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1.
Technol Health Care ; 32(S1): 565-575, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38759077

RESUMO

BACKGROUND: Many people work in a sitting position where they have to use their upper extremities and hands. Muscle activity in sitting position is affected by the chair height of, the height of the worktable, and the distance to the working target. OBJECTIVE: The purpose of this study is to investigate differences in muscle activity between men and wemen at different chair heghts and working distances. METHODS: Surface electromyography (EMG) was employed to determine the muscle activity of the upper limbs and the trunk, while an arm-stretching task was performed in a sitting position. RESULTS: Women's muscle activity was higher than that of men in most muscles as the chair height and working distance changed. CONCLUSIONS: As the height of the chair or the working distance was changed, muscle activity was greater in women than in men for most of the muscles analyzed. Under the same conditions, women may feel more muscle fatigue than men, so it is necessary to consider gender in the working environment.


Assuntos
Eletromiografia , Músculo Esquelético , Postura Sentada , Humanos , Feminino , Masculino , Fatores Sexuais , Adulto , Músculo Esquelético/fisiologia , Comportamento Sedentário , Postura/fisiologia , Extremidade Superior/fisiologia , Adulto Jovem , Fadiga Muscular/fisiologia
2.
Exp Mol Med ; 55(7): 1506-1519, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37394590

RESUMO

Neurogenin 3 (NGN3) is a key transcription factor in the cell fate determination of endocrine progenitors (EPs) in the developing pancreas. Previous studies have shown that the stability and activity of NGN3 are regulated by phosphorylation. However, the role of NGN3 methylation is poorly understood. Here, we report that protein arginine methyltransferase-1 (PRMT1)-mediated arginine 65 methylation of NGN3 is required for the pancreatic endocrine development of human embryonic stem cells (hESCs) in vitro. We found that inducible PRMT1-knockout (P-iKO) hESCs did not differentiate from EPs into endocrine cells (ECs) in the presence of doxycycline. Loss of PRMT1 caused NGN3 accumulation in the cytoplasm of EPs and decreased the transcriptional activity of NGN3. We found that PRMT1 specifically methylates NGN3 arginine 65 and that this modification is a prerequisite for ubiquitin-mediated degradation. Our findings demonstrate that arginine 65 methylation of NGN3 is a key molecular switch in hESCs permitting their differentiation into pancreatic ECs.


Assuntos
Células-Tronco Embrionárias Humanas , Humanos , Células-Tronco Embrionárias Humanas/metabolismo , Metilação , Arginina/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Pâncreas , Diferenciação Celular/genética , Metiltransferases/metabolismo , Proteína-Arginina N-Metiltransferases/genética , Proteína-Arginina N-Metiltransferases/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo
3.
Mol Ther ; 31(5): 1480-1495, 2023 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-36932674

RESUMO

Optogenetic techniques permit non-invasive, spatiotemporal, and reversible modulation of cellular activities. Here, we report a novel optogenetic regulatory system for insulin secretion in human pluripotent stem cell (hPSC)-derived pancreatic islet-like organoids using monSTIM1 (monster-opto-Stromal interaction molecule 1), an ultra-light-sensitive OptoSTIM1 variant. The monSTIM1 transgene was incorporated at the AAVS1 locus in human embryonic stem cells (hESCs) by CRISPR-Cas9-mediated genome editing. Not only were we able to elicit light-induced intracellular Ca2+ concentration ([Ca2+]i) transients from the resulting homozygous monSTIM1+/+-hESCs, but we also successfully differentiated them into pancreatic islet-like organoids (PIOs). Upon light stimulation, the ß-cells in these monSTIM1+/+-PIOs displayed reversible and reproducible [Ca2+]i transient dynamics. Furthermore, in response to photoexcitation, they secreted human insulin. Light-responsive insulin secretion was similarly observed in monSTIM1+/+-PIOs produced from neonatal diabetes (ND) patient-derived induced pluripotent stem cells (iPSCs). Under LED illumination, monSTIM1+/+-PIO-transplanted diabetic mice produced human c-peptide. Collectively, we developed a cellular model for the optogenetic control of insulin secretion using hPSCs, with the potential to be applied to the amelioration of hyperglycemic disorders.


Assuntos
Diabetes Mellitus Experimental , Células-Tronco Pluripotentes Induzidas , Células Secretoras de Insulina , Ilhotas Pancreáticas , Células-Tronco Pluripotentes , Humanos , Camundongos , Animais , Secreção de Insulina , Diabetes Mellitus Experimental/terapia , Diabetes Mellitus Experimental/metabolismo , Ilhotas Pancreáticas/metabolismo , Células-Tronco Pluripotentes/metabolismo , Insulina/metabolismo , Células Secretoras de Insulina/metabolismo , Organoides , Células-Tronco Pluripotentes Induzidas/metabolismo , Diferenciação Celular
4.
Transl Res ; 258: 35-46, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-36805562

RESUMO

The objective of this study was to investigate whether CRISPR/Cas9-mediated suppression of A4GALT could rescue phenotype of Fabry disease nephropathy (FDN) using human induced pluripotent stem cells (hiPSCs) derived kidney organoid system. We generated FDN patient-derived hiPSC (CMC-Fb-002) and FD-specific hiPSCs (GLA-KO) by knock-out (KO) of GLA in wild-type (WT) hiPSCs using CRISPR/Cas9. We then performed A4GALT KO in both CMC-Fb-002 and GLA-KO to make Fb-002-A4GALT-KO and GLA/A4GALT-KO, respectively. Using these hiPSCs, we generated kidney organoids and compared alpha-galactosidase-A enzyme (α-GalA) activity, globotriaosylceramide (Gb-3) deposition, and zebra body formation under electron microscopy (EM). We also compared mRNA expression levels using RNA-seq and qPCR. Generated hiPSCs showed typical pluripotency markers without chromosomal disruption. Expression levels of GLA in CMC-Fb-002 and GLA-KO and expression levels of A4GALT in Fb-002-A4GALT-KO and GLA/A4GALT-KO were successfully decreased compared to those in WT-hiPSCs, respectively. Generated kidney organoids using these hiPSCs expressed typical nephron markers. In CMC-Fb-002 and GLA-KO organoids, α-GalA activity was significantly decreased along with increased deposition of Gb-3 in comparison with WT organoids. Intralysosomal inclusion body was also detected under EM. However, these disease phenotypes were rescued by KO of A4GALT in both GLA/A4GALT-KO and Fb-002-A4GALT-KO kidney organoids. RNA-seq showed increased expression levels of genes related to FDN progression in both GLA-mutant organoids compared to those in WT. Such increases were rescued in GLA/A4GALT-KO or Fb-002-A4GALT-KO organoids. CRISPR/Cas9 mediated suppression of A4GALT could rescue FDN phenotype. Hence, it can be proposed as a therapeutic approach to treat FDN.


Assuntos
Doença de Fabry , Células-Tronco Pluripotentes Induzidas , Nefropatias , Humanos , Doença de Fabry/genética , Doença de Fabry/metabolismo , Sistemas CRISPR-Cas/genética , Células-Tronco Pluripotentes Induzidas/metabolismo , Rim/metabolismo , Nefropatias/genética , Fenótipo , Organoides
5.
ACS Biomater Sci Eng ; 9(2): 680-692, 2023 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-36580628

RESUMO

Changes in the extracellular matrix (ECM) influence stem cell fate. When hESCs were differentiated on a thin layer of Matrigel coated onto PDMS (Matrigel_PDMS), they exhibited a substantial increase in focal adhesion and focal adhesion-associated proteins compared with those cultured on Matrigel coated onto TCPS (Matrigel_TCPS), resulting in YAP/TEF1 activation and ultimately promoting the transcriptional activities of pancreatic endoderm (PE)-associated genes. Interestingly, YAP activation in PE cells was mediated through integrin α3-FAK-CDC42-PP1A signaling rather than the typical Hippo signaling pathway. Furthermore, pancreatic islet-like organoids (PIOs) generated on Matrigel_PDMS secreted more insulin than those generated from Matrigel_TCPS. Electron micrographs revealed differential Matrigel architectures depending on the underlying substrate, resulting in varying cell-matrix anchorage resistance levels. Accordingly, the high apparent stiffness of the unique mucus-like network structure of Matrigel_PDMS was the critical factor that directly upregulated focal adhesion, thereby leading to better maturation of the pancreatic development of hESCs in vitro.


Assuntos
Células-Tronco Embrionárias Humanas , Humanos , Diferenciação Celular , Matriz Extracelular/metabolismo , Células-Tronco
6.
Artigo em Inglês | MEDLINE | ID: mdl-35805554

RESUMO

In South Korea, the suicide rate is more than double the OECD average, and precise identification of the cause is required for suicide prevention. Psychological autopsy is used to reveal factors related to suicidal behavior; however, such studies are lacking in Korea. This study investigated the factors related to suicide using psychological autopsies in Incheon, a major city in Korea. In total, 46 cases were investigated using the Korea-Psychological Autopsy Checklist (K-PAC), and data on mental health conditions and psychosocial factors of suicide decedents and their families were analyzed. It was estimated that 87% of individuals of suicides had a mental health condition before death, but only 15.2% continued treatment or counseling. Most individuals who died of suicide showed warning signs before death, but only 19.6% of survivors of suicide loss noticed them. Mental health concerns before and after the death of the individual were also identified in more than half of their families. To prevent suicide, intensive and continuous treatment for psychiatric conditions and prompt recognition of active response to suicide warning signs are required. Care for the mental health of family members is also important.


Assuntos
Suicídio , Autopsia , Humanos , República da Coreia/epidemiologia , Fatores de Risco , Ideação Suicida , Suicídio/psicologia , Sobreviventes
7.
Stem Cells ; 40(9): 870-882, 2022 09 26.
Artigo em Inglês | MEDLINE | ID: mdl-35852488

RESUMO

Allogeneic mesenchymal stem/stromal cells (MSCs) are frequently used in clinical trials due to their low expression of major histocompatibility complex (MHC) class I and lack of MHC class II. However, the levels of MHC classes I and II in MSCs are increased by inflammatory stimuli, raising concerns over potential adverse effects associated with allogeneic cell therapy. Also, it is unclear how the host immune response to MHC-mismatched MSCs affects the therapeutic efficacy of the cells. Herein, using strategies to manipulate MHC genes in human bone marrow-derived MSCs via the CRISPR-Cas9 system, plasmids, or siRNAs, we found that inhibition of MHC class I-not MHC class II-in MSCs lowered the survival rate of MSCs and their immunosuppressive potency in mice with experimental autoimmune uveoretinitis, specifically by increasing MSC vulnerability to natural killer (NK)-cell-mediated cytotoxicity. A subsequent survey of MSC batches derived from 6 human donors confirmed a significant correlation between MSC survival rate and susceptibility to NK cells with the potency of MSCs to increase MHC class I level upon stimulation. Our overall results demonstrate that MHC class I enables MSCs to evade NK-cell-mediated cytotoxicity and exert immunosuppressive activity.


Assuntos
Células-Tronco Mesenquimais , Animais , Antígenos HLA , Antígenos de Histocompatibilidade Classe II/genética , Antígenos de Histocompatibilidade Classe II/farmacologia , Humanos , Células Matadoras Naturais , Camundongos
8.
Exp Mol Med ; 54(6): 825-833, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35760874

RESUMO

Branched-chain aminotransferase 1 (BCAT1) transfers the amine group on branched-chain amino acids (BCAAs) to alpha-ketoglutarate. This generates glutamate along with alpha-keto acids that are eventually oxidized to provide the cell with energy. BCAT1 thus plays a critical role in sustaining BCAA concentrations and availability as an energy source. Osteoclasts have high metabolic needs during differentiation. When we assessed the levels of amino acids in bone marrow macrophages (BMMs) that were undergoing receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclast differentiation, we found that the BCAA levels steadily increase during this process. In vitro analyses then showed that all three BCAAs but especially valine were needed for osteoclast maturation. Moreover, selective inhibition of BCAT1 with gabapentin significantly reduced osteoclast maturation. Expression of enzymatically dead BCAT1 also abrogated osteoclast maturation. Importantly, gabapentin inhibited lipopolysaccharide (LPS)-induced bone loss of calvaria in mice. These findings suggest that BCAT1 could serve as a therapeutic target that dampens osteoclast formation.


Assuntos
Aminoácidos de Cadeia Ramificada , Osteoclastos , Transaminases/metabolismo , Aminoácidos de Cadeia Ramificada/metabolismo , Animais , Diferenciação Celular , Gabapentina/farmacologia , Camundongos , Osteoclastos/metabolismo , Ligante RANK/metabolismo
9.
J Gastrointest Surg ; 25(12): 3149-3159, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34244950

RESUMO

BACKGROUND OR PURPOSE: Pancreatic ductal adenocarcinoma (PDAC) is commonly diagnosed by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA). However, the diagnostic adequacy of EUS-FNA is often limited by low cellularity leading to inconclusive results. We aimed to investigate the feasibility and added utility of targeted next-generation sequencing (NGS) on PDAC EUS-FNAs. METHODS: EUS-FNAs were prospectively performed on 59 patients with suspected PDAC (2014-2017) at a high-volume center. FNAs were analyzed for the presence of somatic mutations using NGS to supplement cytopathologic evaluations and were compared to surgical specimens and circulating tumor DNA (ctDNA). RESULTS: Fifty-nine patients with suspected PDAC were evaluated, and 52 were diagnosed with PDAC on EUS-FNA. Four of the remaining seven patients had inconclusive EUS-FNAs and were ultimately diagnosed with PDAC after surgical resection. Of these 56 cases of PDAC, 48 (85.7%) and 18 (32.1%) harbored a KRAS and/or TP53 mutation on FNA NGS, respectively. Particularly, in the four inconclusive FNA PDAC diagnoses (false negatives), half harbored KRAS mutations on FNA. No KRAS/TP53 mutation was found in remaining three non-PDAC cases. All EUS-FNA detected KRAS mutations were detected in 16 patients that underwent primary tumor NGS (100% concordance), while 75% KRAS concordance was found between FNA and ctDNA NGS. CONCLUSION: Targeted NGS can reliably detect KRAS mutations from EUS-FNA samples and exhibits high KRAS mutational concordance with primary tumor and ctDNA. This suggests targeted NGS of EUS-FNA samples may enable preoperative ctDNA prognostication using digital droplet PCR and supplement diagnoses in patients with inconclusive EUS-FNA.


Assuntos
Endossonografia , Neoplasias Pancreáticas , Aspiração por Agulha Fina Guiada por Ultrassom Endoscópico , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Mutação , Neoplasias Pancreáticas/diagnóstico por imagem , Neoplasias Pancreáticas/genética , Estudos Prospectivos
10.
J Med Internet Res ; 23(2): e20177, 2021 02 16.
Artigo em Inglês | MEDLINE | ID: mdl-33591276

RESUMO

BACKGROUND: Metamemory training (MMT) is a useful training strategy for improving cognitive functioning in the older adult population. Despite the advantages, there are limitations imposed by location and time constraints. OBJECTIVE: This study aimed to develop a smart speaker-based MMT program and evaluate the efficacy of the program in older adults without cognitive impairment. METHODS: This study used a case-control cohort design. The smart speaker-based MMT program comprised 3 training sessions per day, 5 days a week, for 8 weeks. Each training session took approximately 15 minutes. This program was implemented using smart speakers, not human trainers. All participants completed the Mini-Mental State Examination, Subjective Memory Complaints Questionnaire, Verbal Learning Test, Digit Span Test, fluency tests, and a short-form version of the Geriatric Depression Scale before and after training. RESULTS: A total of 60 subjects (29 in the MMT group and 31 in the control group) participated in the study. The training group showed significant increases in the delayed free recall, digit span forward, digit span backward, and fluency test scores compared with the control group. CONCLUSIONS: This study confirmed the efficacy of smart speaker-based MMT in older adults. Home-based smart speaker-based MMT is not limited with respect to location or constrained by space and may help older adults with subjective cognitive decline without requiring intervention by human professionals.


Assuntos
Inteligência Artificial/tendências , Metacognição/fisiologia , Ensino/normas , Estudos de Casos e Controles , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
11.
J Vasc Interv Radiol ; 31(9): 1483-1491, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32800664

RESUMO

PURPOSE: To examine safety and efficacy of bariatric arterial embolization (BAE) with x-ray-visible embolic microspheres (XEMs) and an antireflux catheter in swine. MATERIAL AND METHODS: BAE with selective infusion of XEMs (n = 6) or saline (n = 4, control) into gastric fundal arteries was performed under x-ray guidance. Weight and plasma hormone levels were measured at baseline and weekly for 4 weeks after embolization. Cone-beam CT images were acquired immediately after embolization and weekly for 4 weeks. Hormone-expressing cells in the stomach were assessed by immunohistochemical staining. RESULTS: BAE pigs lost weight 1 week after embolization followed by significantly impaired weight gain relative to control animals (14.3% vs 20.9% at 4 weeks, P = .03). Plasma ghrelin levels were significantly lower in BAE pigs than in control animals (1,221.6 pg/mL vs 1,706.2 pg/mL at 4 weeks, P < .01). XEMs were visible on x-ray and cone-beam CT during embolization, and radiopacity persisted over 4 weeks (165.5 HU at week 1 vs 158.5 HU at week 4, P = .9). Superficial mucosal ulcerations were noted in 1 of 6 BAE animals. Ghrelin-expressing cell counts were significantly lower in the gastric fundus (17.7 vs 36.8, P < .00001) and antrum (24.2 vs 46.3, P < .0001) of BAE pigs compared with control animals. Gastrin-expressing cell counts were markedly reduced in BAE pigs relative to control animals (98.5 vs 127.0, P < .02). Trichrome staining demonstrated significantly more fibrosis in BAE animals compared with control animals (13.8% vs 8.7%, P < .0001). CONCLUSIONS: XEMs enabled direct visualization of embolic material during and after embolization. BAE with XEMs and antireflux microcatheters was safe and effective.


Assuntos
Regulação do Apetite , Comportamento Animal , Catéteres , Embolização Terapêutica/instrumentação , Artéria Gástrica , Fundo Gástrico/irrigação sanguínea , Grelina/sangue , Redução de Peso , Animais , Tomografia Computadorizada de Feixe Cônico , Artéria Gástrica/diagnóstico por imagem , Fundo Gástrico/metabolismo , Fundo Gástrico/patologia , Infusões Intra-Arteriais , Microesferas , Sus scrofa , Fatores de Tempo
12.
Prev Nutr Food Sci ; 25(1): 78-83, 2020 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-32292759

RESUMO

In this study, the antioxidant effects of seven types of commercial Chunjang from China (C1∼3) and Korea (K1∼4) were compared for their ability to protect against H2O2-induced DNA damage. Outputs included total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (DPPH RSA), oxygen radical absorbance capacity (ORAC), total radical trapping antioxidant potential (TRAP), and comet assays. TPC was the highest in C3 (1,250.8 mg gallic acid equivalent/100 g). C1 exhibited a significantly higher DPPH RSA IC50 (7.5 mg/mL) and ORAC concentration (8.22 µM Trolox equivalent) compared with all other samples, and C1 and K1 exhibited the highest TRAPs. H2O2-induced DNA damage was effectively protected by Chunjang, with a higher observed in C2, C3, and K1 (24.2∼25.3 µg/mL) compared with the other samples (28.3∼30.0 µg/mL). Our results showed that commercial Chunjang contains polyphenol and antioxidant activities. The differences between the samples might be attributed to different origins, materials, and processing methods.

13.
Sci Rep ; 9(1): 17197, 2019 11 20.
Artigo em Inglês | MEDLINE | ID: mdl-31748571

RESUMO

Large DNA molecules are a promising platform for in vitro single-molecule biochemical analysis to investigate DNA-protein interactions by fluorescence microscopy. For many studies, intercalating fluorescent dyes have been primary DNA staining reagents, but they often cause photo-induced DNA breakage as well as structural deformation. As a solution, we previously developed several fluorescent-protein DNA-binding peptides or proteins (FP-DBP) for reversibly staining DNA molecules without structural deformation or photo-induced damage. However, they cannot stain DNA in a condition similar to a physiological salt concentration that most biochemical reactions require. Given these concerns, here we developed a salt-tolerant FP-DBP: truncated transcription activator-like effector (tTALE-FP), which can stain DNA up to 100 mM NaCl. Moreover, we found an interesting phenomenon that the tTALE-FP stained DNA evenly in 1 × TE buffer but showed AT-rich specific patterns from 40 mM to 100 mM NaCl. Using an assay based on fluorescence resonance energy transfer, we demonstrated that this binding pattern is caused by a higher DNA binding affinity of tTALE-FP for AT-rich compared to GC-rich regions. Finally, we used tTALE-FP in a single molecule fluorescence assay to monitor real-time restriction enzyme digestion of single DNA molecules. Altogether, our results demonstrate that this protein can provide a useful alternative as a DNA stain over intercalators.


Assuntos
Proteínas de Ligação a DNA/metabolismo , DNA/química , DNA/metabolismo , Corantes Fluorescentes/química , Substâncias Intercalantes/metabolismo , Coloração e Rotulagem/métodos , Efetores Semelhantes a Ativadores de Transcrição/metabolismo , Proteínas de Ligação a DNA/química , Fluorescência , Transferência Ressonante de Energia de Fluorescência , Humanos , Substâncias Intercalantes/química , Microscopia de Fluorescência , Imagem Individual de Molécula/métodos , Efetores Semelhantes a Ativadores de Transcrição/química
14.
Adv Exp Med Biol ; 1155: 567-581, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468432

RESUMO

Here, we investigated the hepatoprotective effect of a hot water extract from Loliolus beka gray meat (LBMH) containing plentiful taurine in H2O2-induced oxidative stress in hepatocytes. LBMH potently scavenged the 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals and exhibited the good reducing power and the oxygen radical absorbance capacity (ORAC) value. Also, LBMH improved the cell viability against H2O2-induced hepatic damage in cultured hepatocytes by reducing intracellular reactive oxygen species (ROS) production. In addition, LBMH inhibited apoptosis via a reduction in sub-G1 cell population, as well as inhibition of apoptotic body formation from H2O2-induced oxidative damage in hepatocytes. Moreover, LBMH regulated the expression levels of Bax, a pro-apoptotic molecule and Bcl-2, an anti-apoptotic molecule in H2O2-treated hepatocytes. Additionally, pre-treatment with LBMH increased the expression of heme oxygenase 1 (HO-1), which is a hepatoprotective enzyme, by activating the nuclear factor erythroid 2-related factor 2 (Nrf2) in H2O2-treated hepatocytes. Taken together, LBMH may be useful as a food ingredient for treatment of liver disease by regulating the Nrf2/HO-1 signal pathway.


Assuntos
Antioxidantes , Extratos Celulares/farmacologia , Decapodiformes/química , Hepatócitos/efeitos dos fármacos , Estresse Oxidativo , Taurina/farmacologia , Animais , Células Cultivadas , Heme Oxigenase-1/metabolismo , Hepatócitos/citologia , Humanos , Peróxido de Hidrogênio , Carne , Fator 2 Relacionado a NF-E2/metabolismo , Espécies Reativas de Oxigênio/metabolismo
15.
Adv Exp Med Biol ; 1155: 583-596, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468433

RESUMO

In this study, we evaluated the protective effects of an aqueous extract from Batillus cornutus meat (BM) against cellular oxidative damage caused by hydrogen peroxide (H2O2) in human hepatocyte, Chang cells. First, we prepared an aqueous extract of BM meat (BMW) showing the highest taurine content among free amino acid contents. BMW led to high antioxidant activity showing 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radical scavenging activity, good reducing power and an oxygen radical absorbance capacity (ORAC) value. Also, BMW improved cell viability that was diminished by H2O2 exposure, as it reduced the generation of intracellular reactive oxygen species (ROS) in Chang cells. In addition, BMW up-regulated the production of antioxidant enzymes, such as catalase and superoxide dismutase (SOD), compared to H2O2-treated Chang cells lacking BMW. Moreover, BMW induced the expressions of nuclear Nrf2 and cytosolic HO-1 in H2O2-treated Chang cells. Interestingly, the treatment of ZnPP, HO-1 inhibitor, abolished the improvement in cell viability and intracellular ROS generation mediated by BMW treatment. In conclusion, this study suggests that BMW protects hepatocytes against H2O2-mediated cellular oxidative damage via up-regulation of the Nrf2/HO-1 signal pathway.


Assuntos
Extratos Celulares/farmacologia , Gastrópodes/química , Heme Oxigenase-1/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo , Transdução de Sinais , Taurina/farmacologia , Animais , Células Cultivadas , Humanos , Peróxido de Hidrogênio , Carne , Espécies Reativas de Oxigênio/metabolismo , Regulação para Cima
16.
Adv Exp Med Biol ; 1155: 597-610, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468434

RESUMO

Octopus ocellatus meat (OM) is well known as a plentiful protein source. In this study, we evaluated the hepatoprotective effect of an aqueous extract of OM (OMA) against H2O2-triggered oxidative stress in human hepatocytes. First of all, taurine rich OMA showed a good ORAC value and reducing power and it was similar with that of ascorbic acid, which is known as a strong antioxidant. Also, OMA significantly improved H2O2-decreased cell viability by reducing the generation of intracellular reactive oxygen species (ROS) in hepatocytes. Interestingly, the stimulation of H2O2-induced the formations of apoptotic bodies and sub-G1 DNA content, whereas they were inhibited by the treatment with OMA. Furthermore, OMA regulated the protein expression levels of apoptotic molecules, such as Bax and Bcl-2. Taken together, this study suggests that OMA, which contains an abundant amount of taurine, protects hepatocytes from H2O2-triggered oxidative stress and might be a functional food material with hepatoprotective effects.


Assuntos
Extratos Celulares/farmacologia , Hepatócitos/efeitos dos fármacos , Octopodiformes/química , Estresse Oxidativo , Animais , Apoptose , Células Cultivadas , Hepatócitos/citologia , Humanos , Peróxido de Hidrogênio/efeitos adversos , Carne , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína X Associada a bcl-2/metabolismo
17.
Adv Exp Med Biol ; 1155: 643-659, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468437

RESUMO

Batillus cornutus (B. cornutus) is one of the gastropoda, which are distributed along the coast of China, Japan and South Korea and northeast area. In this study, we first identified the antioxidant effects of a B. cornutus meat (BM) enzymatic hydrolysate in H2O2-treated Vero cells. First of all, we prepared an Alcalase hydrolysate from BM (BMA) and revealed a high taurine content. Also, taurine rich BMA dose-dependently increased 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radical scavenging activity, reducing power and the higher oxygen radical absorbance capacity (ORAC) value. In addition, BMA significantly increased the cell viability via the down-regulation of intracellular reactive oxygen species (ROS) production, as well as the decreased formation of apoptotic bodies and sub-G1 DNA population in H2O2-treated Vero cells. Furthermore, BMA increased the expression of the anti-apoptotic molecule, Bcl-2, and decreased the expressions of Bax, p53 and cleaved PARP, all of which are pro-apoptotic molecules, in H2O2-treated Vero cells. Based on these results, this study suggests that BMA may be used as a potential protector on damage caused by oxidative stress.


Assuntos
Antioxidantes/farmacologia , Gastrópodes/química , Estresse Oxidativo , Hidrolisados de Proteína/farmacologia , Animais , Chlorocebus aethiops , Peróxido de Hidrogênio , Carne , Espécies Reativas de Oxigênio/metabolismo , Subtilisinas , Células Vero
18.
Adv Exp Med Biol ; 1155: 661-674, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468438

RESUMO

In the present study, we investigated the antioxidant activity of an aqueous extract from Atrina pectinate meat (APW) against H2O2-induced oxidative stress in a human hepatocyte. The extraction yield of APW was 30.01 ± 0.83% and which contained the highest taurine content among free amino acid contents. APW led to the high antioxidant activity showing 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radical scavenging activity, good reducing power and oxygen radical absorbance capacity (ORAC) value. Also, the results showed that APW improved the cell viability decreased by H2O2 stimulation as well as the reduction of intracellular reactive oxygen species (ROS) generation in hepatocytes. Additionally, APW up-regulated the production of antioxidant mechanisms related enzymes such as catalase and superoxide dismutase (SOD), compared to the only H2O2-treated hepatocytes. Moreover, APW increased the expressions of nuclear Nrf2 and cytosolic HO-1 in H2O2-treated hepatocytes. Interestingly, the treatment of ZnPP, a HO-1 inhibitor abolished the cell viability and intracellular ROS generation induced by APW treatment. In conclusion, this study suggests that APW protects H2O2 induced oxidative stress via up-regulating of Nrf2/HO-1 signal pathway in hepatocytes.


Assuntos
Antioxidantes/farmacologia , Bivalves/química , Extratos Celulares/farmacologia , Hepatócitos/efeitos dos fármacos , Estresse Oxidativo , Animais , Heme Oxigenase-1/metabolismo , Humanos , Peróxido de Hidrogênio , Carne , Fator 2 Relacionado a NF-E2/metabolismo , Espécies Reativas de Oxigênio/metabolismo
19.
Adv Exp Med Biol ; 1155: 691-703, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468440

RESUMO

In this study, we investigated the hepatoprotective activity of the water extract derived from Octopus vulgaris meat (OM). First of all, a water extract prepared from OM (OMW) showed the high extraction yield (48.22%) and the highest taurine content (39.84%) in free amino acids. OMW exhibited the high value of reducing power, ABTS and hydrogen peroxide radical scavenging activities in dose-dependent manner. The taurine-rich OMW also led to the reduced intracellular reactive oxygen species (ROS) generation with the increased cell survival in H2O2-treated Chang liver cells. In addition, OMW decreased the apoptotic phenomenon, including the formations of apoptotic bodies and sub-G1 DNA contents by regulating the protein expressions of apoptosis-related molecules such as Bcl-2 and Bax. From these results, this study indicated the taurine-rich OMW protected hepatocytes against oxidative stress. These findings suggest that OWM may be a novel potential antioxidant resource.


Assuntos
Extratos Celulares/farmacologia , Hepatócitos/citologia , Octopodiformes/química , Estresse Oxidativo , Taurina/farmacologia , Animais , Antioxidantes/farmacologia , Linhagem Celular , Humanos , Peróxido de Hidrogênio , Carne , Espécies Reativas de Oxigênio , Água
20.
Adv Exp Med Biol ; 1155: 705-715, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468441

RESUMO

Blood vessels become less flexible with senescence; arteries narrow and become less flexible, disturbing blood circulation in aging and other vascular diseases. Mechanistically, vascular senescence plays an important role in the pathogenesis of normal aging and age-related vascular diseases. Vascular senescence also causes vascular dysfunction, resulting in damage to the vessel wall. Vascular aging involves the senescence of endothelial cells. Hydrogen peroxide is widely used to achieve oxidative stress-induced premature senescence. Here, we investigated the protective effects of a hot water extract of Loliolus beka meat (LBM) against H2O2-exposed HUVECs, a human umbilical vein endothelial cells line. The hot water extract of LBM protected cells against H2O2-induced cytotoxicity while reducing the expression of senescence markers, including ß-galactosidase, p53, and p21. In addition, the hot water extract of LBM protected against H2O2-induced DNA damage. These findings suggest that the hot water extract of LBM protects HUVECs from H2O2-induced senescence by preventing cellular damage. LBM serve as a supplement or natural food with benefits against vascular disease.


Assuntos
Extratos Celulares/farmacologia , Decapodiformes/química , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Estresse Oxidativo , Animais , Células Cultivadas , Senescência Celular , Humanos , Peróxido de Hidrogênio , Carne
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