ABSTRACT
Lipoic acid (alpha lipoic acid, thioctic acid) is a popular over-the-counter antioxidant and insulin-mimetic supplement under investigation in a variety of conditions including multiple sclerosis, diabetes, and schizophrenia. Unfortunately, high-grade proteinuria was an unexpected adverse event specific to the treatment arm of our clinical trial investigating lipoic acid supplementation in patients with multiple sclerosis. This observation led to detection of similar patients in our nephrology practice. Here, we describe four biopsy-proven cases of neural epidermal growth factor-like 1 (NELL1)-associated membranous nephropathy following lipoic acid supplementation and a fifth suspected case. Discontinuation of lipoic acid and supportive therapy resulted in remission.
Subject(s)
Glomerulonephritis, Membranous , Thioctic Acid , Calcium-Binding Proteins , Dietary Supplements , EGF Family of Proteins , Glomerulonephritis, Membranous/diagnosis , Glomerulonephritis, Membranous/drug therapy , Humans , Proteinuria/chemically induced , Proteinuria/drug therapy , Thioctic Acid/adverse effectsABSTRACT
<p><b>OBJECTIVE</b>To investigate effects of retinol on the expressions of epidermal growth factor (EGF), stem cell factor (SCF), colony-stimulating factor 1 (CSF1) and leukemia inhibitory factor (LIF) in cultured human umbilical-derived mesenchymal stem cells (UCMSCs).</p><p><b>METHODS</b>Human UCMSCs were isolated from human umbilical cord and identified for immunophenotypes. The cells were then cultured in DMEM/F12 media supplemented with 12% fetal bovine serum (FBS), 12% FBS+1 µmol/L retinol, 15% knockout serum replacement (KSR) and 15% KSR+ 1 µmol/L retinol. The expressions of the cytokines EGF, SCF, CSF1 and LIF in the cells were detected using RT-PCR and ELISA.</p><p><b>RESULTS</b>The isolated cells exhibited characteristic immunophenotypes of human UCMSCs and expressed EGF, CSF1 and SCF at both mRNA and protein levels but not LIF protein. Retinol (1 µmol/L) significantly promoted the expressions of SCF and CSF1 at both mRNA and protein levels but did not result in changes of EGF and LIF expressions in human UCMSCs.</p><p><b>CONCLUSION</b>Retinol at the concentration of 1 µmol/L can promote expression of SCF and CSF1 in human UCMSCs in vitro.</p>
Subject(s)
Humans , Cell Differentiation , Cells, Cultured , EGF Family of Proteins , Metabolism , Immunophenotyping , Leukemia Inhibitory Factor , Metabolism , Macrophage Colony-Stimulating Factor , Metabolism , Mesenchymal Stem Cells , Metabolism , Stem Cell Factor , Metabolism , Umbilical Cord , Cell Biology , Vitamin A , PharmacologyABSTRACT
OBJECTIVE: To investigate effects of retinol on the expressions of epidermal growth factor (EGF), stem cell factor (SCF), colony-stimulating factor 1 (CSF1) and leukemia inhibitory factor (LIF) in cultured human umbilical-derived mesenchymal stem cells (UCMSCs). METHODS: Human UCMSCs were isolated from human umbilical cord and identified for immunophenotypes. The cells were then cultured in DMEM/F12 media supplemented with 12% fetal bovine serum (FBS), 12% FBS+1 µmol/L retinol, 15% knockout serum replacement (KSR) and 15% KSR+ 1 µmol/L retinol. The expressions of the cytokines EGF, SCF, CSF1 and LIF in the cells were detected using RT-PCR and ELISA. RESULTS: The isolated cells exhibited characteristic immunophenotypes of human UCMSCs and expressed EGF, CSF1 and SCF at both mRNA and protein levels but not LIF protein. Retinol (1 µmol/L) significantly promoted the expressions of SCF and CSF1 at both mRNA and protein levels but did not result in changes of EGF and LIF expressions in human UCMSCs. CONCLUSION: Retinol at the concentration of 1 µmol/L can promote expression of SCF and CSF1 in human UCMSCs in vitro.
Subject(s)
EGF Family of Proteins/metabolism , Leukemia Inhibitory Factor/metabolism , Macrophage Colony-Stimulating Factor/metabolism , Mesenchymal Stem Cells/drug effects , Stem Cell Factor/metabolism , Vitamin A/pharmacology , Cell Differentiation , Cells, Cultured , Humans , Immunophenotyping , Mesenchymal Stem Cells/metabolism , Umbilical Cord/cytologyABSTRACT
INTRODUCTION: Epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) play an important role in extracellular matrix mineralization, a complex process required for proper bone regeneration, one of the biggest challenges in dentistry. The purpose of this study was to evaluate the osteogenic potential of EGF and bFGF on dental pulp stem cells (DPSCs). MATERIAL AND METHODS: Human DPSCs were isolated using CD105 magnetic microbeads and characterized by flow cytometry. To induce osteoblast differentiation, the cells were cultured in osteogenic medium supplemented with EGF or bFGF at a low concentration. Cell morphology and expression of CD146 and CD10 surface markers were analyzed using fluorescence microscopy. To measure mineralization, an alizarin red S assay was performed and typical markers of osteoblastic phenotype were evaluated by RT-PCR. RESULTS: EGF treatment induced morphological changes and suppression of CD146 and CD10 markers. Additionally, the cells were capable of producing calcium deposits and increasing the mRNA expression to alkaline phosphatase (ALP) and osteocalcin (OCN) in relation to control groups (p < 0.001). However, bFGF treatment showed an inhibitory effect. CONCLUSION: These data suggests that DPSCs in combination with EGF could be an effective stem cell-based therapy for bone tissue engineering applications in periodontics and oral implantology.
Subject(s)
Cell Differentiation/drug effects , Dental Pulp/cytology , EGF Family of Proteins/pharmacology , Fibroblast Growth Factor 2/pharmacology , Osteogenesis/drug effects , Stem Cells/drug effects , CD146 Antigen/metabolism , Cells, Cultured , Flow Cytometry , Humans , Immunohistochemistry , Neprilysin/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Stem Cells/metabolismABSTRACT
BACKGROUND: Ulcerative colitis is a chronic inflammatory disease and involves multiple etiological factors. Acetic acid (AA)-induced colitis is a reproducible and simple model, sharing many characteristics with human colitis. N-acetylcysteine (NAC) has been widely used as an antioxidant in vivo and in vitro. NAC can affect several signaling pathways involving in apoptosis, angiogenesis, cell growth and arrest, redox-regulated gene expression, and inflammatory response. Therefore, NAC may not only protect against the direct injurious effects of oxidants, but also beneficially alter inflammatory events in colitis. This study was conducted to investigate whether NAC could alleviate the AA-induced colitis in a porcine model. METHODS: Weaned piglets were used to investigate the effects of NAC on AA-induced colitis. Severity of colitis was evaluated by colon histomorphology measurements, histopathology scores, tissue myeloperoxidase activity, as well as concentrations of malondialdehyde and pro-inflammatory mediators in the plasma and colon. The protective role of NAC was assessed by measurements of antioxidant status, growth modulator, cell apoptosis, and tight junction proteins. Abundances of caspase-3 and claudin-1 proteins in colonic mucosae were determined by the Western blot method. Epidermal growth factor receptor, amphiregulin, tumor necrosis factor-alpha (TNF-α), and toll-like receptor 4 (TLR4) mRNA levels in colonic mucosae were quantified using the real-time fluorescent quantitative PCR. RESULTS: Compared with the control group, AA treatment increased (P < 0.05) the histopathology scores, intraepithelial lymphocyte (IEL) numbers and density in the colon, myeloperoxidase activity, the concentrations of malondialdehyde and pro-inflammatory mediators in the plasma and colon, while reducing (P < 0.05) goblet cell numbers and the protein/DNA ratio in the colonic mucosa. These adverse effects of AA were partially ameliorated (P < 0.05) by dietary supplementation with NAC. In addition, NAC prevented the AA-induced increase in caspase-3 protein, while stimulating claudin-1 protein expression in the colonic mucosa. Moreover, NAC enhanced mRNA levels for epidermal growth factor and amphiregulin in the colonic mucosa. CONCLUSION: Dietary supplementation with NAC can alleviate AA-induced colitis in a porcine model through regulating anti-oxidative responses, cell apoptosis, and EGF gene expression.
Subject(s)
Acetic Acid , Acetylcysteine/pharmacology , Colitis, Ulcerative , Colitis/prevention & control , Free Radical Scavengers/pharmacology , Acetylcysteine/therapeutic use , Amphiregulin , Animals , Apoptosis/drug effects , Caspase 3/drug effects , Caspase 3/metabolism , Claudin-1/drug effects , Claudin-1/metabolism , Colitis/chemically induced , Colitis/pathology , Colon/drug effects , Colon/metabolism , Colon/pathology , Dietary Supplements , Dinoprostone/metabolism , Disease Models, Animal , EGF Family of Proteins , Epidermal Growth Factor/blood , Epidermal Growth Factor/drug effects , ErbB Receptors/drug effects , ErbB Receptors/genetics , Free Radical Scavengers/therapeutic use , Glycoproteins/drug effects , Glycoproteins/genetics , Intercellular Signaling Peptides and Proteins/genetics , Interleukin-6/metabolism , Intestinal Mucosa/drug effects , Intestinal Mucosa/metabolism , Intestinal Mucosa/pathology , Swine , Toll-Like Receptor 4/drug effects , Toll-Like Receptor 4/genetics , Transforming Growth Factor alpha/drug effects , Transforming Growth Factor alpha/metabolism , Tumor Necrosis Factor-alpha/drug effects , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Decreased bone mineral density (BMD) represents an extraintestinal complication of inflammatory bowel disease (IBD). Vitamin D3 has been considered a viable adjunctive therapy in IBD. However, vitamin D3 plays a pleiotropic role in bone modeling and regulates the bone formation-resorption balance, depending on the physiological environment, and supplementation during active IBD may have unintended consequences. We evaluated the effects of vitamin D3 supplementation during the active phase of disease on colonic inflammation, BMD, and bone metabolism in an adoptive IL-10-/- CD4⺠T cell transfer model of chronic colitis. High-dose vitamin D3 supplementation for 12 days during established disease had negligible effects on mucosal inflammation. Plasma vitamin D3 metabolites correlated with diet, but not disease, status. Colitis significantly reduced BMD. High-dose vitamin D3 supplementation did not affect cortical bone but led to a further deterioration of trabecular bone morphology. In mice fed a high vitamin D3 diet, colitis more severely impacted bone formation markers (osteocalcin and bone alkaline phosphatase) and increased bone resorption markers, ratio of receptor activator of NF-κB ligand to osteoprotegrin transcript, plasma osteoprotegrin level, and the osteoclast activation marker tartrate-resistant acid phosphatase (ACp5). Bone vitamin D receptor expression was increased in mice with chronic colitis, especially in the high vitamin D3 group. Our data suggest that vitamin D3, at a dose that does not improve inflammation, has no beneficial effects on bone metabolism and density during active colitis or may adversely affect BMD and bone turnover. These observations should be taken into consideration in the planning of further clinical studies with high-dose vitamin D3 supplementation in patients with active IBD.
Subject(s)
Bone and Bones/drug effects , Bone and Bones/metabolism , Cholecalciferol/pharmacology , Colitis/complications , Vitamins/pharmacology , Adoptive Transfer , Amphiregulin , Animal Feed , Animals , Bone Density/drug effects , CD4-Positive T-Lymphocytes/physiology , Cholecalciferol/administration & dosage , Chronic Disease , Colitis/metabolism , Diet , EGF Family of Proteins , Gene Deletion , Glycoproteins/metabolism , Intercellular Signaling Peptides and Proteins/metabolism , Interleukin-10/genetics , Interleukin-10/metabolism , Mice , Mice, Knockout , Vitamins/administration & dosageABSTRACT
BACKGROUND: Colostrum contains a wide variety of crucial nutritional elements including growth factors for newborn infants to adapt to the extrauterine environment. OBJECTIVE: To investigate the clinical significance of epidermal growth factor receptor ligands in milk during the first month of lactation. METHODS: The concentrations of epidermal growth factor (EGF), amphiregulin (AR) and transforming growth factor-α (TGF-α) in milk sampled from a total of 31 normal mothers at days 1-3, 5, and 30 postpartum were examined using ELISA. RESULTS: At days 1-3, the concentration of EGF was extremely high [131.6 ± 20.4 (mean ± SEM) ng/ml] compared to that of AR (4,197.2 ± 1,055.2 pg/ml) or TGF-α (261.7 ± 33.6 pg/ml), while the concentration of AR was significantly elevated compared to that of TGF-α. At days 5 and 30, the concentration of EGF was significantly elevated compared to that of AR or TGF-α. In 16 mothers among the same 31 subjects, samples were longitudinally obtained on days 1, 2, 5, and 30 postpartum. Concentrations of AR were higher on days 1 and 2 and rapidly declined to below 1 ng/ml on day 5, and were maintained at lower levels on day 30. Concentrations of EGF were high on day 1 (greater than 10 ng/ml) but gradually declined by days 2, 5, and 30. Concentrations of TGF-α remained at lower levels of below 1 ng/ml throughout the lactation period from days 1 to 30. CONCLUSION: These results suggested that EGF and amphiregulin in colostrum might contribute to the early stage of development of neonatal gastrointestinal function.
Subject(s)
Colostrum/metabolism , Epidermal Growth Factor/metabolism , Glycoproteins/metabolism , Intercellular Signaling Peptides and Proteins/metabolism , Milk, Human/metabolism , Transforming Growth Factor alpha/metabolism , Adult , Amphiregulin , EGF Family of Proteins , Female , Humans , Lactation , Young AdultABSTRACT
Mycoplasma pneumoniae (M. pneumoniae) is increasingly recognized as a major cause of acute respiratory tract infections. Today, macrolides are used in the primary treatment of M. pneumoniae infection. However, with the increasing prevalence of strains resistant to macrolides, as well as reports of toxicity and adverse side effects, it is necessary to develop an alternative therapeutic agent. A compound recipe - Qinbaiqingfei pellets (Qinbai) - have already been approved in China as the first effective traditional Chinese medicine to be used against M. pneumoniae. Herein, we characterize the mechanism by which Qinbai interacts with M. pneumoniae and lung epithelial cells. The fact that Baicalin is the key component of Qingbai leads us to believe its study is important to elucidating the mechanism of the action of Qinbai. In this study, we describe the complex impact of Baicalin on the adhesin protein P1 of M. pneumoniae and on the expression of epidermal growth factor (EGF) in BALB/c mice and A549 cells infected with M. pneumonia. We draw the conclusion that Baicalin not only cured M. pneumoniae infection by inhibiting P1 expression, but also enhanced the repair of lung epithelial cells by upregulating EGF. Finally, we demonstrate that Baicalin plays a role in Qinbai treatment.
Subject(s)
Adhesins, Bacterial/genetics , EGF Family of Proteins/genetics , Flavonoids/pharmacology , Gene Expression Regulation/drug effects , Adhesins, Bacterial/metabolism , Animals , EGF Family of Proteins/metabolism , Fluorescent Antibody Technique , Humans , Lung/drug effects , Lung/pathology , Mice, Inbred BALB C , Mycoplasma pneumoniae/drug effects , Mycoplasma pneumoniae/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
We report the cloning and characterization of a novel epidermal growth factor (EGF) domain gene that was identified in a retroviral gene entrapment screen and is expressed in endothelial cells. This gene encodes a protein of 278 amino acids with an amino-terminal signal peptide and two centrally located EGF-like domains. We have named this novel gene in accordance with the guidelines of the Mouse Genome Informatics group Egfl7, for EGF-like domain 7. Egfl7 mRNA is expressed in highly vascularized adult tissues such as the lung, heart, uterus, and ovary. In addition, Egfl7 is expressed early during mouse embryogenesis and in undifferentiated murine embryonic stem cells. The analysis of Egfl7 expression in embryonic day 9.5 embryos by in situ hybridization indicates that Egfl7 is expressed in vascular structures in both the embryo proper and the yolk sac and at sites of mesodermal precursors of angioblasts. Within the cell, EGFL7 protein is localized to the endoplasmic reticulum and Golgi apparatus, suggesting that the protein is targeted for secretion. Indeed, recombinant EGFL7 is readily detectable in the supernatant media of transiently transfected HEK293 cells. We also report the identification of an Egfl7 paralog, Egfl8, and show that EGFL8 protein shares similar domains and molecular weight with EGFL7.
Subject(s)
Endothelial Cells/metabolism , Gene Expression Regulation , Proteins/genetics , Aging/physiology , Amino Acid Sequence , Animals , Calcium-Binding Proteins , Cell Line , Cloning, Molecular , DNA, Complementary/genetics , EGF Family of Proteins , Embryo, Mammalian/blood supply , Embryo, Mammalian/cytology , Embryo, Mammalian/embryology , Embryo, Mammalian/metabolism , Embryonic Development , Endoplasmic Reticulum/metabolism , Endothelial Cells/cytology , Gene Expression Profiling , Golgi Apparatus/metabolism , Humans , Mesoderm/metabolism , Mice , Molecular Sequence Data , Proteins/chemistry , Proteins/metabolism , Sequence Alignment , Yolk Sac/metabolismABSTRACT
The recruitment and proliferation of smooth muscle cells and pericytes are two key events for the stabilization of newly formed capillaries during angiogenesis and, when out of control in the adult, are the main causes of arteriosclerosis. We have identified a novel gene, named VE-statin for vascular endothelial-statin, which is expressed specifically by endothelial cells of the developing mouse embryo and in the adult, and in early endothelial progenitors. The mouse and human VE-statin genes have been located on chromosome 2 and 9, respectively, they span >10 kbp and are transcribed in two major variants arising from independent initiation sites. The VE-statin transcripts code for a unique protein of 30 kDa that contains a signal peptide and two epidermal growth factor (EGF)-like modules. VE-statin is found in the cellular endoplasmic reticulum and secreted in the cell supernatant. Secreted VE-statin inhibits platelet-derived growth factor (PDGF)-BB-induced smooth muscle cell migration, but has no effects on endothelial cell migration. VE-statin is the first identified inhibitor of mural cell migration specifically produced by endothelial cells.
Subject(s)
Endothelial Growth Factors/physiology , Endothelium, Vascular/physiology , Growth Inhibitors/physiology , Muscle, Smooth, Vascular/cytology , Proteins/physiology , Transcription Factors/physiology , Amino Acid Sequence , Animals , Base Sequence , Calcium-Binding Proteins , Cell Division , Cell Line , Cell Movement , Cells, Cultured , Chromosomes, Human, Pair 9/genetics , Cloning, Molecular , DNA, Complementary/genetics , DNA-Binding Proteins , EGF Family of Proteins , Endothelial Growth Factors/genetics , Endothelium, Vascular/growth & development , Growth Inhibitors/genetics , Humans , In Situ Hybridization, Fluorescence , In Vitro Techniques , Kruppel-Like Transcription Factors , Mice , Molecular Sequence Data , Muscle, Smooth, Vascular/growth & development , Muscle, Smooth, Vascular/physiology , Neovascularization, Physiologic , Proteins/genetics , Transcription Factors/geneticsABSTRACT
BACKGROUND & AIMS: Epidermal growth factor (EGF)-like growth factors are induced after acute gastric injury and may play an important role in mucosal repair. However, the mechanisms that trigger these growth factors are poorly understood. We determined the role of EGF receptor (EGFR) in stress-induced expression of heparin-binding EGF-like growth factor (HB-EGF) in a rat gastric epithelial cell line (RGM1 cells). METHODS: RGM1 cells were transfected with a plasmid containing complementary DNA encoding a dominant-negative human EGFR (HERCD533). Cells were treated with hydrogen peroxide (0-400 micromol/L) or sorbitol (600 mmol/L). Tyrosine phosphorylation of EGFR was determined by immunoprecipitation and Western blotting with an antiphosphotyrosine antibody. HB-EGF messenger RNA and protein were determined with Northern and Western blotting, respectively. Cell growth was evaluated by cell number and [(3)H]thymidine incorporation. RESULTS: Oxidative stress and osmotic stress induced tyrosine phosphorylation of EGFR within 2 minutes, followed by a marked increase in HB-EGF and amphiregulin transcripts in RGM1 cells. Introduction of HERCD533 into the cells inhibited not only tyrosine phosphorylation of EGFR but also growth response to EGF. Furthermore, oxidative stress-induced HB-EGF messenger RNA expression was impaired in HERCD533-expressing cells. CONCLUSIONS: EGFR plays a crucial role in the stress-induced expression of EGF-like growth factors in gastrointestinal epithelial cells.
Subject(s)
Epidermal Growth Factor/genetics , ErbB Receptors/genetics , ErbB Receptors/metabolism , Gastric Mucosa/metabolism , Intercellular Signaling Peptides and Proteins , Oxidative Stress/physiology , Amphiregulin , Animals , Blotting, Western , Culture Media, Serum-Free/pharmacology , DNA, Complementary , EGF Family of Proteins , Epidermal Growth Factor/analysis , Epidermal Growth Factor/metabolism , Epidermal Growth Factor/pharmacology , Gastric Mucosa/cytology , Gastric Mucosa/drug effects , Gene Expression Regulation/drug effects , Gene Expression Regulation/physiology , Glycoproteins/genetics , Growth Substances/genetics , Heparin-binding EGF-like Growth Factor , Ligands , Phosphorylation , Promoter Regions, Genetic/physiology , Proto-Oncogene Proteins c-met/metabolism , RNA, Messenger/metabolism , Rats , Rats, Wistar , Receptor, ErbB-2/metabolism , Receptor, ErbB-3/metabolism , Transfection , Transforming Growth Factor alpha/genetics , Tyrosine/metabolismABSTRACT
Amphiregulin and transforming growth factor-alpha, agonists for the epidermal growth factor receptor, are the major autocrine growth factors for cultured keratinocytes, and their substantial overexpression in psoriatic lesions suggests that they are crucial to the basal hyperplasia that characterizes psoriasis. Amphiregulin binds to heparin and related highly sulfated polysaccharides, and exogenous heparin blocks its growth factor activity, rationalizing previous reports that psoriasis responds to heparin therapy. Differentiating keratinocytes produce increased amounts of protein-bound as well as free-chain heparan sulfates, which may function physiologically as amphiregulin antagonists. By promoting keratinocyte synthesis of these heparan sulfates, glucosamine administration may inhibit amphiregulin function and thus provide therapeutic benefit in psoriasis. Concurrent ingestion of fish oil, by impeding the excessive activation of protein kinase C, may decrease keratinocyte production of amphiregulin and other autocrine growth factors, thus complementing the postulated benefits of glucosamine.