RESUMO
Fabry disease is an X-linked lysosomal storage disorder that is caused by a deficiency of α-galactosidase A. The disease ultimately manifests as multiple organ dysfunctions owing to excessive accumulation of globotriaosylceramide (Gb3). Among the several complications of Fabry disease, ascending thoracic aortic aneurysm is relatively common, which is classically associated with connective tissue disorders characterized by abnormal defects or deficiencies in structural proteins such as collagen and elastin. Although an elevated Gb3 level is regarded as a prerequisite for the manifestations of Fabry disease, only this excess accumulation cannot explain the pathophysiology of these complications. Recently, an increased plasma level of lyso-Gb3 was suggested as a new biomarker in Fabry disease. Therefore, the aim of this study was to assess the effects of lyso-Gb3 on the pathogenesis of thoracic ascending aortic aneurysms in Fabry disease, with a particular focus on the responses related to aortic remodeling by fibroblasts. We found that lyso-Gb3 inhibited the growth of fibroblasts, as well as their differentiation into myofibroblasts, and collagen expression. Moreover, all of these compromised responses could be attributed to the effects of lyso-Gb3 on downregulation of KCa3.1 channel expression, and these impairments could be rescued when activating the KCa3.1 channel or increasing intracellular Ca(2+) concentration. This study provides new evidence that lyso-Gb3 inhibits the differentiation into myofibroblasts and collagen synthesis of fibroblasts owing to decreased Ca(2+) levels by KCa3.1 channel dysfunction. These findings suggest that the KCa3.1 channel can serve as a new target to attenuate and prevent development of ascending thoracic aortic aneurysm in Fabry disease.
Assuntos
Cálcio/metabolismo , Colágeno/biossíntese , Fibroblastos/citologia , Fibroblastos/fisiologia , Glicolipídeos/metabolismo , Canais de Potássio Ativados por Cálcio de Condutância Intermediária/metabolismo , Esfingolipídeos/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/fisiologia , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Glicolipídeos/administração & dosagem , Camundongos , Células NIH 3T3 , Esfingolipídeos/administração & dosagemRESUMO
Neutropenia is a principal complication of cancer treatment. We investigated the supportive effect of adipose tissue-derived mesenchymal stem cells (AD-MSCs) on the viability and function of neutrophils. Neutrophils were derived from HL-60 cells by dimethylformamide stimulation and cultured with or without AD-MSCs under serum-starved conditions to evaluate neutrophil survival, proliferation, and function. Serum starvation resulted in the apoptosis of neutrophils and decreased cell survival. The co-culture of neutrophils and AD-MSCs resulted in cell survival and inhibited neutrophil apoptosis under serum-starved conditions. The survival rate of neutrophils was prolonged up to 72 h, and the expression levels of interferon (IFN)-α, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor, and transforming growth factor (TGF)-ß in AD-MSCs were increased after co-culture with neutrophils. AD-MSCs promoted the viability of neutrophils by inhibiting apoptosis as well as enhancing respiratory burst, which could potentially be mediated by the increased expression of IFN-α, G-CSF, and TGF-ß. Thus, we conclude that the use of AD-MSCs may be a promising cell-based therapy for increasing immunity by accelerating neutrophil function.
Assuntos
Técnicas de Cocultura , Células-Tronco Mesenquimais/fisiologia , Neutrófilos/imunologia , Tecido Adiposo/citologia , Diferenciação Celular , Sobrevivência Celular/genética , Expressão Gênica , Fator Estimulador de Colônias de Granulócitos/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Células HL-60 , Humanos , Interferon-alfa/imunologia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Neutropenia/terapia , Neutrófilos/citologia , Neutrófilos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fatores de Crescimento Transformadores/imunologiaRESUMO
Vivax malaria re-emerged in the Republic of Korea (ROK) in 1993. The annual incidence of this disease, which had increased rapidly through 2000 with geographic expansion, started to decrease in 2001, reaching 864 cases in 2004; however, the trends changed in 2005 when 1,304 cases were reported. Among 2,168 cases of vivax malaria reported from 2004 through 2005, 389 cases (17.9%) were ROK military personnel, 565 cases (26.1%) were veterans who had been discharged from the military within 2 years of report of infection, and 1,214 cases (56.0%) were civilians. Local transmission might have taken place during this period in the southern side of the Demilitarized Zone. Regional increase of vivax malaria in North Korea, increased local transmissions in ROK, and active transmission by vector mosquitoes during the transmission season might be important factors responsible for the re-increase of vivax malaria in ROK during 2005.
Assuntos
Malária Vivax/epidemiologia , Militares , Animais , Anopheles/fisiologia , Humanos , Incidência , Insetos Vetores/fisiologia , Coreia (Geográfico)/epidemiologia , Plasmodium vivax , Densidade Demográfica , Vigilância da População , Fatores de TempoRESUMO
VvpM, one of the extracellular metalloproteases produced by Vibrio vulnificus, induces apoptotic cell death via a pathway consisting of ERK activation, cytochrome c release, and activation of caspases-9 and -3. VvpM-treated cells also showed necrotic cell death as stained by propidium iodide (PI). The percentage of PI-stained cells was decreased by pretreatment with Necrostatin-1, indicating that VvpM-mediated cell death occurs through necroptosis. The appearance of autophagic vesicles and lipidated form of light-chain-3B in rVvpM-treated cells suggests an involvement of autophagy in this process. Therefore, the multifarious action of VvpM might be one of the factors responsible for V. vulnificus pathogenesis.
Assuntos
Apoptose , Autofagia , Morte Celular , Metaloproteases/metabolismo , Vibrio vulnificus/enzimologia , Vibrio vulnificus/patogenicidade , Caspase 3/metabolismo , Caspases/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular , Citocromos c/metabolismo , Humanos , Imidazóis/farmacologia , Indóis/farmacologia , Necrose , Vibrio vulnificus/metabolismoRESUMO
Zinc is considered to be involved in maintaining healthy vascular condition. Atherosclerotic calcification of vascular smooth muscle cells (VSMCs) occurs via the mechanism of cell death; therefore, cell viability is a critical factor for preventing VSMC calcification. In this study, we tested whether zinc affected VSMC viability under both normal physiological non-calcifying (0 mM P) and atherosclerotic calcifying conditions (3 and 5 mM P), since VSMC physiological characters change during the VSMC calcification process. The study results showed that an optimal zinc level (15 µM) restored the decreased VSMC viability which was induced under low zinc levels (0 and 1 µM) and calcifying conditions (3 and 5 mM P) at 9 and 15 days culture. This zinc-protecting effect for VSMC viability is more prominent under atherosclerotic calcifying condition (3 and 5 mM P) than normal condition (0 mM P). Also, the increased VSMC viability was consistent with the decreased Ca and P accumulation in VSMC cell layers. The results suggested that zinc could be an effective biomineral for preventing VSMC calcification under atherosclerotic calcifying conditions.
RESUMO
A pathogenic bacterium, Vibrio vulnificus produces various extracellular proteases including the elastolytic metalloprotease, VvpE. In silico analysis of its genome revealed a VvpE-homologous protease, VvpM whose proteolytic activity was abolished by specific inhibitors against metalloproteases. To investigate whether this newly identified protease has pathogenic role in host interaction in addition to proteolytic role, human cell lines were incubated with recombinant VvpM (rVvpM). rVvpM-challenged cells showed typical morphological changes found in cells under apoptosis. Apoptotic cell death was further evidenced by estimating the Annexin V-stained cells, whose proportions were dependent upon the concentrations of rVvpM treated to human cells. To elucidate the signaling pathway for VvpM-induced apoptosis, three MAPKs were tested if their activation were mediated by rVvpM. ERK1/2 was phosphorylated by treatment of rVvpM and rVvpM-induced cell death was blocked by a specific inhibitor against ERK1/2. In rVvpM-treated cells, the cytosolic levels of cytochrome c were increased in a VvpM concentration-dependent manner, while the levels of cytochrome c in mitochondria were decreased. Cell deaths were accompanied by apparent cleavages of procaspases-9 and -3 to the active caspases-9 and -3, respectively. Therefore, this study demonstrates that an extracellular metalloprotease of V. vulnificus, VvpM induces apoptosis of human cells via a pathway consisting of ERK activation, cytochrome c release, and then activation of caspases-9 and -3.
Assuntos
Apoptose , Proteínas de Bactérias/metabolismo , Metaloproteases/metabolismo , Vibrio vulnificus/enzimologia , Vibrio vulnificus/patogenicidade , Sequência de Aminoácidos , Anexina A5/análise , Apoptose/fisiologia , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Caspase 3/metabolismo , Caspase 9/metabolismo , Simulação por Computador , Citocromos c/metabolismo , Citosol/metabolismo , Ativação Enzimática , Células HCT116 , Células HT29 , Humanos , Metaloproteases/química , Metaloproteases/genética , Metaloproteases/isolamento & purificação , Microscopia Eletrônica de Transmissão , Mitocôndrias/metabolismo , Dados de Sequência Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Transdução de SinaisRESUMO
Yam (Dioscorea batatas) is widely consumed as functional food for health promotion mainly in East Asia countries. We assessed whether yam root (tuber) or bark (peel) extracts stimulated the activity of osteoblasts for osteogenesis. MC3T3-E1 cells (mouse osteoblasts) were treated with yam root extracts (water or methanol) (study I) or bark extracts (water or hexane) (study II) within 0~10 µg/mL during the periods of osteoblast proliferation (5~10 day), matrix maturation (11~15 day) and mineralization (16~20 day) as appropriate. In study I, both yam root water and methanol extracts increased cell proliferation as concentration-dependent manner. Cellular collagen synthesis and alkaline phosphatase (ALP) activity, both the indicators of bone matrix protein and inorganic phosphate production for calcification respectively, were also increased by yam root water and methanol extract. Osteoblast calcification as cell matrix Ca and P accumulation was also increased by the addition of yam root extracts. In study II, yam bark extracts (water and hexane) increased osteoblast proliferation and differentiation, as collagen synthesis and ALP activity and osteoblast matrix Ca and P deposition. The study results suggested that both yam root and bark extracts stimulate osteogenic function in osteoblasts by stimulating bone matrix maturation by increasing collagen synthesis, ALP activity, and matrix mineralization.
RESUMO
The calcification of vascular smooth muscle cells (VSMCs) is considered one of the major contributors for vascular disease. Phosphate is known as the inducer for VSMC calcification. In this study, we assessed whether phosphate affected cell viability and fetuin-A, a calcification inhibitor protein, both which are related to VSMC calcification. Also, VSMC viability by zinc level was assessed. The results showed that phosphate increased Ca and P deposition in VSMCs (A7r5 cell line, rat aorta origin). This phosphate-induced Ca and P deposition was consistent with the decreased A7r5 cell viability (P<0.05), which implies phosphate-induced calcification in A7r5 cells might be due to the decreased VSMC cell viability. As phosphate increased, the protein expression of fetuin-A protein was up-regulated. A7r5 cell viability decreased as the addition of cellular zinc level was decreased (P<0.05). The results suggested that zinc deficiency causes the decreased cell viability and it would be the future study to clarify how zinc does act for VSMC cell viability. The results suggest that the decreased VSMC viability by high P or low Zn in VSMCs may be the risk factor for vascular disease.
RESUMO
Yam (Dioscorea) has long been used as foods and folk medicine with the approved positive effects for health promotion. Although consumption of yam products is increasing for health promotion, reports for the metal contamination in commercial yam powder products to protect the consumers are lacking. In this study, we aimed to assess whether the commercial yam powder products were heavy metal contaminated or not using the yam products from six commercial products from various places in South Korea. The contents of heavy metals (Cd, Cr, As, Pb, Ni, and Sn) in yam powder products were measured and compared to national and international food standard levels. Also, the metal contamination was monitored during the food manufacturing steps. The study results showed that the contents of heavy metals (Cd, Cr, As, and Pb) in yam powder products are similar to those in national 'roots and tubers' as well as in various crops. In comparison to three international standard levels (EU, Codex and Korea), Cd content in yam powder products was lower but Pb content was 5 times higher. Also, Pb, Ni, and Sn may have the potential to be contaminated during food manufacturing steps. In conclusion, the level of heavy metals (Cd, Cr, As, Ni, and Sn) except Pb is considered relatively safe on comparison to national and international food standard levels.
RESUMO
Yam has been recognized having the beneficial effects for the prevention of various diseases, such as cancer, immunity, infection and obesity etc. There is increasing consideration to supplement the antioxidant nutrients to make up the lack of the antioxidant nutrient intakes. No study has been reported for the analysis of antioxidant mineral contents and comparison to dietary recommended intake for the sense of health promotion. In our study, we analyzed the contents of antioxidant trace elements (Zn, Mn, Fe, Cu and Se) and Cr contents in cultivated Korean yam powders for evaluation of nutrient intake aspects. We collected the commercial yam powders from six different cultivated areas in the South Korea and measured antioxidant minerals (Zn, Mn, Fe, Cu and Se) and Cr contents using trace element-free plasma spectrometer (ICP) or atomic absorption spectrometer (AAS) after dry-ashing and then wet-acid digestion. The accuracy of mineral analysis method was confirmed by the mineral analysis of standard reference material. Each analyzed element contents in yam were compared to dietary reference intakes of Koreans (KDRIs). The average levels of trace elements (Zn, Mn, Fe, Cu, Se and Cr) in yam powders were 18.3, 11.9, 36.0, 3.7, 1.9 and 1.27 µg/g yam powder, respectively. The intakes of Zn, Fe, Cu and Se of which KDRIs is determined, are accounted as being up to 23.8%, 55.6%, 32.5% and 236% recommended intake (RI) of KDRIs, if daily yam supplementation (50 g) of commercial instruction would be considered. The intake of Mn is about 25% adequate intake (AI) of KDRIs with the daily supplementation of yam powder. Most of mineral intakes from daily yam supplementation were with the range of non-detectable to <10% upper limit (UL) level, which is very much safe. The study results show that daily supplementation of Korean yam power is beneficial to provide the supplemental nutrient intake and also is safe, if the suggested dosage would be considered.
RESUMO
Diosgenin, a steroid saponin extracted from the root of wild yam (Dioscorea villossa) is claimed to have osteogenic property. However, detailed studies providing evidence to this claim have not been fully undertaken. In this study, we investigated the effect of diosgenin on the osteogenesis of murine MC3T3-E1 osteoblastic cells. Cells were cultured with varying levels of diosgenin (0-10 µM) within 25 days of bone formation period. Diosgenin was found to stimulate proliferation within the range of 0.01-5 µM using MTT assay. The medium and cellular levels of Type 1 collagen and alkaline phosphatase (ALP), both of which are major bone matrix proteins, increased within the low range of diosgenin concentration (>0-3 µM), and this pattern was further confirmed by collagen and ALP staining of the extracellular matrix (ECM). The cellular protein expression of ALP and collagen Type 1 was also increased at 0.1-1 µM diosgenin treatment as analyzed by Western blot. Calcium deposition within the ECM also showed the same pattern as assessed by Alizarin Red S and Von Kossa staining. Bone-specific transcription factor runt-related transcription factor 2 (Runx2) and Runx2-regulated osteopontin protein expressions were induced at low concentration (0.1-1 µM) and again decreased with high diosgenin concentrations. Based on our findings, our study suggests that diosgenin can enhance bone formation by stimulating the synthesis and secretion of Type 1 collagen and ALP and bone marker proteins Runx2 and osteopontin expression. The increased levels of these marker proteins, in turn, can increase the formation of calcium deposits within the ECM thereby increasing bone formation.