Areca nut extracts increased the expression of cyclooxygenase-2, prostaglandin E2 and interleukin-1α in human immune cells via oxidative stress.

BACKGROUND AND OBJECTIVES: Areca nut has been identified as a carcinogen. Inflammation reveals a strong link with tumourigenesis. The aim of this study was to investigate the effects of areca nut on the expression of the key pro-inflammatory mediators involved in malignancy, cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2), interleukin (IL)-1α and nuclear factor-κB (NF-κB), by human immune cells. The role of oxidative stress was also examined. MATERIALS AND METHODS: Human peripheral blood mononuclear cells (PBMCs) were treated with extracts of ripe areca nut (rANE) or tender areca nut (tANE). Expression of pro-inflammatory mediators was assayed using Western blotting, reverse transcription-polymerase chain reaction, competitive enzyme immunoassay or enzyme-linked immunosorbent assay (ELISA). Activity of NF-κB was evaluated using an ELISA-based method. RESULTS: Both rANE and tANE enhanced the expression of COX-2, PGE2 and IL-1α by PBMCs. The secretion of PGE2 was induced by rANE (≤20-40µgml(-1)) and tANE (≤160µgml(-1)) significantly in a dose- and time-dependent manner. However, the above enhancing effects of ANEs could be attenuated by antioxidants. ANEs also increased the nuclear expression of the redox-sensitive factor NF-κB. CONCLUSIONS: The results demonstrate that ANEs induced the expression of pro-inflammatory mediators mainly through the induction of oxidative stress and implicate the possibility of using antioxidants for disease prevention.

Lipid bodies in inflammatory cells: structure, function, and current imaging techniques.

Lipid bodies (LBs), also known as lipid droplets, have increasingly been recognized as functionally active organelles linked to diverse biological functions and human diseases. These organelles are actively formed in vivo within cells from the immune system, such as macrophages, neutrophils, and eosinophils, in response to different inflammatory conditions and are sites for synthesis and storage of inflammatory mediators. In this review, the authors discuss structural and functional aspects of LBs and current imaging techniques to visualize these organelles in cells engaged in inflammatory processes, including infectious diseases. The dynamic morphological aspects of LBs in leukocytes as inducible, newly formable organelles, elicitable in response to stimuli that lead to cellular activation, contribute to the evolving understanding of LBs as organelles that are critical regulators of different inflammatory diseases, key markers of leukocyte activation, and attractive targets for novel anti-inflammatory therapies.

Subcellular localization of Mitf in monocytic cells.

Microphthalmia-associated transcription factor (Mitf) is a transcription factor that plays an important role in regulating the development of several cell lineages. The subcellular localization of Mitf is dynamic and is associated with its transcription activity. In this study, we examined factors that affect its subcellular localization in cells derived from the monocytic lineage since Mitf is present abundantly in these cells. We identified a domain encoded by Mitf exon 1B1b to be important for Mitf to commute between the cytoplasm and the nucleus. Deletion of this domain disrupts the shuttling of Mitf to the cytoplasm and results in its retention in the nucleus. M-CSF and RANKL both induce nuclear translocation of Mitf. We showed that Mitf nuclear transport is greatly influenced by ratio of M-CSF/Mitf protein expression. In addition, cell attachment to a solid surface also is needed for the nuclear transport of Mitf.

Roles and origins of leukocyte lipid bodies: proteomic and ultrastructural studies.

Lipid bodies (LBs), multifunctional organelles present in most eukaryotic cells, are sites of eicosanoid formation in leukocytes; but little is known about the composition of leukocyte LBs or the biogenesis and internal structures of LBs from mammalian cells. Proteomic analyses of LBs purified from human monocytic U937 cells detected, common to LBs in other cells, proteins involved in cholesterol and triglyceride metabolism, Rab GTPases, and many membrane and endoplasmic reticulum (ER)-associated proteins. Newly lipid body (LB)-associated proteins included MRP-14, potentially involved in arachidonate transport, and ribosomal subunit proteins and translation regulatory proteins. Ultrastructurally, in U937 cells as well as human neutrophils and eosinophils, ribosomes are attached to and distributed within LBs, and LBs contain extensive ER-like membranes. The presence of ribosomes, ER-like membranes and many membrane-associated and ER luminal proteins within LBs, supports a new model by which enveloped ER-membranes and domains form LBs and indicates that LBs may be sites of protein synthesis.

Enhancing effects of areca nut extracts on the production of interleukin-6 and interleukin-8 by peripheral blood mononuclear cells.

BACKGROUND: The habit of chewing areca quid (AQ) has been implicated in oral pathogenesis, including periodontal disease. Little is understood about the roles of AQ in the cytokine secretion by immune cells. The study examined the effects of areca nut, the major ingredient of AQ, on the production of interleukin (IL)-6 and IL-8 by peripheral blood mononuclear cells (PBMC), the immunocompetent cells. The possible role of oxidative stress of areca nut was also examined. METHODS: Extracts of ripe areca nut (rANE) and tender areca nut (tANE) were examined for their cytotoxic effects on human PBMC using the trypan blue exclusion test. The production of IL-6 and IL-8 by ANE-treated PBMC was analyzed using enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction. Effects of an antioxidant, pyrrolidine dithiocarbamate (PDTC), on ANE-induced cytokine secretion were also studied. RESULTS: At the experimental conditions, 20 micro g/ml rANE decreased cell viability significantly, whereas no significant effect of tANE (< or =80 micro g/ml) was observed. Both rANE (< or =20 micro g/ml) and tANE (< or =160 micro g/ml) significantly increased the secretion of IL-6 and IL-8 by PBMC in a dose- and time-dependent manner. The altered mRNA expression of IL-6 by rANE and tANE was also observed. Moreover, the stimulating effects of rANE on cytokine expression in PBMC could be attenuated by PDTC, suggesting that the oxidative stress of rANE may play a role. CONCLUSIONS: Markedly enhancing effects of ANE on PBMC-released inflammatory cytokines might cause a sustained cytokine-rich inflammatory milieu in oral cavity of AQ chewers. These excessive cytokines from ANE-treated immune cells may impair periodontal health.