by intraperitoneal injection of ovalbumin (OVA) and aluminium hydroxide on days 0, 7, 14, and 21. Mice were challenged with intranasal instillation of OVA. Nano-silver was also administered nasally prior to intranasal instillation of OVA. Severity of allergic rhinitis was assessed according to nasal symptoms, serum OVA-specific IgE level, interleukin (IL)-4, IL-10, and interferon (INF)-γ levels in nasal lavage fluid. Hematoxylin-eosin stain and periodic acid-Schiff stain were performed for evaluation of histological change.

Results

Nano-silver attenuated manifestation of nasal symptoms in sensitized mice and inhibited production of OVA-specific IgE, IL-4, and IL-10, however, it had no effect on INF-γ level. In addition, the degree of inflammatory cell infiltration and goblet cell hyperplasia was attenuated by nano-silver.

Conclusion

These results suggest that nano-silver may effectively reduce allergic inflammation in a mouse model of allergic rhinitis. Through its properties as an anti-inflammatory agent, nano-silver may be a useful therapeutic strategy.