Microglial activation and inflammation are associated with progressive neuronal apoptosis in neurodegenerative human brain disorders. We sought to investigate molecular signaling mechanisms that govern activation of microglia in apoptotic neuronal degeneration. We report here that the active form of matrix metalloproteinase-3 (MMP-3) was released into the serum-deprived media (

Allogenic myoblast transplantation (AMT) is under investigation for treatment of severe genetic myopathies. Data regarding the role of cyclosporine (CsA) and FK-506 in AMT have shown that CsA is less effective than FK-506. For this study, we investigated mechanisms of CsA toxicity during AMT and showed that a high level of reactive oxygen species (ROS) generated by CsA, mediated partly by inhibition of the peptidylprolyl-cis-trans-isomerase (PPIase)-like activity of cyclophilin A (CypA), blocked differentiation and induced apoptosis at an early stage of muscle differentiation. Inhibition of the PPIase-like activity of CypA alone also blocked muscle differentiation. However, CsA toxicity did not depend on the inhibition of calcineurin activity during muscle differentiation. Together, these data suggest that CsA-mediated inhibition of the PPIase-like activity of CypA and the high level of ROS generation contributed to the low efficacy of CsA in AMT. In addition, we showed that a reduction of oxidative stress protected cells from CsA-induced apoptosis, and myoblasts that had survived after preexposure to CsA not only proliferated and differentiated reversibly but also gained resistance to subsequent CsA exposure. Thus, administration of antioxidants or overexpression of CypA either exogenously or endogenously during CsA treatment has the potential to improve the success of this treatment in AMT.

Atopic dermatitis (AD) is one of the most common skin diseases, and is characterized by a chronic and relapsing inflammatory dermatitis with immunological disturbance, e.g., hyper-production of total and specific IgE. Patients with AD usually have a personal or familial history of other atopic diseases.1) Population studies suggest that in most countries AD affects 10-20% of all children at some time during childhood.2) Moreover, AD is a multifactorial disease, and its pathogenesis and etiology are not fully understood. 1)In acute AD lesions, cells expressing T helper 2 (Th2) cytokines such as interleukin-4 (IL-4) and IL-5, are significantly increased, whereas delayed-type allergic reaction and Th1 immune response characterize the pathogenesis of chronic AD. 3)Delayed-type hypersensitivity (DTH) reactions, such as, chemical contact allergy, are usually regarded as cell-mediated immune responses, which are frequently attributed to T helper (Th) 1-type cells. 4) Moreover, dinitrofluorobenzene (DNFB) induced contact hypersensitivity is a T cell-mediated inflammatory skin reaction that is thought to be associated with the activation of type1 helper T cells. 5)NC/Nga mice are the most extensively studied animal model of AD. The NC/Nga strain originates from Japanese fancy mice, and was established as an inbred strain in 1957. NC/Nga mice have also been reported to develop AD-like eczematous skin lesions when kept in an air-uncontrolled conventional housing but not when maintained under specific pathogen-free (SPF) conditions.2) Clinical symptoms begin with itching, erythema, hemorrhage, scaling, dryness, and alopecia at age 8 weeks. Given the resemblance between these immunological alterations in NC/Nga mice and AD in humans, models based on these mice are of importance for the understanding and treatment of AD.2)The roots of Astragalus membranaceus (AM) are one of the most popular health-promoting herbs in East Asia, and have been used for more than 2000 years.6) The main constituents of AM roots are polysaccharides, saponins, flavonoids, amino acids, and trace elements. 7) Recently AM is reported that it enhanced the levels of IL-4 but reduced level of IFN-g secretion in TCR (T cell receptor) stimulated T cells.8) Additionally, AM in combination with other herbs, Hochu-ekki-to, was used to treat allergic diseases. 9-11)Hochu-ekki-to also known to suppressed dermatitis and serum IgE levels in NC/Nga mice. 9) However, Hochu-ekki-to contain many different constituents, it is difficult to identify the active components. Therefore, in this study, we examined whether AM suppresses AD-like skin lesions in NC/Nga mice. MATERIALS AND METHODSMice Female NC/Nga mice, 8 weeks of age, were obtained from Orient Korea (Gyeonggi, Korea) and maintained under SPF conditions. Animals were housed in an air conditioned animal room at 25Ϯ1°C and a relative humidity (RH) of 40Ϯ5%, and fed a laboratory diet and water. Experiments conducted in accord with the guidelines issued by the Ethical Committee for Animal Welfare at KyungHee Univers...

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Consumption of omega-3 polyunsaturated fatty acid, particularly eicosapentaenoic acid (EPA), is associated with a significant reduction in the risk of developing cardiovascular disease. The aim of this study was to investigate whether heme oxygenase-1 (HO-1) induction contributes to the cytoprotective effects of EPA in endothelial cells threatened with oxidative damage. In this study, we investigated the effect of EPA on the induction of HO-1 by NF-E2-related factor 2 (Nrf2) in human umbilical vein endothelial cells. In cells treated with low concentrations of EPA (10-25 μM), HO-1 expression increased in a time- and concentration-dependent manner. Additionally, EPA treatment increased Nrf2 nuclear translocation and antioxidant response element activity, leading to the upregulation of HO-1 expression. Furthermore, treatment with EPA reduced hydrogen peroxide (H(2)O(2))-induced cell death. The reduction in cell death was reversed by treatment with zinc protoporphyrin, an inhibitor of HO-1, indicating that HO-1 contributed to the protective effect of EPA. These data suggest that EPA protects against H(2)O(2)-induced oxidative stress in endothelial cells by activating Nrf2 and inducting HO-1 expression.

Melatonin possesses a number of important biologic activities including oncostatic, anti-oxidant, and immunostimulatory actions. This study was designed to assess the effects of melatonin on inflammation-related gene expression in lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMCs), using CombiMatrix 2K Human Inflammation chip. After pretreatment with melatonin (100 microm) for 4 hr, cells were incubated with LPS (1 microg/mL) for 24 hr. We compared gene expression profiles between LPS-treated, melatonin-treated, LSP/melatonin-treated, and control groups. LPS induced the upregulation of 95 genes, compared with controls. Melatonin pretreatment in LPS-stimulated PBMCs suppressed the expression of 23 genes more than twofold. Interestingly, melatonin showed a suppressive effect on the expression of CC chemokine subfamily genes, including CCL2/MCP1, CCL3/MIP1 alpha, CCL4/MIP1 beta, CCL5/RANTES, CCL8/MCP2, CCL20/MDC, and CCL22/MIP3 alpha, in LPS-stimulated PBMCs. This result was confirmed by reverse transcriptase polymerase chain reaction. Among the CC chemokine subfamily genes, particularly, the expression of CCL2 and CCL5 was markedly downregulated by melatonin in LPS-stimulated PBMCs. The secretion levels of CCL2 and CCL5 were measured using enzyme-linked immunosorbent assay. Stimulation of PBMCs by LPS induced the secretion of CCL2 (2334.3 +/- 161.4 pg/mL, mean +/- S.E.M.), whereas melatonin pretreatment (153.0 +/- 3.8 pg/mL) inhibited the LPS-induced secretion of CCL2. Melatonin pretreatment (2696.2 +/- 385.3 pg/mL) also inhibited the LPS-induced secretion of CCL5 (4679.6 +/- 107.5 pg/mL). Taken together, these results suggest that melatonin may have a suppressive effect on LPS-induced expression of CC chemokine genes, especially CCL2 and CCL5, which may explain its beneficial effects in the treatment of various inflammatory conditions.

Acute pancreatitis (AP) is a clinical entity that is believed to have intracellular activation of digestive enzymes and autodigestion of the pancreas as its central pathophysiologic cause. The noninfectious destruction of pancreatic parenchyma quickly induces an inflammatory reaction at the site of injury. Histologically, AP is characterized by interstitial edema, vacuolization, inflammation and acinar cell necrosis. The diagnosis of AP is based on pancreatic edema index (pancreas weight/body weight) and pancreatic serum enzymes (pancreatic amylase, lipase, immunoreactive trypsin or elastase. 1,2)The induction of the heat shock response enhances the ability of the cells to overcome the effects of the stress.3) The heat shock proteins (HSPs) are involved in the synthesis, degradation, folding, transport, and translocation of proteins. 4,5) HSPs have been classified into six families according to their molecular mass. It is well known that the increased expression of HSPs have protective effects against caerulein-induced pancreatitis in rats or against choline-deficient ethionine-supplemented diet pancreatitis model. [6][7][8] Others reported that the pre-induction of HSP expression has a protective effect against caerulein-induced pancreatitis in mice. Many diseases result in increased levels of HSPs. But caerulein or CCK-induced pancreatitis reduces the levels of pancreatic HSPs. 9,10) Cytokines are important immunoregulatory mediators. Their contribution to the pathogenesis of acute and chronic gastroenterological disorders is obvious. Increased expression of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor (TNF)-a can be detected in inflammatory bowel disease. Pro-inflammatory cytokines such as IL-1, TNF-a, and IL-6 are elevated during AP and are considered to be involved in the pathogenesis of pancreatitis-associated multiple organ dysfunction.11) Elevated serum levels of these cytokines have been demonstrated clinically as well as experimentally in different animal models of AP. 12)Among the neurohormonal regulators, Cholecystokinin (CCK) is a well known gastrointestinal hormone and neural agonist to induce the release of pancreatic digestive enzymes 13) . At supramaximal doses (doses greater than those that cause maximal secretion of digestive enzymes by the pancreatic acinar cell), CCK is able to cause pancreatic responses. 14,15) Statins, the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, are the most potent lipidlowering agents currently available, and are being prescribed in the treatment of hyperlipidemia. 16,17) Statins are also effective for the reduction of vascular inflammation. Thus, we set out to investigate the effects of statin as anti-inflammatory agent on AP. MATERIALS AND METHODSReagents Avidin-peroxidase and 2Ј-AZINO-bis (3-ethylbenzithiazoline-6-sulfonic acid) tablets substrate and CCK-8 were purchased from Sigma (St. Louis, MO, U.S.A.). Anti-HSP60 antibodies were purchased from Stressgen (British Columbia, Canada). Anti-rat IL-6, TNF-a and IL-1b ant...

Crotonaldehyde (CRA) is an α,β-unsaturated aldehyde that is genotoxic, carcinogenic, and mutagenic, and forms 1,N(2)-propanodeoxyguanosine. Humans are exposed to CRA in work places, and from tobacco smoke, environmental pollution, food, and beverages. In addition, CRA is a product of endogenous lipid peroxidation, arising as a consequence of oxidative stress and electrophilic stress, which may be related to development of cardiovascular diseases. In this study, we used 24 k whole human DNA chips for identification of gene expression profiles by treatment of CRA in human umbilical vein endothelial cells (HUVECs). Among the 24,000 genes of cDNA microarrays, we identified up-and down-regulated genes that changed by more than 1.8 and 2 fold by CRA. Our data showed that CRA changed gene expression patterns at the genome-wide transcriptional level in HUVECs. And several genes showed association with cardiovascular diseases.