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1.
Crit Rev Food Sci Nutr ; 62(25): 6908-6931, 2022.
Article in English | MEDLINE | ID: mdl-33819119

ABSTRACT

ß-glucans, the class of biological response modifier has unceasing attention, not only for its immune stimulating but also for its role as prebiotics, modulator of physiological events etc. and is widely used in the treatment of cancer, diabetes, gastrointestinal disorders, cardiovascular diseases etc. However, ß-glucan with different physiochemical properties is found to have discrete clinical functions and thus careful selection of the types of ß-glucan plays pivotal role in providing significant and expected clinical outcome. Herein this review, we presented the factors responsible for diverse functional properties of ß-glucan, their distinct mode of actions in regulating human health etc. Further, clinical aspects of different ß-glucans toward the management of wound care, metabolic dysbiosis, fatty liver disorders and endurance training associated energy metabolism were compiled and exhibited in detail.


Subject(s)
beta-Glucans , Humans , Immunologic Factors , Prebiotics , beta-Glucans/chemistry
2.
Eur J Nutr ; 61(2): 793-807, 2022 Mar.
Article in English | MEDLINE | ID: mdl-34561722

ABSTRACT

PURPOSE: Probiotics and prebiotics are commonly used to improve the gut microbiota. Since prebiotics can support the growth of probiotics, co-administration of these is called synbiotics. It has been demonstrated that obesity-induced gut dysbiosis can worsen inflammatory bowel disease symptoms. This study evaluated how modulation of gut microbiota with Schizophyllum commune-derived ß-glucan (BG), probiotics (PRO), and synbiotics containing both BG and PRO (SYN) could improve the symptoms of obesity-associated colitis and hepatic manifestation. METHODS: Mice were fed a normal diet (ND), high-fat diet (HFD), and HFD with different additives (BG, PRO, and SYN) for 12 weeks, followed by 5 days of colitis induction. Mice were sacrificed before and after colitis induction. During the experiment, body weight, food and water consumption, and rectal bleeding were monitored. Proteins from the colon were subjected to western blotting, and serum biomarkers such as alanine transaminase, alkaline phosphatase, triglycerides, and total cholesterol were analyzed. Colon and liver samples were sectioned for histological analysis. The fecal microbiota was analyzed based on partial 16S rRNA gene sequences. RESULTS: Although BG and PRO secured intestinal tight junctions, these two treatments did not modulate inflammatory cell infiltration and inflammatory markers (i.e., IL-6 and TNF-α). In contrast, SYN demonstrated stronger and broader effects in reducing colonic inflammation. While BG treatment increased the abundance of indigenous Lactobacillus, PRO treatment decreased bacterial diversity by suppressing the growth of several species of bacteria. SYN treatment groups, however, supported the growth of both indigenous and supplemented bacteria while maintaining bacterial diversity. CONCLUSION: Obesity-associated colitis can be improved by modulating gut bacteria with ß-glucan and probiotics. The co-administration of both outperformed ß-glucan and probiotic treatment alone by fostering both indigenous and supplemented probiotic strains.


Subject(s)
Colitis , Probiotics , Synbiotics , beta-Glucans , Animals , Colitis/metabolism , Liver/metabolism , Mice , Mice, Inbred C57BL , Obesity/metabolism , Probiotics/pharmacology , RNA, Ribosomal, 16S/genetics , beta-Glucans/pharmacology
5.
Arthritis Res Ther ; 22(1): 275, 2020 11 23.
Article in English | MEDLINE | ID: mdl-33228785

ABSTRACT

BACKGROUND: Interleukin-6 (IL-6) is involved in fibroblast-like synoviocyte (FLS) activation and promotes pannus formation and bone and cartilage destruction in rheumatoid arthritis (RA). Cysteine-rich 61 (Cyr61) protein regulates cell proliferation, migration, and differentiation. The aim of this study was to investigate the role of Cyr61 in RA-FLS migration and invasion after IL-6 stimulation. METHODS: Western blotting, immunohistochemistry, reverse transcription-polymerase chain reaction, and real time-polymerase chain reaction were used to examine protein and mRNA levels of Cyr61, matrix metalloproteinases (MMPs), and other signalling proteins. Knockdown of gene expression was performed with siRNA, and RNA sequencing was performed for differential gene analysis. Migration and invasion were assessed by wound healing and Boyden chamber assays. RESULTS: Cyr61 levels were elevated in FLSs from RA patients compared to those in osteoarthritis patients. Control and IL-6-treated FLSs showed differential gene expression. IL-6 stimulated protein synthesis of Cyr61, which was attenuated by the extracellular signal-related kinase 1/2 (ERK 1/2) inhibitor, PD98059, and knockdown of early growth response 3 (EGR3), but not of JUN. IL-6-induced Cyr61 protein synthesis increased expression of MMP2. Cyr61 promoted FLS migration and invasion in an autocrine manner. Knockdown of CYR61 and a neutralising antibody attenuated Cyr61 synthesis and IL-6-induced FLS migration. CONCLUSIONS: By modulating the ERK/EGR3 pathway, IL-6 stimulated Cyr61 production and in turn increased invasiveness of FLS. Our data suggest that Cyr61 might be a potential target to prevent the progression of joint damage in RA.


Subject(s)
Arthritis, Rheumatoid , Interleukin-6 , Synoviocytes , Cell Movement , Cells, Cultured , Cysteine-Rich Protein 61/genetics , Fibroblasts , Humans , Interleukin-6/physiology , Synovial Membrane
6.
Cells ; 9(6)2020 06 26.
Article in English | MEDLINE | ID: mdl-32604782

ABSTRACT

Epithelial-to-mesenchymal transition (EMT) promulgates epithelial cell associated disease-defining characteristics in tumorigenesis and organ fibrosis. Growth factors such as epidermal growth factor and fibroblast growth factor in addition to cytokines such as transforming growth factor-ß1 (TGF-ß1) is said to play a prominent role in remodeling related pathological events of cancer progression such as invasion, metastasis, apoptosis, EMT, etc. through redox related cellular secondary messengers, in particular the reactive oxygen species (ROS). However, the signaling cascade underlying the redox mechanism and thereby the progression of EMT remains largely unknown. In this study, upon TGF-ß1 treatment, we observed an induction in NOX isoforms-NOX2 and NOX4-that have time (early and late) and cellular localization (nucleus and autophagosome co-localized) dependent effects in mediating EMT associated cell proliferation and migration through activation of the focal adhesion kinase (FAK)/SRC pathway in HeLa, human cervical cancer cells. Upon silencing NOX2/4 gene expression and using the SRC inhibitor (AZD0530), progression of TGF-ß1 induced EMT related cellular remodeling, extra cellular matrix (ECM) production, cell migration and invasion, got significantly reverted. Together, these results indicate that NOX2 and NOX4 play important, albeit distinct, roles in the activation of cytokine mediated EMT and its associated processes via tyrosine phosphorylation of the FAK/SRC pathway.


Subject(s)
Focal Adhesion Kinase 1/genetics , NADPH Oxidases/metabolism , Protein Isoforms/metabolism , Uterine Cervical Neoplasms/genetics , Epithelial Cells , Epithelial-Mesenchymal Transition , Female , HeLa Cells , Humans , Oxidation-Reduction , Phosphorylation , Transfection
7.
J Cancer ; 11(15): 4495-4509, 2020.
Article in English | MEDLINE | ID: mdl-32489467

ABSTRACT

Objective: We propose that sirtuin (SIRT) may induce a pro-apoptotic effect by deacetylating transcription factors in A549 cells: depletion of sirtuin-1 (SIRT1) induced cell cycle arrest in cisplatin-resistant A549 (A549/CADD) cells. Methods: Protein and mRNA levels of SIRT1 were investigated using western blot and RT-PCR. In A549 and A549/CADD cells, the cytotoxicity of cisplatin administration was evaluated by MTT assay, proliferation was measured by ECIS, and the cell cycle distribution was analyzed using FACS. Cells were transfected with pcDNA3.1-Myc-SIRT1 or pcDNA3.1-Myc-Control vectors to analyze the impact of SIRT-1 on cisplatin induced drug resistance. SIRT1 localization was studied using immunofluorescence analysis. In addition, immunoprecipitation and 20S proteasome activity assay were performed to examine the relationship of SIRT1 with the proteasome complex. Results: A549/CADD cells exhibited a mesenchymal-like cell characteristic. SIRT1 expression was markedly decreased in A549/CADD cells. We observed that cisplatin regulates p53 stability through the depletion of ubiquitination following SIRT1 downregulation. Furthermore, cisplatin treatment increased proteasomal activity and significantly decreased cytoplasmic SIRT1 protein levels in A549/CADD cells. Conclusion: In this study, we found SIRT1 to be depleted in A549/CADD cells and also determined the underlying resistance mechanism which may act as novel therapeutic targets in overcoming drug resistance.

8.
Food Sci Nutr ; 8(4): 2059-2067, 2020 Apr.
Article in English | MEDLINE | ID: mdl-32328272

ABSTRACT

Myricetin is a commonly found dietary flavonoid. In the present study, we investigated the effects of myricetin on migration and invasion of radioresistant lung cancer cells (A549-IR). Transcriptome analysis of A549-IR cells identified several differentially expressed genes (DEGs) in A549-IR cells compared to parental A549 cells. Functional enrichment analysis revealed that most of the DEGs were linked with PI3K-AKT signaling, proteoglycans, focal adhesion, and ECM-receptor interactions. A549-IR cells demonstrated enhanced migratory potential with increased expression of vimentin, snail and slug, and reduced expression of E-cadherin. A549-IR cells exposed to myricetin displayed reduced migration and suppressed MMP-2 and MMP-9 expression. Notably, myricetin inhibited the phosphorylation of focal adhesion kinase (FAK) and altered the F-actin/G-actin ratio in A549-IR cells, without modulation of EMT markers. These findings suggest that myricetin can inhibit migration of A549-IR cells by suppressing MMP-2 and MMP-9 expressions through inhibition of the FAK-ERK signaling pathway.

9.
Cell Biol Int ; 44(2): 477-487, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31631444

ABSTRACT

With poor prognosis and aberrant lung remodeling, pulmonary fibrosis exhibits worldwide prevalence accompanied by an increase in burden in terms of hospitalization and death. Apart from genetic and non-genetic factors, fibrosis occurs as a side effect of bleomycin antineoplastic activity. Elucidating the cellular and molecular mechanism could help in the development of effective anti-fibrotic treatment strategies. In the present study, we investigated the underlying mechanism behind bleomycin-induced fibrosis using human alveolar epithelial cells (A549 cells). On the basis of the experimental observation, it was demonstrated that with transforming growth factor-ß (TGF-ß) as a central mediator of fibrosis progression, a cross-talk between epithelial-mesenchymal transition (EMT) and senescence upon bleomycin treatment occurs. This results in the advancement of this serious fibrotic condition. Fibrosis was initiated through integrin activation and imbalance in the redox state (NOX expression) of the cell. It progressed along the TGF-ß-mediated non-canonical pathway (via ERK phosphorylation) followed by the upregulation of α-smooth muscle actin and collagen synthesis. Additionally, in this process, the loss of the epithelial marker E-cadherin was observed. Furthermore, the expressions of senescence markers, such as p21 and p53, were upregulated upon bleomycin treatment, thereby intensifying the fibrotic condition. Accordingly, the molecular pathway mediating the bleomycin-induced fibrosis was explored in the current study.


Subject(s)
Apoptosis , Bleomycin/pharmacology , Cell Proliferation , Cellular Senescence , Epithelial-Mesenchymal Transition/drug effects , Pulmonary Fibrosis/pathology , Transforming Growth Factor beta/metabolism , A549 Cells , Antibiotics, Antineoplastic/pharmacology , Cell Movement , Humans , Pulmonary Fibrosis/chemically induced , Pulmonary Fibrosis/metabolism , Transforming Growth Factor beta/genetics
10.
Eur J Nutr ; 59(6): 2617-2629, 2020 Sep.
Article in English | MEDLINE | ID: mdl-31664519

ABSTRACT

PURPOSE: Western diet, rich in carbohydrates and fat, is said to be a major factor underlying metabolic syndrome. Interventions with prebiotics, the key modulators of the gut microbiota, have paramount impact on host-associated metabolic disorders. Herein, we investigated the effect of fungus-derived (1,3)/(1,6)-ß-glucan, a highly soluble dietary fiber, on high-fat diet (HFD)-induced metabolic distress. METHODS: Male C57BL/6 J mice were fed with different diet groups (n = 11): control diet, HFD, 3 g/kg or 5 g/kg of ß-glucan-incorporated HFD. At the end of experimental study period (12th week), body weight, feces weight and fecal moisture content were observed. Further, colonic motility was measured using activated charcoal meal study. Proteins extracted from liver and intestine tissues were subjected to western blot technique. Paraffin-embedded intestinal tissues were sectioned for histochemical [Periodic acid-Schiff (PAS) and Alcian blue (AB) staining] analysis. Fecal microbiota analysis was performed using MOTHUR bioinformatic software. RESULTS: ß-glucan consumption exhibited anti-obesity property in mice groups fed with HFD. In addition, ß-glucan ameliorated HFD-induced hepatic stress, colonic motility and intestinal atrophy (reduction in colon length, goblet cells, and mucosal layer thickness). Further, ß-glucan incorporation shifted bacterial community by increasing butyrate-producing bacteria such as Anaerostipes, Coprobacillus, and Roseburia and decreasing reportedly obesity-associated bacteria such as Parabacteroides and Lactococcus. CONCLUSION: Altogether, the outcomes of this present pre-clinical animal study show ß-glucan to be a promising therapeutic candidate in the treatment of HFD-induced metabolic distress. Further comprehensive research has to be conducted to brace its clinical relevance, reproducibility and efficacy for aiding human health.


Subject(s)
Gastrointestinal Microbiome , beta-Glucans , Animals , Diet, High-Fat/adverse effects , Fungi , Male , Mice , Mice, Inbred C57BL , Prebiotics , Reproducibility of Results
11.
Adv Wound Care (New Rochelle) ; 8(4): 125-135, 2019 Apr 01.
Article in English | MEDLINE | ID: mdl-31737411

ABSTRACT

Objective: Recognized as pathogen-associated molecular patterns (PAMPs), ß-glucans, a naturally occurring heterogeneous group of polysaccharides, were investigated for their ability to accelerate wound healing in the form of high water-retaining hydrogel dressing. Approach: Full-thickness wounds on the dorsal side of mice created using a 5-mm biopsy punch were treated with ß-glucan-based hydrogel for 2 weeks. Standardized photographs of the wound site were taken at regular time intervals to calculate the percentage of wound closure. Tissues isolated from the wound area were subjected to histological examination and immunoblot analysis. Results: ß-Glucan-based hydrogel significantly accelerated the duration of wound healing and enhanced the development of skin appendages in the regenerated skin tissue. Increased expression of transforming growth factor-ß3 in the skin tissue isolated from the healed wound site indicated that skin regeneration rather than skin repair occurred, thereby minimizing cutaneous scarring. The expression level of cytokeratin 10 and cytokeratin 14 in the isolated skin tissue revealed that the wounds treated with hydrogel showed proper differentiation and proliferation of keratinocytes in the epidermal layer. Innovation: Immunomodulating ß-glucan (responsible for fighting infections at the wound site, and enhancing the migration and proliferation of keratinocytes and fibroblasts) in the form of a three-dimensional hydrogel membrane that retains a high water content (responsible for cooling and soothing effect around the wound site, thereby reducing pain) was prepared and analyzed for its effects on the cutaneous wound healing mechanism. Conclusion: ß-Glucan-based hydrogels are promising as wet wound dressings in the health care industry.

12.
Cell Death Dis ; 10(9): 645, 2019 09 09.
Article in English | MEDLINE | ID: mdl-31501417

ABSTRACT

Reactive oxygen species (ROS) regulate cell fate, although signaling molecules that regulate ROS hormesis remain unclear. Here we show that transmembrane 4 L six family member 5 (TM4SF5) in lung epithelial cells induced the alternatively spliced CD44v8-10 variant via an inverse ZEB2/epithelial splicing regulatory proteins (ESRPs) linkage. TM4SF5 formed complexes with the cystine/glutamate antiporter system via TM4SF5- and CD44v8-10-dependent CD98hc plasma-membrane enrichment. Dynamic TM4SF5 binding to CD98hc required CD44v8-10 under ROS-generating inflammatory conditions. TM4SF5 and CD44v8-10 upregulated cystine/glutamate antiporter activity and intracellular glutathione levels, leading to ROS modulation for cell survival. Tm4sf5-null mice exhibited attenuated bleomycin-induced pulmonary fibrosis with lower CD44v8-10 and ESRPs levels than wild-type mice. Primary mouse alveolar epithelial cells (AECs) revealed type II AECs (AECII), but not type I, to adapt the TM4SF5-mediated characteristics, suggesting TM4SF5-mediated AECII survival following AECI injury during idiopathic pulmonary fibrosis (IPF). Thus, the TM4SF5-mediated CD44v8-10 splice variant could be targeted against IPF.


Subject(s)
Alveolar Epithelial Cells/metabolism , Hyaluronan Receptors/metabolism , Membrane Proteins/metabolism , Pulmonary Fibrosis/metabolism , A549 Cells , Alveolar Epithelial Cells/pathology , Animals , Cell Line, Tumor , Cell Survival/physiology , Humans , Hyaluronan Receptors/genetics , Male , Membrane Proteins/genetics , Mice, Inbred C57BL , Mice, Knockout , Pulmonary Fibrosis/genetics , Pulmonary Fibrosis/pathology , RNA Splicing , Reactive Oxygen Species/metabolism
13.
Int J Mol Sci ; 19(4)2018 Apr 11.
Article in English | MEDLINE | ID: mdl-29641498

ABSTRACT

Cutaneous wound repair is an intricate process whereby the skin reprograms itself after injury. In the mid-phase of wound repair, the proliferation, migration, and differentiation of cells are the major mechanisms to lead remodeling. We investigated the effect of BMM ((1E,2E)-1,2-bis((6-bromo-2H-chromen-3-yl)methylene)hydrazine), a novel synthetic material, on the migration and viability of keratinocytes or fibroblasts using the in vitro scratch woundhealing, electric cell-substrate imedance sensing (ECIS), invasion, and MTT assays. Cell migration-related factors were analyzed using western blot, and we found that treatment with BMM stimulated the EMT pathway and focal adhesion kinase (FAK)/Src signaling. Differentiation of HaCaT keratinocyte and fibroblast cells was also stimulated by BMM and specifically, NOX2/4 contributed to the activation of fibroblasts for wound healing. Furthermore, BMM treated HaCaT keratinocyte and fibroblast-co-cultured cells increased migration and differentiation. TGF-ß and Cyr61 were also secreted to a greater extent than in single cultured cells. In vivo experiments showed that treatment with BMM promotes wound closure by promoting re-epithelialization. In this study, we demonstrated that a novel synthetic material, BMM, is capable of promoting wound healing via the stimulation of re-epithelialization in the epidermis and the activation of fibroblasts in the dermis, in particular, via the acceleration of the interaction between the epidermis and dermis.


Subject(s)
Benzopyrans/pharmacology , Fibroblasts/drug effects , Hydrazines/pharmacology , Re-Epithelialization , Animals , Benzopyrans/chemistry , Cell Line , Cell Movement , Cell Proliferation , Cells, Cultured , Cysteine-Rich Protein 61/metabolism , Fibroblasts/metabolism , Fibroblasts/physiology , Focal Adhesion Protein-Tyrosine Kinases/metabolism , Humans , Hydrazines/chemistry , Keratinocytes/drug effects , Keratinocytes/metabolism , Keratinocytes/physiology , Male , Mice , Mice, Inbred ICR , NADPH Oxidases/metabolism , Signal Transduction , Transforming Growth Factor beta/metabolism , src-Family Kinases/metabolism
14.
Connect Tissue Res ; 59(3): 245-254, 2018 05.
Article in English | MEDLINE | ID: mdl-28750181

ABSTRACT

SIRT1 is a NAD-dependent protein deacetylase that participates in cellular regulation. The increased migration of fibroblasts is an important phenotype in fibroblast activation. The role of SIRT1 in cell migration remains controversial as to whether SIRT1 acts as an activator or suppressor of cell migration. Therefore, we have established the role of SIRT1 in the migration of human dermal fibroblasts and explored targets of SIRT1 during dermal fibroblast migration. SIRT1 and Cyr61 were expressed in human dermal fibroblasts and the stimulation with TGF-ß further induced their expression. Treatment with resveratrol (RSV), a SIRT1 agonist, or overexpression of SIRT1 also promoted the expression Cyr61 in human dermal fibroblasts, whereas the inhibition of SIRT1 activity by nicotinamide or knockdown of SIRT1 decreased the level of Cyr61, as well as TGF-ß or RSV-induced Cyr61 expression. Blocking of ERK signaling by PD98509 reduced the expression of Cyr61 induced by TGF-ß or RSV. TGF-ß, RSV, or SIRT1 overexpression enhanced ß-catenin as well as Cyr61 expression. This stimulation was reduced by the Wnt inhibitor XAV939. RSV increased migration and nicotinamide attenuated RSV-induced migration of human dermal fibroblasts. Furthermore, SIRT1 overexpression promoted cell migration, whereas blocking Cyr61 attenuated SIRT1-stimulated migration of human dermal fibroblasts. SIRT1 increased cell migration by stimulating Cyr61 expression and the ERK and Wnt/ß-catenin signaling. SIRT1-induced Cyr61 activity is very important for human dermal fibroblasts migration.


Subject(s)
Cysteine-Rich Protein 61/metabolism , Fibroblasts/metabolism , Sirtuin 1/metabolism , Transforming Growth Factor beta/metabolism , Cell Cycle/physiology , Cell Movement/physiology , Cells, Cultured , Child , Humans , Phenotype , Wnt Signaling Pathway/physiology , beta Catenin/metabolism
15.
Protein J ; 37(1): 82-92, 2018 02.
Article in English | MEDLINE | ID: mdl-29256161

ABSTRACT

A polyclonal antibody specific to an egg protein of Suminoe oyster Crassostrea ariakensis was previously developed in our laboratory to assess the reproductive life cycle of the oyster. The present study was undertaken to investigate vitellin of C. ariakensis (CAVt). Vitellin is an essential component of egg proteins in marine invertebrates as it provides energy and nutrients to the embryo and larvae. CAVt was purified from eggs of the oyster using ammonium sulfate precipitation followed by affinity chromatography with Concanavalin A-agarose. Native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate PAGE showed that CAVt is a high molecular weight [532 kiloDaltons (kDa)] protein, with multiple subunits. Similar to other vitellin proteins, it is a phospholipoglycoprotein composed of phospholipids (12.06%), carbohydrates (mannose, 10.08% or glucose, 9.84%), and alkali-labile phosphates (4.16%). Affinity chromatography, enzyme-linked immunosorbent aasay (ELISA) and western blot analysis revealed that CAVt is only present in the ovary, and two subunits of CAVt (72 and 35 kDa) are believed to be incorporated from the hemolymph into the oocyte. The antibody specific to CAVt (anti-CAVt), raised in rabbit, strongly cross reacted with the egg proteins of oyster species and scallops, suggesting that the antigenic epitopes are highly conserved among species. Our results suggest that the anti-CAVt antibody can be used to develop a tool similar to ELISA or western blotting for investigation of the effect of microorganisms on reproduction as well as the effect of chemicals on the endocrine system in C. ariakensis.


Subject(s)
Antibodies , Aquatic Organisms , Ostreidae , Ovum , Vitellins , Animals , Antibodies/chemistry , Antibodies/immunology , Aquatic Organisms/chemistry , Aquatic Organisms/immunology , Cross Reactions , Ostreidae/chemistry , Ostreidae/immunology , Ovum/chemistry , Ovum/immunology , Rabbits , Vitellins/chemistry , Vitellins/immunology , Vitellins/isolation & purification
16.
Exp Mol Med ; 49(3): e302, 2017 03 17.
Article in English | MEDLINE | ID: mdl-28303029

ABSTRACT

Keratinocyte-fibroblast interactions are critical for skin repair after injury. During the proliferative phase of wound healing, proliferation, migration and differentiation of these cells are the major mechanisms leading to tissue remodeling. We have previously reported that glycitin, a major soy isoflavone, stimulates dermal fibroblast proliferation; and the phytochemical, 4',6,7-trimethoxyisoflavone (TMF), induces migration of HaCaT keratinocyte cells. We therefore investigated whether these compounds display synergistic effects on skin cells during wound healing in vitro and in vivo. Co-treatment with TMF and glycitin synergistically promotes the proliferation and migration of both keratinocytes and dermal fibroblasts, with a 1:1 ratio of these compounds showing the greatest efficacy in our co-culture system. This keratinocyte-fibroblast interaction occurred via the secretion of TGF-ß, and the induction of differentiation and proliferation was confirmed in both indirect and direct co-culture assays. In an excisional and burn wound animal model, mice treated with a 1:1 ratio of TMF and glycitin showed faster wound closure, regeneration and scar reduction than even the positive control drug. These data indicate that two isoflavones, TMF and glycitin, act synergistically to promote wound healing and anti-scarring and could potentially be developed together as a bioactive therapeutic for wound treatment.


Subject(s)
Burns/drug therapy , Cicatrix/drug therapy , Fibroblasts/drug effects , Isoflavones/pharmacology , Keratinocytes/drug effects , Animals , Cell Movement , Cell Proliferation , Cells, Cultured , Female , Fibroblasts/metabolism , Humans , Isoflavones/therapeutic use , Keratinocytes/metabolism , Lymphotoxin-alpha/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Inbred ICR
17.
Int J Rheum Dis ; 20(10): 1499-1509, 2017 Oct.
Article in English | MEDLINE | ID: mdl-27456070

ABSTRACT

AIM: Peroxisome proliferator-activated receptor gamma (PPARγ) agonists have anti-inflammatory properties that reduce inflammatory cytokine production in rheumatoid arthritis (RA). Cysteine-rich angiogenic inducer 61 (Cyr61) is associated with diseases related to chronic inflammation. The aim of this study was to investigate the mechanisms underlying the effects of PPARγ agonists on tumor necrosis factor (TNF)-α-induced fibroblast-like synoviocyte (FLS) invasion and migration, as well as Cyr61 production, in RA-FLS. METHODS: FLS were cultured with TNF-α and Cyr61 in the presence or absence of PPARγ agonists. Matrix metalloproteinase and Cyr61 expression levels in RA-FLS and culture supernatants were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. The migration and invasion phenotypes of RA-FLS were determined by wound healing and Boyden chamber assays. RESULTS: Cyr61 protein was expressed in RA-FLS, and its intracellular expression and secretion levels were increased by TNF-α. Moreover, Cyr61 directly promoted RA-FLS migration and invasion. Rosiglitazone (RSG) significantly decreased TNF-α-induced Cyr61 expression. RSG decreased TNF-α-induced nuclear factor (NF)-κB activation and inhibitor of κBα degradation. Furthermore, RSG inhibited TNF-α-induced RA-FLS migration and invasion and decreased Cyr61 treatment-induced RA-FLS invasion. Finally, blocking Cyr61 significantly attenuated TNF-α-induced migration. CONCLUSIONS: Our results demonstrate for the first time that PPARγ agonists may have beneficial effects on the migration and invasion of RA-FLS via the downregulation of Cyr61. Therefore, PPARγ agonists could be potential treatment targets for RA.


Subject(s)
Antirheumatic Agents/pharmacology , Arthritis, Rheumatoid/drug therapy , Cell Movement/drug effects , Cysteine-Rich Protein 61/metabolism , Fibroblasts/drug effects , PPAR gamma/agonists , Synovial Membrane/drug effects , Synoviocytes/drug effects , Thiazolidinediones/pharmacology , Arthritis, Rheumatoid/genetics , Arthritis, Rheumatoid/metabolism , Arthritis, Rheumatoid/pathology , Cells, Cultured , Cysteine-Rich Protein 61/genetics , Dose-Response Relationship, Drug , Down-Regulation , Fibroblasts/metabolism , Fibroblasts/pathology , Humans , NF-KappaB Inhibitor alpha/metabolism , NF-kappa B/metabolism , PPAR gamma/metabolism , Phenotype , Rosiglitazone , Signal Transduction/drug effects , Synovial Membrane/metabolism , Synovial Membrane/pathology , Synoviocytes/metabolism , Synoviocytes/pathology , Tumor Necrosis Factor-alpha/pharmacology
18.
EXCLI J ; 15: 329-42, 2016.
Article in English | MEDLINE | ID: mdl-27366143

ABSTRACT

The present study was designed to investigate the anti-cancer effects of Sea hare eggs (SE) in U937 cells and its major active components. The aqueous extract of SE (ASE), which contained the highest protein content, dose-dependently inhibited the cancer cell's growth (IC50 value, 10.42 ± 0.5 µg/mL). Additionally, ASE markedly caused DNA damage by inducing apoptotic body formation, DNA fragmentation, and accumulation of sub-G1 DNA contents. ASE induced apoptosis by activating caspase-3 and 9 and poly (ADP-ribose) polymerase (PARP) by regulating the expression of Bcl-2/Bax. Moreover, among its molecular weight fractions, the > 30 kDa fraction showed the highest cell-growth-inhibitory effects, which was inhibited by heat treatment. Furthermore, the > 30 kDa fraction had markedly higher glycine content than the ASE. The presence of two protein bands at around 16 and 32 kDa was identified. In addition, two fractions, F1 and F2, were obtained using anion-exchange chromatography, with the F1 having an improved cell-growth-inhibitory effect than the > 30 kDa fraction. Taken together, these results suggest that the ASE contains glycine-rich proteins, including the active 16 and 32 kDa proteins, which account for its anti-cancer effects by inducing apoptosis via regulation of the mitochondrial pathway.

19.
Biochem Biophys Res Commun ; 472(2): 332-8, 2016 Apr 01.
Article in English | MEDLINE | ID: mdl-26923073

ABSTRACT

Flavonoids are plant secondary compounds with various pharmacological properties. We previously showed that one flavonoid, trimethoxyisoflavone (TMF), could promote wound healing by inducing keratinocyte migration. Here, we screened TMF derivatives for enhanced activity and identified one compound, 2',6 Dichloro-7-methoxyisoflavone (DCMF), as most effective at promoting migration in a scratch wound assay. Using the HaCaT keratinocyte cell line, we found DCMF treatment induced phosphorylation of both FAK and Src, and increased keratinocyte migration. DCMF-induced Src kinase could promote activation of ERK, AKT, and p38 signaling pathways, and DCMF-induced secretion of matrix metalloproteinase (MMP)-2 and MMP-9 and partial epithelial-mesenchymal transition (EMT), whereas Src inhibition abolished DCMF-induced EMT. Using an in vivo excisional wound model, we observed improved wound closure and re-epithelialization in DCMF-treated mice, as compared to controls. Collectively, our data demonstrate that DCMF induces cell migration and promotes wound healing through activation of Src/FAK, ERK, AKT, and p38 MAPK signaling.


Subject(s)
Cell Movement/physiology , Focal Adhesion Kinase 1/metabolism , Isoflavones/administration & dosage , Keratinocytes/physiology , Wound Healing/drug effects , src-Family Kinases/metabolism , Animals , Cell Line , Cell Movement/drug effects , Dose-Response Relationship, Drug , Enzyme Activation/drug effects , Humans , Isoflavones/chemical synthesis , Keratinocytes/drug effects , Male , Mice , Mice, Inbred ICR , Signal Transduction/drug effects , Signal Transduction/physiology , Wound Healing/physiology
20.
Cancer Res Treat ; 48(3): 1141-53, 2016 Jul.
Article in English | MEDLINE | ID: mdl-26693911

ABSTRACT

PURPOSE: One of the features in cancer development is the migration of cancer cells to form metastatic lesions. CYR61 protein promotes migration and the epithelial-mesenchymal transition in several cancer cell types. Evidence suggests that CYR61 and dexamethasone are relevant to colorectal cancer. However, relationships between them and colorectal cancer are still unclear. Understanding the molecular mechanism of colorectal cancer progression related with CYR61 and dexamethasone, which is widely used for combination chemotherapy, is necessary for improved therapy. MATERIALS AND METHODS: We used colorectal cancer cells, HCT116, co-treated with transforming growth factor ß1 (TGF-ß1) and dexamethasone to examine the inhibitory migration effect of dexamethasone by migratory assay. Alternatively, both migratory pathways, expression of AKT and ERK, and the target factor CYR61 was also tested by co-treatment with TGF-ß1 and dexamethasone. RESULTS: We report that dexamethasone significantly inhibited TGF-ß1-induced cell migration, without affecting cell proliferation. Importantly, we observed that TGF-ß1 promoted the epithelial-mesenchymal transition process and that dexamethasone co-treatment abolished this effect. ERK and AKT signaling pathways were found to mediate TGF-ß1-induced migration, which was inhibited by dexamethasone. In addition, TGF-ß1 treatment induced CYR61 expression whereas dexamethasone reduced it. These observations were compatible with the modulation of migration observed following treatment of HCT116 cells with human recombinant CYR61 and anti-CYR61 antibody. Our results also indicated that TGF-ß1 enhanced collagen I and reduced matrix metalloproteinase 1 expression, which was reversed by dexamethasone treatment. CONCLUSION: These findings suggested that dexamethasone inhibits AKT and ERK phosphorylation, leading to decreased CYR61 expression, which in turn blocks TGF-ß1-induced migration.


Subject(s)
Antineoplastic Agents, Hormonal/pharmacology , Cell Movement/drug effects , Colorectal Neoplasms/pathology , Cysteine-Rich Protein 61/metabolism , Dexamethasone/pharmacology , MAP Kinase Signaling System/drug effects , Transforming Growth Factor beta1/metabolism , Antibodies/pharmacology , Cadherins , Cell Proliferation/drug effects , Collagen Type I/metabolism , Cysteine-Rich Protein 61/antagonists & inhibitors , Disease Progression , Down-Regulation , Epithelial-Mesenchymal Transition/drug effects , Extracellular Signal-Regulated MAP Kinases/metabolism , HCT116 Cells , Humans , Matrix Metalloproteinase 1/metabolism , Phosphorylation , Proto-Oncogene Proteins c-akt/metabolism , Recombinant Proteins/metabolism , Up-Regulation
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