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Objective Inflammation is crucial to limiting vascular disease. Previously we reported that acrolein, a known toxin in tobacco smoke, might play an important role in the progression of atherosclerosis via an inflammatory response involving cyclooxygenase-2 (COX-2) and prostaglandin production in human umbilical vein endothelial cells (HUVECs). Curcumin has been known to improve vascular function and have anti-inflammatory properties. In this study, we investigated whether curcumin prevents the induction of inflammatory response caused by acrolein. Methods Anti-inflammatory effects of curcumin were examined in acrolein-stimulated HUVECs. Induction of proteins, mRNA, prostaglandin and reactive oxygen species (ROS) were measured using immunoblot analysis, real-time reverse-transcription polymerase chain reaction, enzyme-linked immunosorbent assay and flow cytometry, respectively. Results Curcumin attenuates inflammatory response via inhibition of COX-2 expression and prostaglandin production in acrolein-induced human endothelial cells. This inhibition by curcumin results in the abolition of phosphorylation of protein kinase C, p38 mitogen-activated protein kinase, and cAMP response element-binding protein. Furthermore, curcumin suppresses the production of ROS and endoplasmic reticulum stress via phosphorylation of eukaryotic initiation factor-2α caused by acrolein. Conclusion These results suggest that curcumin might be a useful agent against endothelial dysfunction caused by acrolein-induced inflammatory response.

Biodegradable polymers, such as poly(L-lactide) (PLLA), are very useful in many biomedical applications. However, their degradation by-products have been much of a concern as they are the sources of inflammatory reactions in the body. In this work, we suggest a novel composite system composed of PLLA and oligolactide-grafted magnesium hydroxide (Mg-OLA) that can overcome drawbacks caused by poor mechanical properties and inflammatory response of PLLA for biomedical applications. Mg-OLAs were synthesized by ring opening polymerization and the structure, morphology, pH change, thermal, and mechanical properties were analyzed using FTIR, SEM, pH meter, TGA, and UTM. In particular, the tensile strength and modulus of PLLA/Mg80-OLA20 (0–20 wt%) were higher than those of PLLA/magnesium hydroxide. The PLLA/Mg80-OLA20 composite was also very effective in neutralizing the acidic environment caused by the degradable by-product of the PLLA matrix. In vitro cell viability and the expression levels of COX-2 and IL-6 proteins in the PLLA composites were also evaluated. Cell viability increased to around 100% with increasing the amount of Mg80-OLA20 from 0 to 20 wt%. The expression levels of IL-6 and COX-2 were reduced dramatically when increasing the proportion of Mg80-OLA20 from 0 to 50 wt%. As a result, the incorporation of Mg-OLAs into the PLLA matrix could reinforce the mechanical properties as well as reduce the inflammatory response of the hybrid PLLA. Therefore, this hybrid composite system blending oligomer-grafted magnesium hydroxide in biodegradable polymers would be a promising strategy for avoiding current fatal problems in biomedical applications.

Diabetic nephropathy (DN) is one of the complications of diabetes mellitus (DM) and the worldwide prevalence of DN has continued to increase. DN causes cellular stress and inflammation, which induces death of renal cells and dysfunction of blood vessels. MicroRNAs (miRNAs) are small interfering RNAs that negatively regulate mRNA gene expression. Various human diseases are attributed to aberrant miRNAs expression, and miRNAs levels are used as markers for diagnosis and classification of diseases. However, the profile of miRNAs in DN mouse have not been fully explained, we analyzed miRNAs expression in the kidney of diabetic mice. C57BL6 male mice were injected with streptozotocin (50 mg/kg body weight) for five consecutive days to induce DM. Mice were sacrificed after 16 weeks and RNAs were extracted from the kidneys and analyzed via microarray profiling. Our results confirmed that 137 miRNAs expression was altered in DM. These miRNAs showed pairwise correlations with 48 mRNAs in DM kidney. Furthermore, Differential expressed miRNAs were characterized by Gene Ontology (GO) enrichment analysis and signaling pathway using DAVID. Our findings may provide that changes in miRNA expression in mice with diabetic nephropathy influences disease progression and might provide insights into the molecular mechanisms underlying DN.

Background Adiponectin is an adipocyte-derived cytokine that circulates as a full-length protein and a fragment containing the globular domain of adiponectin (gAd). A recent study has reported the antimelanogenic effects of full-length adiponectin. Objectives To examine the involvement of gAd in melanogenesis and its mechanisms of action. Methods The effects of gAd on melanogenesis and its mechanisms of action were investigated in human epidermal melanocytes and reconstructed epidermis, including melanin content, cellular tyrosinase activity, cyclic adenosine monophosphate (cAMP) production and protein kinase A (PKA) activity, expression and phosphorylation of signalling molecules. Results Exogenous gAd increased melanin content, and the mRNA levels of microphthalmia-associated transcription factor (MITF) and its downstream genes TRP1, but not TRP2, were increased by gAd. However, cAMP production and PKA activity were not affected by gAd. Moreover, attempts to elucidate the underlying mechanism behind the gAd-mediated effect revealed that gAd could regulate melanogenesis by upregulating MITF through phosphorylation of the cAMP response element-binding protein (CREB). In addition, upregulation of MITF was mediated by activation of adenosine monophosphate-activated protein kinase (AMPK)-p38 mitogen-activated protein kinase (MAPK) signalling. Taken together, these findings indicate that promotion of melanogenesis by gAd occurs through increased expression of MITF, which is mediated by activation of the AMPK-p38 MAPK-CREB pathway. Conclusions These findings suggest that gAd contributes to epidermal homeostasis via its effect on melanocyte biology, and products of adipose tissue could affect epidermal biology.

Pulmonary fibroblasts are important structural lung cells that modulate tissue injury and immune response. It plays an important role in the inflammatory response by interacting between pulmonary fibroblasts and leukocytes. Cigarette smoke is known to trigger the development of diverse pulmonary and lung diseases, such as chronic obstructive pulmonary disease (COPD). α,β-unsaturated aldehydes such as acrolein (ACR) are present in cigarette smoke and are environment pollutants. ACR induces dysfunction of pulmonary and lung cells by forming DNA adducts and releasing inflammatory cytokines. MicroRNAs (miRNAs) are small interfering RNAs that negatively regulate mRNA gene expression and control the cellular response to toxic compounds. mRNAs also post-transcriptionally modulate gene expression. In this study, we identified whether the effects of ACR on the expression pattern of miRNAs in human pulmonary fibroblasts (HPFs). We carried out a pair-wise correlation analysis and examined 22 and 31 miRNAs with changed expression levels after treatment of HPFs with 10 μM and 25 μM ACR, respectively. Furthermore, we identified a significant anti-correlation in 102 and 17 mRNAs. Differentially expressed miRNAs and signaling pathways were further analyzed by gene ontology enrichment analysis. Our results showed that altered expression of miRNAs by ACR treatment was relevant in the dysfunction of respiratory cells and might contribute to the understanding of the mechanism of pulmonary diseases.

Cyclooxygenase-2 (COX-2), an inducible isoform protein, regulates diverse biological actions in vascular pathophysiology. COX-2 is induced in response to numerous stimuli, which results in prostaglandin (PG) production related to inflammation. Crotonaldehyde (CRA) is an extremely toxic α, β-unsaturated aldehyde and a major compound found in cigarette smoke. α, β-Unsaturated aldehyde in cigarette smoke is thought to mediate inflammation and vascular dysfunction. In this study, we evaluated the effect of CRA stimulation on COX-2 expression in human umbilical vein endothelial cells. CRA-stimulated COX-2 induction was accompanied by enhanced p38 phosphorylation and PGE2 generation. However, CRA-induced PGE2 production was reduced by pretreatment with an inhibitor of p38 MAPK. These results demonstrated that in human endothelial cells, CRA-induced COX-2-dependent PGE2 generation was mediated by p38 MAPK, and CRA may play a role in the development of inflammation.

Cardiovascular diseases are major causes of death worldwide, with pathologies including stroke, myocardial infarction, and vascular disease. Vascular grafts may be necessary to treat cardiovascular disease. The replacement of blood vessels has been very challenging in biomedical research. Dacron, ePTFE, and polyurethane are the most commonly used materials for artificial blood vessels. Bacterial cellulose (BC) synthesized by Gluconacetobacter xylinus exhibits unique properties including high purity, biocompatibility, resistance to degradation and low solubility. BC has been widely studied for applications in biomedical materials such as wound dressing and artificial skin. Recent studies have shown that BC is a promising material for preparing artificial blood vessels. This review provides a concise overview of current studies of BC and explores strategies for its clinical use.

MicroRNAs (miRNAs) play vital regulatory roles in various biological processes including cell differentiation, proliferation and apoptosis. Several studies have examined the expression profiles of miRNAs in response to different drugs and chemicals and suggested that expression patterns were associated with the effects of drugs and chemicals. Fisetin, a polyphenol found in a wide range of plants, has been reported to exhibit numerous biological activities, such as antioxidant, anti-inflammatory, antiangiogenic, and antiproliferative properties. In the present study, we evaluated whether fisetin regulates miRNA and mRNA expression. Using microarray analysis, we measured global miRNA and mRNA expression in human endothelial cells treated with 10 µM of fisetin for 24 h. Using several bioinformatic systems, we determined the signatures of potential biological processes and signaling pathways such as cell migration, apoptotic signaling pathway, cell motility, and cardiovascular system development. Our findings provide insight into the molecular mechanisms of action of fisetin and can be used to improve therapeutic intervention in vascular diseases.

Cigarette smoke contains approximately 5,000 chemical components, including reactive chemicals and free radicals that are associated with an increased risk of chronic obstructive pulmonary disease (COPD). Cigarette smoke is also known to promote inflammation as part of various inflammatory airway diseases. Crotonaldehyde (CRA) is a highly toxic α, β-unsaturated aldehyde that is a major component of cigarette smoke. Exposure to CRA has been previously shown to result in adverse effects on respiratory health. Heme oxygenase-1 (HO-1) is an inducible enzyme that is activated by various stress-inducing stimuli to perform a diverse range of physiological functions, including anti-oxidant, anti-apoptotic, and anti-inflammatory effects. The present study aimed to investigate the effect of CRA stimulation on HO-1 expression in human lung adenocarcinoma epithelial (A549) cells. Stimulation of cells with CRA was shown to result in upregulated HO-1 expression via the induction of nuclear factor erythroid 2-related factor 2 (Nrf2) nuclear translocation, and activation of the p38 mitogen-activated protein kinase (MAPK) pathway. Furthermore, zinc protoporphyrin (ZnPP; a specific HO-1 inhibitor) was used to inhibit HO-1 activity, and this was shown to cause a significant increase in the rate of apoptosis of CRA-exposed cells. Taken together, these results suggest that HO-1 exerts an anti-apoptotic effect in CRA-exposed human lung adenocarcinoma epithelial cells, and thus protects cells against CRA-induced oxidative stress.

Eriodictyol, a flavonoid commonly found in citrus fruits, shows neurotrophic, antioxidant, anti- inflammatory, and anti-cancer effects under diverse pathophysiological conditions such as vascular diseases. In this study, we evaluated whether eriodictyol modulates miRNA and mRNA expression. Using microarray analysis, we examined miRNA and mRNA expression in human endothelial cells treated with 10 μM of eriodictyol for 24 h. Using numerous bioinformatic systems, we evaluated the signatures of the potential biological processes and signaling pathways. Our results provide insight into the underlying molecular mechanisms of action of eriodictyol and suggest that eriodictyol can be used for therapeutic intervention in vascular disease.

Pulmonary fibroblasts are essential for the integrity of alveolar structures and to restore lung tissue after injury. They are also important for inflammatory responses through their ability to attract leukocytes. Cigarette smoke has many harmful components, and causes various pulmonary and lung diseases including chronic obstructive pulmonary disease (COPD), chronic bronchitis, and emphysema. Acrolein (ACR), one of the compounds in cigarette smoke, induces inflammatory cytokines and the generation of DNA adducts, resulting in dysfunction of respiratory cells such as pulmonary fibroblasts. In this study, we examined the expression of genes in ACR-treated human pulmonary fibroblasts by microarray profiling. We identified 2,378 and 312 genes that were differentially expressed within 6 h of treatment with 10 μM or 25 μM ACR, respectively. These genes were classified as being involved in many biological processes including apoptosis, immune responses, cell cycle, and signal transduction. Some genes, including HSPA1B, HMOX1, CASP3, PRDX3, and ANXA1, are related to COPD. These results support the hypothesis that ACR may increase cytotoxicity and tissue injury in respiratory cells and may attribute to the development of pulmonary disease.

Scaffold plays a critical role in stem cell differentiation and tissue regeneration. Composite scaffolds composed of bacterial cellulose (BC) and collagen (Col) in different ratios (1:1, 3:1, 5:1) were fabricated in this study. The composite scaffolds exhibit a well-organized interconnected porous structure, significantly better physical stability than Col scaffold, and more water uptake up to 400%. They were also favorable with cell attachment and growth. After osteogenic induction of umbilical cord blood derived mesenchymal stem cells (UCB-MSCs) for 3 weeks, we found more up-regulated osteogenic markers (collagen type 1, osteocalcin, bone sialoprotein) and significantly elevated proteins and calcium deposition, particularly with BC/Col (5:1) scaffold. When PKH-26 pre-labelled MSC-loaded scaffolds were subcutaneously transplanted in a mouse model, they showed many PKH-26-labelled cells and positive signals of α-smooth muscle actin, for neovascularization in the BC/Col (5:1). The current work demonstrates that our BC/Col composites may be promising as a bone tissue-engineered scaffold.

Crotonaldehyde (CRA), a reactive α,β-unsaturated aldehyde, is produced by natural and synthetic processes such as incomplete combustion of many compounds and can be found in cigarette smoke. CRA induces the formation of DNA adducts and suppresses the expression of glutathione, resulting in endothelial dysfunction, inflammation and vascular diseases. MicroRNAs (miRNAs) are small, non-coding RNAs that negatively modulate gene expression by targeting mRNAs for translational repression or degradation. They are also important factors involved in cell growth, apoptosis, and proliferation in cardiovascular disease. In this study, to examine the effect of CRA on the expression profiles of miRNAs in human umbilical vein endothelial cells (HUVECs), we performed pair-wise correlation analyses and identified 162 miRNAs with altered expression upon treatment of HUVECs with 10 μM CRA. In addition, we discovered a significant anti-correlation between 55 miRNAs and 11 mRNA. Differentially expressed miRNAs were further validated by Gene Ontology (GO) enrichment analysis. Our results suggest that modified expression of miRNAs caused by CRA treatment is related to endothelial dysfunction and might contribute to further understanding the molecular mechanisms of vascular diseases.

Melatonin is produced from tryptophan and is secreted by the pineal gland in periods of darkness; it has cytoprotective effects in endothelial cells. It has been reported that melatonin mediates the regulation of gene expression at the post-transcriptional level through its effect on miRNA expression. miRNAs are small non-coding RNAs that silence gene expression, regulate most cellular processes involved in development, and maintain cell functions. In this study, we confirmed the expression of 136 and 154 miRNAs when endothelial cells were treated with melatonin concentrations of 100 μM and 1 mM, respectively. We investigated anti-correlations in miRNA-target-mRNA pairs, and we identified a total of 13 and 44 mRNAs showing an expression difference of >1.5 fold, following exposure to melatonin concentrations of 100 μM and 1 mM, respectively. These profiling data revealed a variety of biological effects, including cell death, apoptosis and proliferation, via a Gene Ontology analysis of biological processes. The results of this study suggest that melatonin may regulate miRNA and gene expression and may have a cytoprotective effect on endothelial cells.

The cytotoxic drugs used in chemotherapy are often accompanied by nausea and vomiting. Despite the use of antiemetic drugs, chemotherapy-induced nausea and vomiting (CINV) remain significant side effects for cancer patients and are associated with serotonin type 3 receptor (5-HT3R) activation in the brainstem. Farnesol and nerolidol are sesquiterpene alcohols found in essential oils of plants such as roses, citronella, and lemon grass and are used as antiemetic parapheromones. Medicinal plants often are effective in treating gastrointestinal disorders, including CINV, although the mechanism of action remains unclear. In the current work, the antiemetic efficacy of the naturally occurring racemic mixture of farnesol (m-farnesol) and nerolidol (m-nerolidol) against cisplatin CINV was tested using the pica behavior (consumption of nonnutritive substances) of rats. Animals treated with m-farnesol or m-nerolidol consumed a smaller amount of kaolin than of saline-treated control animals. This result is consistent with the antiemetic efficacy of farnesol and nerolidol. Compared with controls, m-farnesol- but not m-nerolidol-treated animals consumed more food and lost less body weight. Thus, farnesol effectively reduced appetite suppression and weight loss induced by cisplatin. In separate experiments, isomers of farnesol and nerolidol were tested on 5-HT-induced responses of acutely isolated nodose neurons using patch-clamp methods. All the tested constituents inhibited 5-HT3R-mediated current in a noncompetitive manner. Thus, both farnesol and nerolidol may exert antiemetic efficacy by inhibiting 5-HT signaling in cranial visceral afferents, resulting in interruption of emetogenic signaling; however, nerolidol failed to suppress cisplatin-induced anorexia and weight loss, suggesting that additional mechanisms may contribute.

Background: Korean Red Ginseng (KRG) is a herbal medicine used in Asian countries and is very popular for its beneficial biological properties. Diabetes mellitus (DM) and its complications are rapidly becoming a global public health concern. The literature on transcriptional changes induced by KRG in rat models of diabetic retinopathy is limited. Considering these facts, we designed this study to determine whether retinopathy-associated genes are altered in retinas of rats with DM and whether the induced changes are reversed by KRG. Methods: Male Sprague–Dawley rats were intravenously injected with streptozotocin (50 mg/kg body weight) to induce DM, following which, KRG powder (200 mg/kg body weight) was orally administered to the KRG-treated DM rat group for 10 wks. The rats were then sacrificed, and their retinas were harvested for total RNA extraction. Microarray gene expression profiling was performed on the extracted RNA samples. Results: From among > 31,000 genes investigated, the expression of 268 genes was observed to be upregulated and that of 58 genes was downregulated, with twofold altered expression levels in the DM group compared with those in the control group. Moreover, 39 genes were upregulated more than twofold and 84 genes were downregulated in the KRG-treated group compared to the DM group. The expression of the genes was significantly reversed by KRG treatment; some of these genes were analyzed further to verify the results of the microarray experiments. Conclusion: Taken together, our data suggest that reversed changes in the gene expression may mediate alleviating activities of KRG in rats with diabetic retinopathy.

Cigarette smoking is one of the main sources of toxic chemical exposure to humans and is the greatest cause of progression of vascular disease. Crotonaldehyde is one of the major constituents of cigarette smoke and a product of endogenous lipid peroxidation. Thioredoxin reductase (TrxR) is a key element of the intact thioredoxin (Trx) system, which is predominant in modulating the intracellular redox homeostasis. In this study, we showed the effects of crotonaldehyde on the induction of TrxR1 expression in human endothelial cells. Crotonaldehyde exposure caused notably increased phosphorylation of p38 mitogen-activated protein kinase (MAPK). Introduction of siRNA against nuclear factor erythroid 2-related factor 2 (Nrf2) downregulated TrxR1 expression. Furthermore, treatment with auranofin, a TrxR inhibitor, significantly increased the death rate of crotonaldehyde-exposed cells. In summary, these results suggest a role for TrxR1 upregulation through p38 MAPK-Nrf2 activation in the adaptive response of human endothelial cells to crotonaldehyde.

Atopic dermatitis (AD) is a common multifactorial chronic skin disease that has a multiple and complex pathogenesis. AD is gradually increasing in prevalence globally. In NC/Nga mice, repetitive applications of 2, 4-dinitrofluorobenzene (DNFB) evoke AD-like clinical symptoms similar to human AD. Aspartame (N-L-α-aspartyl-L-phenylalanine 1-methyl ester) is a methyl ester of a dipeptide, which is used as an artificial non-nutritive sweetener. Aspartame has analgesic and anti-inflammatory functions that are similar to the function of nonsteroidal anti-inflammatory drugs such as aspirin. We investigated whether aspartame can relieve AD-like clinical symptoms induced by DNFB treatment in NC/Nga mice. Sucrose did not relieve AD-like symptoms, whereas aspartame at doses of 0.5 μg kg(-1) and 0.5 mg kg(-1) inhibited ear swelling and relieved AD-like clinical symptoms. Aspartame inhibited infiltration of inflammatory cells including eosinophils, mast cells, and CD4(+) T cells, and suppressed the expression of cytokines including IL-4 and IFN-γ, and total serum IgE levels. Aspartame may have therapeutic value in the treatment of AD.

MicroRNAs (miRNAs) are associated with a diverse range of biological processes, human diseases, and metabolic disorders. Myricetin, which is the most abundant polyphenol class in the human diet, has antioxidative, antiapoptotic, and anti-inflammatory properties. The present study aimed to evaluate whether myricetin modulates miRNA expression. Using microarray analysis, we investigated miRNA expression in human endothelial cells treated with 25 µM and 100 µM of myricetin for 24 hours, and found that 101 and 191 miRNAs, respectively, were differential expressed by at least 1.5-fold. Based on several bioinformatic systems, we also identified signatures of the potential biological processes and signaling pathways that are influenced by dysregulated miRNAs. Therefore, integrating specific patterns of miRNA and mRNA levels may suggest a new mechanism of action of myricetin.

Ethnopharmacological relevance Gentiana scabra root extract (GS) is frequently prescribed as an internal remedy in traditional Korean medicine for treatment of diabetes mellitus. GS contains bitter iridoid glycosides including loganic acid, gentiopicrin, trifloroside, and rindoside. We previously reported that the intestinal bitter taste sensation stimulates GLP-1 secretion, and thereupon hypothesized that the blood glucose regulatory effect of GS is due to its GLP-1 secreting effect in enteroendocrine L cells. Materials and method We studied GLP-1 secreting effect of GS treatment and its cellular downstream mechanism in human enteroendocrine NCI-H716 cells using the G protein-coupled receptor (GPCR) pathway inhibitors. Intracellular calcium assay also demonstrated the signal transduction pathway stimulated by the GS treatment. Using db/db mice, we performed oral glucose tolerance test (OGTT) to examine the blood glucose lowering effect of GS administration. We also collected the mouse plasma during the OGTT to measure the GLP-1 and insulin levels. Result We demonstrated dose-dependent GLP-1 secreting effect of GS on the NCI-H716 cells. The GLP-1 secreting effect of GS is mediated by the G protein βγ-subunit and inositol triphosphate. Using db/db mice, we found that the effect of GS on lowering blood glucose is due to its GLP-1 secretion, and consequential insulinotropic effect. The chemical fingerprint of GS was obtained through a direct analysis in realtime mass spectrometry (DART-MS) and high-performance liquid chromatography (HPLC)/MS. Through the GLP-1 secretion study, we found that loganic acid, an iridoid glycoside, contributes to the GLP-1 secreting effect of GS. Conclusion The findings of this study highlight the potential of exploiting the antidiabetic effect of GS on type 2 diabetes mellitus patients.

Cigarette smoke represents one of the most significant risk factors for the progression of vascular disease. Exposure to cigarette smoke induces oxidative stress in the vascular system and results in vascular dysfunction. Crotonaldehyde, a highly reactive α, β-unsaturated aldehyde, is a foremost compound of cigarette smoke and a product of endogenous lipid peroxidation. Heme oxygenase-1 (HO-1) expression plays an essential role in cellular defense against environmental insults and represents an adaptive response. Here, we showed the effects of crotonaldehyde on the induction of HO-1 expression in RAW 264.7 macrophages. Crotonaldehyde treatment resulted in significantly increased phosphorylation of p38 mitogen-activated protein kinase (MAPK) and nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2). Furthermore, treatment with zinc protoporphyrin (ZnPP; the specific HO-1 inhibitor) markedly augmented the death of crotonaldehyde-treated macrophages. In summary, these results highlight the role of the HO-1 upregulation through p38 MAPK-Nrf2 activation in the adaptive response to crotonaldehyde in macrophages.

MicroRNAs (miRNAs) have been implicated in the regulation of physiological and pathophysiological processes such as vascular disease. Analysis of miRNA expression profiles in vascular disease provides new insights into the possible mechanisms underlying and therapeutic targets of disease. Acrolein is a highly reactive component of cigarette smoke, which has been implicated in the development of vascular disease. In this study, we investigated whether miRNAs play a role in the modulation of various gene expression responses to acrolein in human endothelial cells. We analyzed whole-genome miRNA and mRNA expression and found that acrolein induced differential expression of 151 miRNAs in endothelial cells. We further classified a number of target genes of the differentially expressed miRNAs. Among the differentially expressed miRNAs, miR-342-3p and 422a were upregulated, and the expression levels demonstrated a high inverse correlation with their targets, the vascular disease- related genes CLDN1 and PON1. Thus, integrating specific patterns of miRNA and mRNA levels may improve the diagnosis of vascular diseases induced by exposure to environmental pollutants such as acrolein.

Of the biogenic polyamines, spermidine is a natural constituent of living cells and organisms. Spermidine is associated with regulation of cell growth, proliferation and differentiation, and with the suppression of oxidation and inflammation. Atopic dermatitis (AD) is a chronic inflammatory skin disease that has a complex and multiple pathogenesis, which includes genetic abnormality, modified or abnormal immune response and the production of nitric oxide and reactive oxygen species. We investigated whether spermidine can relieve AD-like clinical manifestation induced by the continual application of 2,4-dinitrofluorobenzene (DNFB) in NC/Nga mice. Spermidine at concentrations of 1 or 10 mg/kg reduced increasing ear swelling and attenuated oedema, haemorrhage and hyperkeratosis in AD-like skin lesions. Repetitive application of DNFB induced inflammatory cell infiltration to skin lesions, whereas intraperitoneal injection of spermidine inhibited DNFB-evoked infiltration of eosinophils, mast cells and T lymphocytes. Furthermore, spermidine suppressed mast cell degranulation and production of interferon-gamma by activated CD4(+) T cells in AD-like skin lesions. Spermidine may be a potential therapeutic agent for treatment of AD.

Ultraviolet A (UVA) irradiation exerts negative effects on stemness and differentiation potential of stem cells. This study aimed to explore the effect of sinapic acid on UVA-irradiation-induced damages to stemness and differentiation potential of human-adipose-tissue-derived mesenchymal stem cells (hAMSCs) and its UVA-antagonist mechanisms. Sinapic acid attenuated UVA-induced reduction in the proliferative potential and stemness by upregulating OCT4, SOX2, and NANOG. In addition, sinapic acid significantly recovered UVA-induced reduction in expression level of hypoxia-inducible factor (HIF)-1α. The antagonist effect of sinapic acid against stemness damage was mediated by reduceing PGE2 production through inhibition of p38 MAPK and NF-κB. Moreover, sinapic acid attenuated UVA-induced reduction in differentiation potential by downregulating the expression of macrophage migration inhibitory factor (MIF) and Kruppel-like factor (KLF) 2 gene while activating AMP-activated protein kinase (AMPK). UVA-induced inhibition of adipogenic differentiation was mediated by reducing MIF production through suppression of NF-κB. Taken together, these findings suggest that sinapic acid may ameliorate UVA-irradiation-induced reduced stemness and differentiation potential of hAMSCs. Therefore, sinapic acid might have potential as an antagonist agent to attenuate damages caused by UVA.

The common environmental contaminant 3-methylcholanthrene (3MC) is found in cigarette smoke and is produced by incomplete combustion of fat, wood and coal. 3MC is a poly aromatic hydrocarbon that interacts with an aryl hydrocarbon receptor and causes inflammation and induces vascular dysfunction; 3MC forms the DNA adducts structure and regulates cell cycle, which increase oxidative stress and inflammation. MicroRNAs (miRNAs) are non-coding RNA molecules that negatively regulate gene expression; these RNAs also play a role in cellular and molecular responses to toxicants, and can post-transcriptionally regulate gene expression. In this study, we examined whether miRNAs affect the regulation of gene expression in 3MC-treated human umbilical vein endothelial cells (HUVECs). We carried out pair-wise correlation analysis and identified 131 and 116 miRNAs with altered expression upon treatment of HUVECs with 100 nM and 1 μM 3MC, respectively. Furthermore, we identified 188 and 85 mRNAs with altered expression upon treatment with 100 nM and 1 μM 3MC; we subsequently analyzed their anti-correlations. The Gene Ontology (GO) enrichment analysis on the altered expression of miRNA-related genes displayed significant enrichment for genes involved in certain biological processes. Specifically, our results suggest that changes in miRNA expression caused by 3MC treatment are associated with inflammation of endothelial cells and may play a role in cardiovascular disease.

Melatonin is secreted by the pineal gland and gastrointestinal tract and plays a role in circadian rhythm synchronization. Melatonin is also reported to have various physiological effects on endothelial cells. Endothelial cells are responsible for physiologic homeostasis and maintaining blood vessel permeability in vivo. In this study, using gene expression profiling, we confirmed about 151 up-regulation and 1,023 downregulation expression genes, with an expression level difference of > 1.5-fold, after exposure to 100 μM and 1 mM melatonin concentrations in endothelial cells. The genes were classified into 16 functional groups based on their functions, including cell adhesion, immune response, lipid metabolism, and response to stress. We analyzed the gene expression profiling data of melatonin-treated human umbilical vein endothelial cells (HUVECs), which indicated a broad spectrum of physiological functions, including anti-oxidative/anti-inflammatory effects and cell-to-cell junction maintenancerelated genes.

Atopic dermatitis (AD) is a multifactorial chronic skin disorder that is increasing in prevalence globally. In NC/Nga mice, repetitive epicutaneous applications of 2-4-dinitrofluorobenzene (DNFB) induces AD-like clinical symptoms. Bioflanonol fisetin (3,7,3',4'-tetrahydroxyflavone) is a dietary component found in plants, fruits and vegetables. Fisetin has various physiological effects that include anti-oxidation, anti-angiogenesis, anti-carcinogenesis and anti-inflammation. In this study, we investigated whether fisetin relieves AD-like clinical symptoms induced by repeated DNFB treatment in NC/Nga mice. Fisetin significantly inhibited infiltration of inflammatory cells including eosinophils, mast cells and CD4(+) T and CD8(+) T cells, and suppressed the expressions of cytokines and chemokines associated with dermal infiltrates in AD-like skin lesions. Total serum immunoglobulin E (IgE) levels and the ratio of phospho-NF-κB p65 to total NF-κB p65 were markedly reduced by fisetin. Fisetin also reduced the production of interferon-gamma and interleukin-4 by activated CD4(+) T cells in a dose-dependent manner, whereas the anti-inflammatory cytokine, interleukin-10 was increased. These results implicate fisetin as a potential therapeutic for AD.

Nausea and emesis are a major side effect and obstacle for chemotherapy in cancer patients. Employ of antiemetic drugs help to suppress chemotherapy-induced emesis in some patients but not all patients. Ginger, an herbal medicine, has been traditionally used to treat various kinds of diseases including gastrointestinal symptoms. Ginger is effective in alleviating nausea and emesis, particularly, for cytotoxic chemotherapy drug-induced emesis. Ginger-mediated antiemetic effect has been attributed to its pungent constituents-mediated inhibition of serotonin (5-HT) receptor activity but its cellular mechanism of action is still unclear. Emetogenic chemotherapy drugs increase 5-HT concentration and activate visceral vagal afferent nerve activity. Thus, 5-HT mediated vagal afferent activation is essential to provoke emesis during chemotherapy. In this experiment, water extract of ginger and its three major pungent constituent's effect on 5-HT-evoked responses were tested on acutely dispersed visceral afferent neurons with patch-clamp methods. The ginger extract has similar effects to antiemetic drug ondansetron by blocking 5-HT-evoked responses. Pungent constituents of the ginger, [6]-shogaol, [6]-gingerol, and zingerone inhibited 5-HT responses in a dose dependent manner. The order of inhibitory potency for these compounds were [6]-shogaol>[6]-gingerol>zingerone. Unlike well-known competitive 5-HT3 receptor antagonist ondansetron, all tested ginger constituents acted as non-competitive antagonist. Our results imply that ginger and its pungent constituents exert antiemetic effects by blocking 5-HT-induced emetic signal transmission in vagal afferent neurons.

Cigarette smoke has many toxic components that can lead to the disruption of various cellular processes. Increase in oxidative stress due to cigarette smoke causes production of reactive oxygen species, which induce vascular disease. 3-methylcholanthrene (3MC) is an environmental contaminant derived from the incomplete combustion of cigarette smoke and is one of the major polycyclic aromatic hydrocarbons. 3MC activates the aryl hydrocarbon receptor and contributes to carcinogenesis and vascular disease. The present study investigated gene expression in 3MC-treated human umbilical vein endothelial cells by microarray gene expression profiling. We found 1,279 and 717 genes with altered gene expression within 24 h of 100 nM and 1 μM 3MC treatment, respectively; these genes were distributed into distinct functional groups. Some of the genes including ICAM1, PTGS2, APOL3, and GPX-1, were related to cardiovascular disease. Our results support the hypothesis that 3MC-stimulated gene expression in endothelial cells may play a role in the development of cardiovascular disease.

Myricetin is a dietary flavonoid that is found in many fruits, vegetables, teas and medicinal herbs. It has been demonstrated to have antioxidative, antiapoptotic, and anti-inflammatory properties, but the precise mechanism underlying the effects of myricetin at the transcriptional level has not yet been determined. The present study aimed to determine the target genes of myricetin in human umbilical vein endothelial cells (HUVECs) by microarray gene expression profiling. Among the more than 40,000 genes investigated, 121 and 953 genes were upregulated by at least 1.5-fold, while 584 and 1426 genes were downregulated by at least 1.5-fold in 24 h for 25 μM and 100 μM myricetin treatment. Gene ontology (GO) analysis showed that the top biological processes disturbed in HUVECs were regulation of apoptosis and cellular metabolic processes. Furthermore, vascular diseaserelated genes including HIRA, HDAC9, HIF1A, and RTN3 were also altered by myricetin. These results demonstrate myricetin-induced genome-wide alterations in the expression profile of HUVECs and suggest that myricetin exerts a vasculoprotective effect through transcriptional changes in the human endothelium. In conclusion, our data support the view that myricetinstimulated changes in gene expression contribute to protective effects on vascular disease.

Prosthetic biomaterials are required to be non-toxic, non-thrombogenic, and non-immunogenic. Bacterial cellulose (BC) synthesized by Gluconacetobacter xylinus has recently been studied as a biocompatible material due to its unique features such as high purity, crystallinity, biodegradability, and tensile strength as compared to plant cellulose. Although BC has high potential to be used as biomaterial, its toxicity and immunoreactivity have not been properly studied yet. In this report, we investigated the immunoreactivity of BC in vitro in human umbilical vein endothelial cells (HUVECs) and in vivo using BALB/c mice. We report that BC does not induce apoptosis and necrosis in HUVECs and does not stimulate immune response in both HUVECs and BALB/c mice models. These results suggest that BC may be widely used as a biocompatible biomaterial for tissue engineering and biosensors.

Methylglyoxal (MG) is a highly toxic compound that contributes to the formation of advanced glycation end products (AGEs). MG is detected at high concentration in plasma of diabetic patients and is involved in the pathogenesis of several diabetic macro- and microvascular complications, such as atherosclerosis, retinopathy, nephropathy and hypertension. MicroRNAs (miRNAs) play a critical role in the negative regulation of genes in many biologic processes. They are also known to post-transcriptionally regulate gene expression involved in cellular responses to toxicants. In this study, we investigated whether miRNAs play a role in the regulation of gene expressions in human umbilical vein endothelial cells (HUVECs) after the treatment with MG. We performed pair-wise correlation analysis, acquired altered expression levels of 274 miRNAs and 686 mRNA, and observed their anti-correlations. Genes associated with diabetic vascular disease were sorted from miRNA-correlated genes, and Gene Ontology (GO) enrichment analysis on the differential expression of selected genes was carried out. Our results revealed the relationship between miRNAs and mRNA in MG-exposed endothelial cells using the expression profiling.

Acrolein (ACR) is a highly reactive α, β-unsaturated carbonyl compound that is related to several diabetic vascular complications. ACR, a ubiquitous aldehyde pollutant, is a product of incomplete combustion reactions and a major component of cigarette smoke. ACR is also endogenously generated during lipid peroxidation. Glycosylphosphatidylinositol (GPI)-anchored proteins is a membrane bound protein, which have a variety of cellular functions. It is reported that some of them showed the reduced level under diabetic conditions. In this study, we explored whether ACR may affect GPI anchor related gene expression in human umbilical vein endothelial cells (HUVECs) by modulation of miRNA expression that play a significant role in the post-transcriptional regulation of genes. We performed global gene expression analysis, acquired altered expression levels of 27 miRNAs and 21 mRNAs that are relevant to GPI anchored proteins, and observed their anti-correlations. Gene Ontology (GO) terms that are closely associated with GPI anchor biosynthesis were highest when analysis was based on biological processes characterized by enriched GO categories. Our results revealed that ACR may control miRNAs to regulate genes involving in GPI anchor biosynthesis in human endothelial cells.

Spermidine (SPD) is a ubiquitous polycation that is commonly distributed in living organisms. Intracellular levels of SPD are tightly regulated, and SPD controls cell proliferation and death. However, SPD undergoes oxidation in the presence of serum, producing aldehydes, hydrogen peroxide, and ammonia, which exert cytotoxic effect on cells. Hemeoxygenase-1 (HO-1) is thought to have a protective effect against oxidative stress. Upregulation of HO-1 in endothelial cells is considered to be beneficial in the cardiovascular disease. In the present study, we demonstrate that the ubiquitous polyamine, SPD, induces HO-1 in human umbilical vein endothelial cells (HUVECs). SPD-induced HO-1 expression was examined by Western blot and reverse transcription-polymerase chain reaction (RT-PCR). Involvement of reactive oxygen species, serum amine oxidase, PI3K/Akt signaling pathway, and transcription factor Nrf2 in the induction of HO-1 by SPD was also investigated. Furthermore, small interfering RNA knockdown of Nrf2 or HO-1 and treatment with the specific HO-1 inhibitor ZnPP exhibited a noteworthy increase of death of SPD-stimulated HUVECs. In conclusion, these results suggest that SPD induces PI3K/Akt-Nrf2-mediated HO-1 expression in human endothelial cells, which may have a role in cytoprotection of the cells against oxidative stress-induced death.

Tissue regeneration using adult stem cells (ASCs) has significant potential as a novel treatment for many degenerative diseases. Previous studies have established that age negatively affects the proliferation status and differentiation potential of ASCs, suggesting a possible limitation in their potential therapeutic use. Therefore, we hypothesized that apple extract might exert beneficial effects on ASCs. The specific objectives were to investigate the proliferative effect of apple ethanol extract on human adipose tissue-derived mesenchymal stem cells (ADSCs) and human cord blood-derived mesenchymal stem cells (CB-MSCs), and identify the possible molecular mechanisms. Apple extract promoted proliferation of ADSCs and CB-MSCs as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Click-iT 5-ethynyl-2'-deoxyuridine flow cytometry assays. In addition, phosphorylation of p44/42 MAPK (ERK), mammalian target of rapamycin (mTOR), p70 S6 kinase (p70S6K), S6 ribosomal protein (S6RP), eukaryotic initiation factor (eIF) 4B and eIF4E was induced stepwise in ADSCs. Furthermore, apple extract significantly induced the production of vascular endothelial growth factor and interleukin-6 in both ADSCs and CB-MSCs. Similarly, apple extract-induced phosphorylation of the mTOR/p70S6K/S6RP/eIF4B/eIF4E pathway was blocked by pretreatment with PD98059, a specific ERK inhibitor. These results indicate that apple extract-induced proliferation of ADSCs under serum-free conditions is mediated by ERK-dependent cytokine production. Moreover, the beneficial effect of apple extract on proliferation of ASCs may overcome the limitation in therapeutic use of stem cells in tissue regeneration and maintenance of stem cell homeostasis.

Atopic dermatitis (AD) is a common inflammatory skin disease for which few effective treatments are available. Resolvin E1 (RvE1; 5S,12R,18R-trihydroxy-6Z,8E,10E,14Z,16E-eicosapentaenoic acid) is an endogenous lipid mediator derived from omega-3 fatty eicosapentaenoic acid, which is a potent inhibitor of inflammation. AD-like skin lesion was induced by repetitive skin contact with DNFB in NC/Nga mice and the effects of RvE1 were evaluated on the basis of histopathological findings of skin, ear swelling and cytokine production of CD4(+) T cells. Intraperitoneal injection of RvE1 for one week after DNFB challenge significantly lowered ear swelling and improved back skin lesions. In addition, RvE1 significantly suppressed production of interferon-gamma (IFN-γ) and interleukin-4 (IL-4) by activated CD4(+) T cells and serum IgE level. Furthermore, RvE1 reduced DNFB-induced infiltration of eosinophils, mast cells, CD4(+) T cells, and CD8(+) T cells in skin lesions. Therefore, RvE1 may suppress the development of AD-like skin lesions in DNFB-treated NC/Nga mice by reducing IL-4 and IFN-γ of activated CD4(+) T cells and serum IgE levels and infiltration of immune cells to skin lesion.