Pomegranate Bioactive Constituents Suppress Cell Proliferation and Induce Apoptosis in an Experimental Model of Hepatocellular Carcinoma: Role of Wnt/β-Catenin Signaling Pathway.
Immunohistochemistry; Gene Expression; Apoptosis; Rats; Pomegranate; Post Hoc Analysis; One-Way Analysis of Variance; Structural Equation Modeling; Experimental Studies; Protocols; Carcinoma; Blotting; Western; Hepatocellular; Animal Studies; In Vivo Studies
Bhatia Deepak; Thoppil Roslin J; Mandal Animesh; Samtani Karishma A; Darvesh Altaf S; Bishayee Anupam
Evidence-based Complementary & Alternative Medicine (eCAM)
2013
2013-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1155/2013/371813" target="_blank" rel="noreferrer noopener">10.1155/2013/371813</a>
Apocynum Tablet Protects against Cardiac Hypertrophy via Inhibiting AKT and ERK1/2 Phosphorylation after Pressure Overload.
Mice; Gene Expression Profiling; Signal Transduction; Echocardiography; Staining and Labeling; Descriptive Statistics; Funding Source; Post Hoc Analysis; One-Way Analysis of Variance; T-Tests; Blotting; Western; Animal Studies; In Vivo Studies; Vasoconstriction; Phosphorylation – Drug Effects; Aorta – Physiopathology; Heart Diseases – Prevention and Control; Hypertrophy – Prevention and Control; Plant Extracts – Therapeutic Use; Protein Kinases – Antagonists and Inhibitors
Qi Jianyong; Liu Qin; Gong Kaizheng; Yu Juan; Wang Lei; Guo Liheng; Zhou Miao; Wu Jiashin; Zhang Minzhou
Evidence-based Complementary & Alternative Medicine (eCAM)
2014
2014-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1155/2014/769515" target="_blank" rel="noreferrer noopener">10.1155/2014/769515</a>
The expression of a catalytically active cholesterol 7 alpha-hydroxylase cytochrome P450 in Escherichia coli.
Gene Expression; Amino Acid Sequence; Base Sequence; Polymerase Chain Reaction; Liver/enzymology; Catalysis; Molecular Sequence Data; Substrate Specificity; Cholesterol/metabolism; Cholesterol 7-alpha-Hydroxylase/*genetics; DNA/genetics; Codon; Escherichia coli/*enzymology; Plasmids; Chromatography; Blotting; Western; Electrophoresis; Polyacrylamide Gel; Liquid; Microsomes
We have recently cloned a full-length cDNA encoding the rat hepatic cholesterol 7 alpha-hydroxylase cytochrome P450 (P450c7) (Li, Y. C., Wang, D. P., and Chiang, J. Y. L. (1990) J. Biol. Chem. 265, 12012-12019), which catalyzes the rate-limiting reaction of bile acid synthesis in the liver. By using the polymerase chain reaction, we have designed two P450c7 cDNAs. One has the second Met codon deleted and the third Thr codon replaced with an Ala. The other lacks codons for the NH2-terminal hydrophobic sequence of amino acids 2-24 (P450c7 delta 2-24). The cDNAs were separately cloned into the expression vector pKK233-2 and transformed into Escherichia coli. After induction with isopropyl-beta-D-thiogalactopyranoside, bacteria harboring recombinant plasmids expressed a polypeptide which reacted with the antibody against cholesterol 7 alpha-hydroxylase in immunoblots. The slightly modified full-length enzyme was expressed to 0.2% of the total bacterial lysate and was located in the membrane fraction, whereas P450c7 delta 2-24 was expressed at a 10-fold higher level (2%), of which 85% was in the cytosol and the remaining associated with the membranes. We have purified P450c7 delta 2-24 which showed a typical reduced-CO difference spectrum of cytochrome P450 and reconstituted cholesterol 7 alpha-hydroxylase activity in the presence of NADPH-cytochrome P450 reductase. P450c7 delta 2-24 has a similar Km for cholesterol (24.6 microM) but a lower Vmax (0.10 nmol/min) and a lower turnover number (1.93 min-1) as compared with the enzyme isolated from rat liver microsomes. The purified P450c7 delta 2-24 has an unique hydrophilic NH2 terminus and contains monomers and dimers in equal amounts. This is the first report demonstrating that a genetically engineered cytochrome P450 enzyme lacking a typical NH2-terminal hydrophobic sequence is mainly cytosolic and catalytically active.
Li Y C; Chiang J Y
The Journal of biological chemistry
1991
1991-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
Black currant anthocyanins abrogate oxidative stress through Nrf2- mediated antioxidant mechanisms in a rat model of hepatocellular carcinoma.
Male; Animals; Rats; Diet; Signal Transduction; NF-E2-Related Factor 2/genetics/*metabolism; *Phytotherapy; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Alkylating Agents/toxicity; Anthocyanins/*therapeutic use; Antioxidants/therapeutic use; Diethylnitrosamine/toxicity; Glutathione Transferase; Immunoenzyme Techniques; Lipid Peroxidation/drug effects; Nitric Oxide Synthase Type II/genetics/metabolism; Oxidative Stress/*drug effects; Ribes/*chemistry; Tyrosine/analogs & derivatives/metabolism; Carcinoma; Sprague-Dawley; Blotting; Western; RNA; Liver Neoplasms; Messenger/genetics; Experimental/chemically induced/metabolism/*prevention & control; Hepatocellular/chemically induced/metabolism/*prevention & control
Hepatocellular carcinoma (HCC), considered to be one of the most lethal cancers with almost \textgreater 1 million deaths reported annually worldwide, remains a devastating disease with no known effective cure. Hence, chemopreventive strategies come into play, offering an effective and safe mode of treatment, ideal to ward off potential cancer risks and mortality. A major predisposing condition, pertinent to the development and progression of HCC is oxidative stress. We previously reported a striking chemopreventive effect of anthocyanin-rich black currant skin extract (BCSE) against diethylnitrosamine (DENA)-initiated hepatocarcinogenesis in rats. The current study aims to elucidate the underlying antioxidant mechanisms of black currant anthocyanins implicated in the previously observed chemopreventive effects against experimental hepatocarcinogenesis. Dietary BCSE (100 and 500 mg/kg) administered four weeks before and 18 weeks after DENA challenge decreased abnormal lipid peroxidation, protein oxidation, and expression of inducible nitric oxide synthase (iNOS) and 3-nitrotyrosine (3-NT) in a dose-responsive fashion. Mechanistic studies revealed that BCSE upregulated the gene expression of a number of hepatic antioxidant and carcinogen detoxifying enzymes, such as NAD(P)H:quinone oxidoreductase, glutathione S-transferase, and uridine diphosphate-glucuronosyltransferase isoenzymes, in DENA-initiated animals. Protein and mRNA expressions of nuclear factor E2-related factor 2 (Nrf2) were substantially elevated with BCSE treatment, providing a direct evidence of a coordinated activation of the Nrf2-regulated antioxidant pathway, which led to the upregulation of a variety of housekeeping genes. The results of our study provide substantial evidence that black currant bioactive anthocyanins exert chemopreventive actions against DENA-inflicted hepatocarcinogenesis by attenuating oxidative stress through activation of Nrf2 signaling pathway.
Thoppil Roslin J; Bhatia Deepak; Barnes Kendra F; Haznagy-Radnai Erzsebet; Hohmann Judit; Darvesh Altaf S; Bishayee Anupam
Current cancer drug targets
2012
2012-11
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.2174/156800912803987968" target="_blank" rel="noreferrer noopener">10.2174/156800912803987968</a>
A class of genes in the HER2 regulon that is poised for transcription in breast cancer cell lines and expressed in human breast tumors.
Humans; Cell Line; *Gene Expression Regulation; Reverse Transcriptase Polymerase Chain Reaction; *Gene Expression Profiling; Breast Neoplasms/genetics/pathology; Gene Regulatory Networks; Homeodomain Proteins/genetics/metabolism; MCF-7 Cells; Nanog Homeobox Protein; Neoplastic Stem Cells/metabolism; Octamer Transcription Factor-3/genetics/metabolism; Regulon/*genetics; RNA Polymerase II/metabolism; SOXB1 Transcription Factors/genetics/metabolism; Tumor Microenvironment/genetics; Receptor; Blotting; Western; Tumor; Neoplastic; ErbB-2/*genetics/metabolism
HER2-positive breast cancer accounts for 25% of all cases and has a poor prognosis. Although progress has been made in understanding signal transduction, little is known of how HER2 achieves gene regulation. We performed whole genome expression analysis on a HER2(+) and HER2(-) breast cancer cell lines and compared these results to expression in 812 primary tumors stratified by their HER2 expression level. Chip-on-chip with anti-RNA polymerase II was compared among breast cancer cell lines to identify genes that are potentially activated by HER2. The expression levels of these HER2-dependent POL II binding genes were determined for the 812 HER2+/- breast cancer tissues. Genes differentially expressed between HER2+/- cell lines were generally regulated in the same direction as in breast cancer tissues. We identified genes that had POLII binding in HER2(+) cell lines, but without significant gene expression. Of 737 such genes "poised" for expression in cell lines, 113 genes were significantly differentially expressed in breast tumors in a HER2-dependent manner. Pathway analysis of these 113 genes revealed that a large group of genes were associated with stem cell and progenitor cell control as indicated by networks centered on NANOG, SOX2, OCT3/4. HER2 directs POL II binding to a large number of genes in breast cancer cells. A "poised" class of genes in HER2(+) cell lines with POLII binding and low RNA expression but is differentially expressed in primary tumors, strongly suggests a role of the microenvironment and further suggests a role for stem cells proliferation in HER2-regulated breast cancer tissue.
Rahmatpanah Farah B; Jia Zhenyu; Chen Xin; Char Jessica E; Men Bozhao; Franke Anna-Clara; Jones Frank E; McClelland Michael; Mercola Dan
Oncotarget
2015
2015-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.18632/oncotarget.2676" target="_blank" rel="noreferrer noopener">10.18632/oncotarget.2676</a>
Ambient GABA-activated tonic inhibition sharpens auditory coincidence detection via a depolarizing shunting mechanism.
Animals; Chick Embryo; Patch-Clamp Techniques; Electric Stimulation; Neurons/*physiology; gamma-Aminobutyric Acid/*physiology; Membrane Potentials/physiology; Auditory Pathways/*physiology; Neural Inhibition/*physiology; Inhibitory Postsynaptic Potentials; Receptors; Blotting; Western; GABA-A/*physiology
Tonic inhibition mediated by extrasynaptic GABA(A) receptors (GABA(A)Rs) has emerged as a novel form of neural inhibition in the CNS. However, little is known about its presence and function in the auditory system. Using whole-cell recordings in brain slices, we identified a tonic current mediated by GABA(A)Rs containing the delta subunit in middle/high-characteristic-frequency neurons of the chicken nucleus laminaris, the first interaural time difference encoder that computes information for sound localization. This tonic conductance was activated by ambient concentrations of GABA released from synaptic vesicles. Furthermore, pharmacological manipulations of the conductance demonstrated its essential role in coincidence detection. Remarkably, this depolarizing tonic conductance was strongly inhibitory primarily because of its shunting effect. These results demonstrate a novel role for tonic inhibition in central auditory information processing.
Tang Zheng-Quan; Dinh Emilie Hoang; Shi Wei; Lu Yong
The Journal of neuroscience : the official journal of the Society for Neuroscience
2011
2011-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1523/JNEUROSCI.4733-10.2011" target="_blank" rel="noreferrer noopener">10.1523/JNEUROSCI.4733-10.2011</a>
Impact of type 1 diabetes on cardiac fibroblast activation: enhanced cell cycle progression and reduced myofibroblast content in diabetic myocardium.
Animals; Blood Glucose/metabolism; Blotting; Body Weight/physiology; Cell Cycle Proteins/biosynthesis; Cell Cycle/*physiology; Cell Differentiation/physiology; Cell Proliferation; Cell Separation; Diabetes Mellitus; Echocardiography; Experimental/pathology; Fibroblasts/*physiology; Male; Microarray Analysis; Myocardium/cytology/*pathology; Myofibroblasts/*physiology; Phenotype; Rats; RNA/biosynthesis/isolation & purification; Signal Transduction/physiology; Sprague-Dawley; Type 1/diagnostic imaging/*pathology; Western
Diabetic patients are prone to developing myocardial fibrosis and suffer from decreased wound healing capabilities. The purpose of this study was to determine whether diabetes alters cardiac fibroblast activity in the myocardium in a 6-wk streptozotocin-induced type 1 diabetic model. In vivo echocardiography indicated significant dilation of the left ventricle (LV) in the diabetic animals, while cardiac function was comparable to that in the normal group. We isolated cardiac fibroblasts from diabetic and control hearts and observed increased proliferation of the diabetic fibroblasts. Microarray analysis using mRNA collected from whole LVs revealed downregulation of known inhibitors of proliferation, p53 and p21, in the diabetic group, consistent with our proliferation data. Western blot analysis confirmed a reduction in p53 protein expression in the diabetic hearts compared with control. We explored the potential signaling underlying the downregulation of these cell cycle mediators and determined that activated Akt, a signal that inhibits p53, was elevated in the diabetic group. Surprisingly, the hearts from the diabetic group contained lower levels of the myofibroblast marker alpha-smooth muscle actin (alpha-SMA) and higher levels of desmin and platelet endothelial cell adhesion molecule (PECAM). The isolated fibroblasts from the diabetic group also contained significantly less alpha-SMA. These data suggest that early-stage diabetic hearts contain highly proliferative fibroblasts, which predisposes the diabetic myocardium to fibrosis, but have fewer myofibroblasts, which may compromise wound healing.
Shamhart Patricia E; Luther Daniel J; Hodson Ben R; Koshy John C; Ohanyan Vahagn; Meszaros J Gary
American journal of physiology. Endocrinology and metabolism
2009
2009-11
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1152/ajpendo.00327.2009" target="_blank" rel="noreferrer noopener">10.1152/ajpendo.00327.2009</a>
Activation of metabotropic glutamate receptor 1 dimers requires glutamate binding in both subunits.
Blotting; Calcium Channels/drug effects/metabolism; DNA/biosynthesis/genetics; Dose-Response Relationship; Drug; Fluorescent Antibody Technique; Genes; Glutamic Acid/*metabolism; Humans; Membrane Potentials/drug effects; Metabotropic Glutamate/genetics/*metabolism; myc/genetics; Patch-Clamp Techniques; Plasmids/genetics; Receptors; Signal Transduction/drug effects; Superior Cervical Ganglion/cytology/drug effects/metabolism; Sympathetic Nervous System/cytology/drug effects/metabolism; Western
Group I metabotropic glutamate receptors (mGluRs) form stable, disulfide-linked homodimers. Lack of a verifiably monomeric mGluR1 mutant has led to difficulty in assessing the role of dimerization in the molecular mechanism of mGluR1 activation. The related GABA(B) receptor exhibits striking intradimer cross talk (ligand binding at one subunit effectively produces G protein activation at the other), but it is unclear whether group I mGluRs exhibit analogous cross talk. Signaling of heterologously expressed mGluR1 was examined in isolated rat sympathetic neurons by measuring glutamate-mediated inhibition of native calcium currents. To examine mGluR1 activity when only one dimer subunit has access to glutamate ligand, wildtype mGluR1 was coexpressed with mGluR1 Y74A, a mutant with impaired glutamate binding, and the activity of the heterodimer (mutant/wild type) was examined. The mGluR1 Y74A mutant alone had a dose-response curve that was shifted by about 2 orders of magnitude. The half-maximal dose of glutamate shifted from 1.3 (wild-type mGluR1) to about 450 (mGluR1 Y74A) microM. However, the maximal effect was similar. Wild-type mGluR1 was expressed with excess Y74A mGluR1 to generate a receptor population consisting largely of mutant homodimers and mutant/wild-type heterodimers but without detectable wild-type homodimers. Under these conditions, no glutamate-mediated calcium current inhibition was observed below approximately 300 microM glutamate, although wild-type mGluR1 protein was detectable with immunofluorescence. These data suggest that mutant/wild-type heterodimeric receptors are inactive at ligand concentrations favoring glutamate association with receptor dimers at only one subunit.
Kammermeier Paul J; Yun June
The Journal of pharmacology and experimental therapeutics
2005
2005-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1124/jpet.104.073155" target="_blank" rel="noreferrer noopener">10.1124/jpet.104.073155</a>
Rifampicin induction of CYP3A4 requires pregnane X receptor cross talk with hepatocyte nuclear factor 4alpha and coactivators, and suppression of small heterodimer partner gene expression.
Antibiotics; Antitubercular/*pharmacology; Blotting; Cell Line; Chromatin/metabolism; Cytochrome P-450 CYP3A; Cytochrome P-450 Enzyme System/*biosynthesis; Cytoplasmic and Nuclear/*biosynthesis/*drug effects/genetics; Electrophoretic Mobility Shift Assay; Enzyme Induction/drug effects; Glutathione Transferase/metabolism; Hepatocyte Nuclear Factor 4/genetics/*metabolism; Hepatocytes/drug effects/metabolism; Humans; Immunoprecipitation; Messenger/biosynthesis; Plasmids/genetics; Pregnane X Receptor; Receptor Cross-Talk/drug effects; Receptors; Reverse Transcriptase Polymerase Chain Reaction; Rifampin/*pharmacology; RNA; Steroid/*drug effects/genetics; Transfection; Tumor; Western
Bile acids and drugs activate pregnane X receptor (PXR) to induce CYP3A4, which is the predominant cytochrome P450 enzyme expressed in the liver and intestine and plays a critical role in detoxifying bile acids and drugs, and protecting against cholestasis. The aim of this study is to investigate the molecular mechanism of PXR cross talk with other nuclear receptors and coactivators in regulating human CYP3A4 gene transcription. Rifampicin dose dependently induced the CYP3A4 but inhibited small heterodimer partner (SHP) mRNA expression levels in primary human hepatocytes. Rifampicin strongly stimulated PXR and hepatocyte nuclear factor 4alpha (HNF4alpha) interaction, and CYP3A4 reporter activity, which was further stimulated by peroxisome proliferators-activated receptorgamma co-activator 1alpha (PGC-1alpha) and steroid receptor coactivator-1 (SRC-1) but inhibited by SHP. Mutation of the putative HNF4alpha binding site in the distal xenobiotic responsive element module did not affect CYP3A4 basal promoter activity and synergistic stimulation by PXR and HNF4alpha. Chromatin immunoprecipitation assays revealed that rifampicin-activated PXR recruited HNF4alpha and SRC-1 to the CYP3A4 chromatin. On the other hand, SHP reduced PXR recruitment of HNF4alpha and SRC-1 to the CYP3A4 chromatin. The human SHP promoter was stimulated by HNF4alpha and PGC-1alpha. Upon activation by rifampicin, PXR inhibited SHP promoter activity. Results suggest that PXR strongly induces CYP3A4 gene transcription by interacting with HNF4alpha, SRC-1, and PGC-1alpha. PXR concomitantly inhibits SHP gene transcription and maximizes the PXR induction of the CYP3A4 gene in human livers. Drugs targeted to PXR may be developed for treating cholestatic liver diseases induced by bile acids and drugs.
Li Tiangang; Chiang John Y L
Drug metabolism and disposition: the biological fate of chemicals
2006
2006-05
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1124/dmd.105.007575" target="_blank" rel="noreferrer noopener">10.1124/dmd.105.007575</a>
Pomegranate-mediated chemoprevention of experimental hepatocarcinogenesis involves Nrf2-regulated antioxidant mechanisms.
*Phytotherapy; Alkylating Agents/toxicity; Animals; Antioxidants/*therapeutic use; Blotting; Carcinoma; Diethylnitrosamine/toxicity; Experimental/chemically induced/*drug therapy/metabolism; GA-Binding Protein Transcription Factor/*metabolism; gamma-Glutamyltransferase/metabolism; Hepatocellular/chemically induced/*drug therapy/metabolism; Immunoenzyme Techniques; Lipid Peroxidation/drug effects; Liver Neoplasms; Male; Oxidative Stress; Plant Extracts/*therapeutic use; Punicaceae/*chemistry; Rats; Sprague-Dawley; Western
Hepatocellular carcinoma (HCC), one of the most prevalent and lethal cancers, has shown an alarming rise in the USA. Without effective therapy for HCC, novel chemopreventive strategies may effectively circumvent the current morbidity and mortality. Oxidative stress predisposes to hepatocarcinogenesis and is the major driving force of HCC. Pomegranate, an ancient fruit, is gaining tremendous attention due to its powerful antioxidant properties. Here, we examined mechanism-based chemopreventive potential of a pomegranate emulsion (PE) against dietary carcinogen diethylnitrosamine (DENA)-induced rat hepatocarcinogenesis that mimics human HCC. PE treatment (1 or 10 g/kg), started 4 weeks prior to the DENA challenge and continued for 18 weeks thereafter, showed striking chemopreventive activity demonstrated by reduced incidence, number, multiplicity, size and volume of hepatic nodules, precursors of HCC. Both doses of PE significantly attenuated the number and area of gamma-glutamyl transpeptidase-positive hepatic foci compared with the DENA control. PE also attenuated DENA-induced hepatic lipid peroxidation and protein oxidation. Mechanistic studies revealed that PE elevated gene expression of an array of hepatic antioxidant and carcinogen detoxifying enzymes in DENA-exposed animals. PE elevated protein and messenger RNA expression of the hepatic nuclear factor
Bishayee Anupam; Bhatia Deepak; Thoppil Roslin J; Darvesh Altaf S; Nevo Eviatar; Lansky Ephraim P
Carcinogenesis
2011
2011-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1093/carcin/bgr045" target="_blank" rel="noreferrer noopener">10.1093/carcin/bgr045</a>
Altered Neuroinflammation and Behavior after Traumatic Brain Injury in a Mouse Model of Alzheimer's Disease.
Alzheimer Disease/etiology/*metabolism/pathology; Alzheimer's disease; Amyloid beta-Peptides/*metabolism; Animal; Animal/physiology; Animals; Behavior; Blotting; Brain Injuries; Brain/*metabolism/pathology; Disease Models; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Humans; Immunohistochemistry; Inbred C57BL; Inflammation/*metabolism/pathology; macrophage; Mice; neuroinflammation; Transgenic; traumatic brain injury; Traumatic/complications/*metabolism/*pathology; Western
Traumatic brain injury (TBI) has acute and chronic sequelae, including an increased risk for the development of Alzheimer's disease (AD). TBI-associated neuroinflammation is characterized by activation of brain-resident microglia and infiltration of monocytes; however, recent studies have implicated beta-amyloid as a major manipulator of the inflammatory response. To examine neuroinflammation after TBI and development of AD-like features, these studies examined the effects of TBI in the presence and absence of beta-amyloid. The R1.40 mouse model of cerebral amyloidosis was used, with a focus on time points well before robust AD pathologies. Unexpectedly, in R1.40 mice, the acute neuroinflammatory response to TBI was strikingly muted, with reduced numbers of CNS myeloid cells acquiring a macrophage phenotype and decreased expression of inflammatory cytokines. At chronic time points, macrophage activation substantially declined in non-Tg TBI mice; however, it was relatively unchanged in R1.40 TBI mice. The persistent inflammatory response coincided with significant tissue loss between 3 and 120 days post-injury in R1.40 TBI mice, which was not observed in non-Tg TBI mice. Surprisingly, inflammatory cytokine expression was enhanced in R1.40 mice compared with non-Tg mice, regardless of injury group. Although R1.40 TBI mice demonstrated task-specific deficits in cognition, overall functional recovery was similar to non-Tg TBI mice. These findings suggest that accumulating beta-amyloid leads to an altered post-injury macrophage response at acute and chronic time points. Together, these studies emphasize the role of post-injury neuroinflammation in regulating long-term sequelae after TBI and also support recent studies implicating beta-amyloid as an immunomodulator.
Kokiko-Cochran Olga N; Ransohoff Lena; Veenstra Mike; Lee Sungho; Saber Maha; Sikora Matt; Teknipp Ryan; Xu Guixiang; Bemiller Shane M; Wilson Gina; Crish Samuel; Bhaskar Kiran; Lee Yu-Shang; Ransohoff Richard M; Lamb Bruce T
Journal of neurotrauma
2016
2016-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1089/neu.2015.3970" target="_blank" rel="noreferrer noopener">10.1089/neu.2015.3970</a>
Bcl-2 positively regulates Sox9-dependent chondrocyte gene expression by suppressing the MEK-ERK1/2 signaling pathway.
*Gene Expression Regulation; Adenoviridae/genetics; Animals; Apoptosis; beta-Galactosidase/metabolism; Blotting; Butadienes/pharmacology; Caspase Inhibitors; Cell Differentiation; Cell Line; Chondrocytes/*metabolism; Collagen Type II/metabolism; Down-Regulation; Enzyme Inhibitors/pharmacology; Fibroblasts/metabolism; Fluorescence; Genetic; High Mobility Group Proteins/*metabolism; Lac Operon; Luciferases/metabolism; MAP Kinase Kinase Kinases/*metabolism; Messenger/metabolism; Microscopy; Mitogen-Activated Protein Kinase 1/*metabolism; Mitogen-Activated Protein Kinase 3/*metabolism; NF-kappa B/metabolism; Nitriles/pharmacology; Phenotype; Phosphorylation; Promoter Regions; Protein Kinase C-alpha; Protein Kinase C/antagonists & inhibitors; Proteoglycans/metabolism; Proto-Oncogene Proteins c-bcl-2/*metabolism; Rats; Reverse Transcriptase Polymerase Chain Reaction; RNA; Signal Transduction; Small Interfering/metabolism; SOX9 Transcription Factor; Sprague-Dawley; Time Factors; Transcription; Transcription Factors/*metabolism; Transfection; Western
Bcl-2 is an anti-apoptotic protein that has recently been shown to regulate other cellular functions. We previously reported that Bcl-2 regulates chondrocyte matrix gene expression, independent of its anti-apoptotic function. Here, we further investigate this novel function of Bcl-2 and examine three intracellular signaling pathways likely to be associated with this function. The present study demonstrates that the activity of Sox9, a master transcription factor that regulates the gene expression of chondrocyte matrix proteins, is suppressed by Bcl-2 small interference RNA in the presence of caspase inhibitors. This effect was attenuated by prior exposure of chondrocytes to an adenoviral vector expressing sense Bcl-2. In addition, the down-regulation of Bcl-2, Sox9, and chondrocyte-specific gene expression by serum withdrawal in primary chondrocytes was reversed by expressing Bcl-2. Inhibition of the protein kinase C alpha and NFkappaB pathways had no effect on the maintenance of Sox9-dependent gene expression by Bcl-2. In contrast, whereas the MEK-ERK1/2 pathway negatively regulated the differentiated phenotype in wild type chondrocytes, inhibition of this pathway reversed the loss of differentiation markers and fibroblastic phenotype in Bcl-2-deficient chondrocytes. In conclusion, the present study identifies a specific signaling pathway, namely, MEK-ERK1/2, that is downstream of Bcl-2 in the regulation of Sox9-dependent chondrocyte gene expression and phenotype.
Yagi Rieko; McBurney Denise; Horton Walter E Jr
The Journal of biological chemistry
2005
2005-08
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1074/jbc.M502751200" target="_blank" rel="noreferrer noopener">10.1074/jbc.M502751200</a>
Phosphatidylinositide 3-kinase regulates angiotensin II-induced cytosolic phospholipase A2 activity and growth in vascular smooth muscle cells.
Angiotensin II/*metabolism; Animals; Arachidonic Acid/metabolism; Arachidonic Acids/pharmacology; Blotting; Cells; Chromones/pharmacology; Cultured; Enzyme Inhibitors/pharmacology; Flavonoids/pharmacology; Group IV Phospholipases A2; Male; Mitogen-Activated Protein Kinases/metabolism; Morpholines/pharmacology; Muscle; Phosphatidylinositol 3-Kinases/*physiology; Phospholipases A/*metabolism; Phospholipases A2; Phosphorylation; Rats; Smooth; Sprague-Dawley; Vascular/drug effects/*growth & development; Western
Angiotensin (Ang) II via the AT(1) receptor acts as a mitogen in vascular smooth muscle cells (VSMC) through stimulation of multiple signaling mechanisms, including tyrosine kinases and mitogen-activated protein kinase (MAPK). In addition, cytosolic phospholipase A(2)(cPLA(2))-dependent release of arachidonic acid (AA) is linked to VSMC growth and we have reported that Ang II stimulates cPLA(2) activity via the AT(1) receptor. The coupling of Ang II to the activation of cPLA(2) appears to involve mechanisms both upstream and downstream of MAPK such that AA stimulates MAPK activity which phosphorylates cPLA(2) to further enhance AA release. However, the upstream mechanisms responsible for activation of cPLA(2) are not well-defined. One possibility includes phosphatidylinositide
Silfani Tonous N; Freeman Ernest J
Archives of biochemistry and biophysics
2002
2002-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/S0003-9861(02)00066-8" target="_blank" rel="noreferrer noopener">10.1016/S0003-9861(02)00066-8</a>
Markers associated with testosterone enhancement of methamphetamine-induced striatal dopaminergic neurotoxicity.
Animals; Biomarkers/metabolism; Blotting; Body Temperature/drug effects; Body Weight/drug effects; Corpus Striatum/*drug effects/metabolism; Dopamine Plasma Membrane Transport Proteins/metabolism; Dopamine/*metabolism; Drug Synergism; HSP70 Heat-Shock Proteins/metabolism; Inbred Strains; Male; Methamphetamine/*toxicity; Mice; Neurotoxicity Syndromes/*etiology/metabolism; Oxidative Stress/drug effects; Testosterone Propionate/*pharmacology; Vesicular Monoamine Transport Proteins/metabolism; Western
Intact male CD-1 mice received an injection of testosterone propionate (TP–5 ug), progesterone (P–5 mg), the oil vehicle or remained untreated (control). At 24 hours after hormonal treatments the mice received an injection of methamphetamine (MA–40 mg/kg) and rectal temperatures were measured. At 5 days post-MA, assays were performed to assess effects of these treatments. Maximal increases in body temperatures, that were significantly greater than oil-treated controls, were obtained in TP-treated mice. At 5 days post-MA, maximal weight reductions were obtained with TP-treated mice, while P-treated mice showed no significant decrease between the pre- versus post-MA determinations. Striatal dopamine concentrations showed maximal reductions and heat-shock protein-70 maximal increases in the TP group, with both differing significantly as compared with all other groups. Protein levels of dopamine transporters were significantly decreased in P-treated mice, while vesicular monoamine transporter-2 was significantly decreased in TP-treated mice. Taken together, these results suggest that testosterone exacerbates the deleterious effects of MA within male mice as indicated by a number of markers related to neurotoxicity. The changes in markers as associated with this enhanced neurotoxicity suggest that TP may increase thermal/energy responses and/or oxidative stress to produce this effect.
Buletko A Blake; Dluzen Dean E; McDermott Janet L; Darvesh Altaf S; Geldenhuys Werner J
Neurotoxicology and teratology
2012
2012-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.ntt.2012.03.003" target="_blank" rel="noreferrer noopener">10.1016/j.ntt.2012.03.003</a>
Resveratrol inhibition of varicella-zoster virus replication in vitro.
Antiviral Agents/*pharmacology; Blotting; Cell Line; Dose-Response Relationship; Drug; Fibroblasts/virology; Herpesvirus 3; Human/*drug effects/physiology; Humans; Immediate-Early Proteins/biosynthesis; Messenger/biosynthesis; Resveratrol; Reverse Transcriptase Polymerase Chain Reaction; RNA; Stilbenes/*pharmacology; Trans-Activators/biosynthesis; Viral Envelope Proteins/biosynthesis; Viral Plaque Assay; Viral/biosynthesis; Virus Attachment/drug effects; Virus Inactivation/drug effects; Virus Replication/*drug effects; Western
Resveratrol was found to inhibit varicella-zoster virus (VZV) replication in a dose-dependent and reversible manner. This decrease in virus production in the presence of resveratrol was not caused by direct inactivation of VZV or inhibition of virus attachment to MRC-5 cells. The drug effectively limited VZV replication if added during the first 30 h of infection. Western blot analysis and real-time RT-PCR studies demonstrated that protein and mRNA levels of IE62, an essential immediate early viral protein, were reduced when compared to controls. These results demonstrate that VZV replication is adversely affected by resveratrol which is negatively impacting IE62 synthesis.
Docherty John J; Sweet Thomas J; Bailey Erin; Faith Seth A; Booth Tristan
Antiviral research
2006
2006-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.antiviral.2006.07.004" target="_blank" rel="noreferrer noopener">10.1016/j.antiviral.2006.07.004</a>
Histone Deacetylase Inhibitor Vorinostat (SAHA) Suppresses IL-1beta-Induced Matrix Metallopeptidase-13 Expression by Inhibiting IL-6 in Osteoarthritis Chondrocyte.
Aged; Articular/drug effects/metabolism; Blotting; Cartilage; Chondrocytes/drug effects; Collagen Type II/drug effects/metabolism; Down-Regulation/drug effects; Female; Gene Expression Regulation/*drug effects; Histone Deacetylase Inhibitors/*pharmacology; Humans; Hydroxamic Acids/*pharmacology; Interleukin-1beta/*antagonists & inhibitors/genetics/metabolism; Knee Joint/metabolism; Male; Matrix Metalloproteinase 13/*drug effects/metabolism; Middle Aged; Osteoarthritis/*drug therapy; Proteoglycans/metabolism; Tumor Necrosis Factor-alpha/drug effects/metabolism; Vorinostat; Western
Osteoarthritis (OA) is the most common whole-joint disease and is characterized by progressive loss of the cartilage matrix. Matrix metallopeptidase-13 (MMP-13) is a highly active and an abundantly expressed protease in OA cartilage and chondrocytes and degrades type II collagen and proteoglycans. We investigated the mechanism of MMP-13 suppression by histone deacetylase inhibitor vorinostat (SAHA). OA chondrocytes were obtained from knee cartilage after enzymatic digestion and treated with IL-1beta in the absence or presence of various histone deacetylase inhibitors. Gene expression was quantified using quantitative RT-PCR. Protein expression and chromatin modifications were determined by Western immunoblotting using specific antibodies. The effect of IL-6 on the expression of
Makki Mohammad Shahidul; Haqqi Tariq M
The American journal of pathology
2016
2016-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.ajpath.2016.06.010" target="_blank" rel="noreferrer noopener">10.1016/j.ajpath.2016.06.010</a>
The Detrimental Role Played by Lipocalin-2 in Alcoholic Fatty Liver in Mice.
Alcoholic/*metabolism; Animal; Animals; Blotting; Disease Models; Fatty Liver; Humans; Inbred C57BL; Knockout; Lipocalin-2/*metabolism; Mice; Polymerase Chain Reaction; Western
We have previously shown that the ethanol-mediated elevation of lipocaline-2 (LCN2) is closely associated with the development of alcoholic fatty liver disease (AFLD) in mice. Herein, we aimed to understand the functional significance of LCN2 induction by ethanol and to explore its underlying mechanisms. We evaluated the effects of LCN2 in an in vitro cellular alcoholic steatosis model and in an animal study using wild-type and LCN2 knockout mice fed for 4 weeks with an ethanol-supplemented Lieber-DeCarli diet. In the cellular model of alcoholic steatosis, recombinant LCN2 or overexpression of LCN2 exacerbated ethanol-induced fat accumulation, whereas knocking down LCN2 prevented steatosis in hepatocytes exposed to ethanol. Consistently, removal of LCN2 partially but significantly alleviated alcoholic fatty liver injury in mice. Mechanistically, LCN2 mediates detrimental effects of ethanol in the liver via disrupted multiple signaling pathways, including aberrant nicotinamide phosphoribosyltransferase-sirtuin 1 axis, perturbed endocrine metabolic regulatory fibroblast growth factor 15/19 signaling, and impaired chaperone-mediated autophagy. Finally, compared with healthy human livers, liver samples from patients with AFLD had lower gene expression of several
Cai Yan; Jogasuria Alvin; Yin Huquan; Xu Ming-Jiang; Hu Xudong; Wang Jiayou; Kim Chunki; Wu Jiashin; Lee Kwangwon; Gao Bin; You Min
The American journal of pathology
2016
2016-09
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.ajpath.2016.05.006" target="_blank" rel="noreferrer noopener">10.1016/j.ajpath.2016.05.006</a>
Salvianolate reduces murine myocardial ischemia and reperfusion injury via ERK1/2 signaling pathways in vivo.
*MAP Kinase Signaling System/drug effects; Animal Studies; Animals; Blotting; Cardiotonic Agents/pharmacology/therapeutic use; Chinese medicine; Descriptive Statistics; extracellular signal-regulated kinase 1/2; Flavonoids/pharmacology; Heart Ventricle; Heart Ventricles/drug effects/pathology; In Vivo Studies; Inbred C57BL; ischemia and reperfusion injury; Left; Male; Mice; Mitogen-Activated Protein Kinase 1/*metabolism; Mitogen-Activated Protein Kinase 3/*metabolism; Molecular Structure; Myocardial Ischemia – Drug Therapy; Myocardial Reperfusion Injury – Drug Therapy; Myocardial Reperfusion Injury/*drug therapy/enzymology/pathology; Organ Size/drug effects; P-Value; Phosphorylation; Phosphorylation/drug effects; Plant Extracts – Administration and Dosage; Plant Extracts/chemistry/pharmacology/*therapeutic use; protein kinase B; Protein Kinase Inhibitors – Administration and Dosage; Protein Kinase Inhibitors/pharmacology; Protein Kinases – Analysis; Protein Kinases – Drug Effects; salvianolate; Signal Transduction; Staining and Labeling; Western
OBJECTIVE: To analyze the effects of salvianolate on myocardial infarction in a murine in vivo model of ischemia and reperfusion (I/R) injury. METHODS: Myocardial I/R injury model was constructed in mice by 30 min of coronary occlusion followed by 24 h of reperfusion and pretreated with salvianolate 30 min before I/R (SAL group). The SAL group was compared with SHAM (no I/R and no salvianolate), I/R (no salvianolate), and ischemia preconditioning (IPC) groups. Furthermore, an ERK1/2 inhibitor PD98059 (1 mg/kg), and a phosphatidylinositol-3-kinase (PI3-K) inhibitor, LY294002 (7.5 mg/kg), were administered intraperitoneal injection (i.p) for 30 min prior to salvianolate, followed by I/R surgery in LY and PD groups. By using a double staining method, the ratio of the infarct size (IS) to left ventricle (LV) and of risk region (RR) to LV were compared among the groups. Correlations between IS and RR were analyzed. Western-blot was used to detect the extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (AKT) phosphorylation changes. RESULTS: There were no significant differences between RR to LV ratio among the SHAM, I/R, IPC and SAL groups (P\textgreater0.05). The SAL and IPC groups had IS of 26.1%+/-1.4% and 22.3%+/-2.9% of RR, respectively, both of which were significantly smaller than the I/R group (38.5%+/-2.9% of RR, P\textless0.05, P\textless0.01, respectively). Moreover, the phosphorylation of ERK1/2 was increased in SAL group (P\textless0.05), while AKT had no significant change. LY294002 further reduced IS, whereas the protective role of salvianolate could be attenuated by PD98059, which increased the IS. Additionally, the IS was not linearly related to the RR (r=0.23, 0.45, 0.62, 0.17, and 0.52 in the SHAM, I/R, SAL, LY and PD groups, respectively). CONCLUSION: Salvianolate could reduce myocardial I/R injury in mice in vivo, which involves an ERK1/2 pathway, but not a PI3-K signaling pathway.
Qi Jianyong; Yu Juan; Huang Dong-Hui; Guo Liheng; Wang Lei; Huang Xin; Huang Hai-Ding; Zhou Miao; Zhang Minzhou; Wu Jiashin
Chinese journal of integrative medicine
2017
2017-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s11655-016-2621-z" target="_blank" rel="noreferrer noopener">10.1007/s11655-016-2621-z</a>
Chemopreventive doses of resveratrol do not produce cardiotoxicity in a rodent model of hepatocellular carcinoma.
*Chemoprevention; Analysis of Variance; Animal; Animal Studies; Animal/drug effects; Animals; Antioxidants; Behavior; Blotting; Carcinoma; Cardiotoxicity; Cardiotoxins/*toxicity; Chemoprevention; Data Analysis Software; Descriptive Statistics; Disease Models; Doppler; Dose-Response Relationship; Drug; Echocardiography; Feeding Behavior/drug effects; Female; Fisher's Exact Test; Funding Source; Heart – Drug Effects; Heart/drug effects/physiopathology; Hepatocellular – Prevention and Control; Hepatocellular/*drug therapy/pathology/physiopathology; Hepatocytes/drug effects/pathology; Humans; Liver Neoplasms/*drug therapy/pathology/physiopathology; Liver/drug effects/pathology/physiopathology; Polyphenols – Therapeutic Use; Rats; Resveratrol; Sprague-Dawley; Stilbenes/*therapeutic use; Systole/drug effects; Western
Hepatocellular carcinoma (HCC), one of the most lethal cancers, results in more than one million fatalities worldwide every year. In view of the limited therapeutic alternatives and poor prognosis of liver cancer, preventive control approaches, notably chemoprevention, have been considered to be the best strategy in lowering the present prevalence of the disease. Resveratrol, a naturally occurring antioxidant and antiinflammatory agent found in grapes and red wine, inhibits carcinogenesis with a pleiotropic mode of action. Recently, we have reported that dietary resveratrol significantly prevents chemically-induced liver tumorigenesis in rats. One of the mechanisms of resveratrol-mediated chemoprevention of hepatocarcinogenesis could be related to its antiinflammatory action through hepatic cyclooxygenase (COX-2) inhibition. Although several COX-2 inhibitors are known to exert chemopreventive efficacy, not all are considered ideal candidates for chemoprevention due to the risk of adverse cardiovascular events. Accordingly, the objective of the present study was to evaluate the role of resveratrol on cardiac performance during experimental hepatocarcinogenesis initiated with diethylnitrosamine and promoted by phenobarbital. Rats had free access to diet supplemented with resveratrol four weeks before the carcinogen injection and 14 weeks thereafter. The cardiotoxicity of resveratrol was assessed by monitoring the cardiac function using transthoracic echocardiography as well as Western blot analysis of cardiac tissue. Long-term dietary administration of resveratrol dose-dependently suppressed hepatic tumor multiplicity, the principal endpoint for evaluating the chemopreventive potential of a candidate agent. The chemopreventive effects of resveratrol were also reflected in histopathological assessment of hepatic tissues. Resveratrol did not exhibit any cardiotoxicity but rather improved the cardiac function in a dose-responsive fashion. Our results indicate that resveratrol-mediated chemoprevention of rat liver carcinogenesis is devoid of any adverse cardiovascular events. Resveratrol may be developed as a chemopreventive as well as therapeutic drug for human HCC.
Luther Daniel J; Ohanyan Vahagn; Shamhart Patricia E; Hodnichak Cheryl M; Sisakian Hamayak; Booth Tristan D; Meszaros J Gary; Bishayee Anupam
Investigational new drugs
2011
2011-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s10637-009-9332-7" target="_blank" rel="noreferrer noopener">10.1007/s10637-009-9332-7</a>
P2X receptors in the rat uterine cervix, lumbosacral dorsal root ganglia, and spinal cord during pregnancy.
Animals; Blotting; Cervix Uteri/*metabolism; Estrogen Receptor alpha/metabolism; Female; Frozen Sections; Ganglia; Immunohistochemistry; Lumbosacral Region; Pregnancy; Purinergic P2/*metabolism; Purinergic P2X; Rats; Receptors; Spinal Cord/cytology/*metabolism; Spinal/cytology/*metabolism; Sprague-Dawley; Western
ATP, an intracellular energy source, is released from cells during tissue stress, damage, or inflammation. The P2X subtype of the ATP receptor is expressed in rat dorsal root ganglion (DRG) cells, spinal cord dorsal horn, and axons in peripheral tissues. ATP binding to P2X receptors on nociceptors generates signals that can be interpreted as pain from damaged tissue. We have hypothesized that tissue stress or damage in the uterine cervix during late pregnancy and parturition can lead to ATP release and sensory signaling via P2X receptors. Consequently, we have examined sensory pathways from the cervix in nonpregnant and pregnant rats for the presence of purinoceptors. Antiserum against the
Papka Raymond E; Hafemeister Jen; Storey-Workley Megan
Cell and tissue research
2005
2005-07
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s00441-005-1114-8" target="_blank" rel="noreferrer noopener">10.1007/s00441-005-1114-8</a>
Identification of a new P450 subfamily, CYP4F1, expressed in rat hepatic tumors.
2-Acetylaminofluorene; Aflatoxin B1; Amino Acid; Amino Acid Sequence; Animals; Antisense Elements (Genetics); Base Sequence; Blotting; Clofibrate; Cytochrome P-450 Enzyme System/analysis/*genetics; Humans; Liver Neoplasms/chemically induced/*enzymology/genetics; Male; Microsomes/enzymology; Molecular Sequence Data; Multigene Family; Northern; Rats; Sequence Homology; Western
The expression of the rat cytochrome P450 CYP4 family was studied in hepatic tumors. In most of the primary and transplantable hepatic tumors studied, lauric acid omega-hydroxylase activity associated with the CYP4A subfamily enzymes decreased. The expression of CYP4A proteins and mRNAs in these tumors as assessed by Western and Northern blot was undetectable. However, while RNA analysis revealed the absence of 4A1, 4A2, and 4A3 mRNAs, the expression of CYP4 gene(s) was detected. A Uni-ZAP cDNA library was constructed from a
Chen L; Hardwick J P
Archives of biochemistry and biophysics
1993
1993-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1006/abbi.1993.1003" target="_blank" rel="noreferrer noopener">10.1006/abbi.1993.1003</a>
Estrogen receptor-alpha and neural circuits to the spinal cord during pregnancy.
Afferent/*metabolism; Animal/*physiology; Animals; Autonomic/anatomy & histology/metabolism; Blotting; Estrogen Receptor alpha; Estrogen/*biosynthesis; Estrogens/*physiology; Female; Ganglia; Immunohistochemistry; Neurons; Parturition/physiology; Pregnancy; Proto-Oncogene Proteins c-fos/*biosynthesis; Rats; Receptors; Sensory/anatomy & histology/metabolism; Spinal Cord/anatomy & histology/metabolism; Sprague-Dawley; Time Factors; Uterus/innervation; Western
Estrogen receptor immunoreactivity and mRNAs are present in spinal cord neurons in locations that are associated with sensory and autonomic innervation of female reproductive organs. The present study was undertaken to examine the expression of estrogen receptor-alpha in the spinal cord during different stages of pregnancy and to determine whether estrogen receptor-alpha-expressing neurons are related to uterine afferent nerves bringing information to the spinal cord at parturition. Immunohistochemistry showed estrogen receptor-alpha-immunoreactive neurons in the dorsal one-half of the spinal cord, i.e., dorsal horn, dorsal intermediate gray areas (dorsal commissural nucleus), and around the central canal and sacral parasympathetic autonomic nucleus of the lumbosacral spinal cord. Neurons in these areas corresponded topographically to the distribution of central processes of visceral primary afferent neurons (e.g., containing calcitonin gene-related peptide and substance P) that innervate and activate second-order spinal cord neurons (evidenced by their expression of Fos) at parturition. Western blots showed that estrogen receptor-alpha increases in the spinal cord, with a peak at day 20 of gestation, followed by a slight decrease by 2 days postpartum. These studies show that estrogen receptor-alpha is expressed by neurons in autonomic and sensory areas of the lumbosacral spinal cord that have connections with the female reproductive system and that the level of estrogen receptor-alpha changes over the course of pregnancy, which may follow profiles of steroid hormones. Many of these neurons may be involved in processing information related to reproductive organ function, changes during pregnancy, and relays to other CNS centers.
Papka R E; Hafemeister J; Puder B A; Usip S; Storey-Workley M
Journal of neuroscience research
2002
2002-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/jnr.10421" target="_blank" rel="noreferrer noopener">10.1002/jnr.10421</a>
Comparative binding of biotinylated neurotrophins to alpha(2)-macroglobulin family of proteins: relationship between cytokine-binding and neuro-modulatory activities of the macroglobulins.
*Biotinylation; *Blotting; alpha-Macroglobulins/chemistry/*metabolism/pharmacology; Animals; Cytokines/chemistry/*metabolism; Humans; Iodine Radioisotopes/chemistry; Nerve Growth Factor/chemistry/metabolism; Nerve Growth Factors/chemistry/*metabolism; Neurons/drug effects; Pregnancy Proteins/chemistry/metabolism; Protein Binding/physiology; Rats; Species Specificity; Western
Human alpha(2)-macroglobulin (alpha(2)M), pregnancy zone protein (PZP), rat alpha(1)M and acute-phase rat alpha(2)M belong to the alpha(2)M gene family of proteins, which can react covalently with nucleophilic monoamines to yield monoamine-activated (MA) macroglobulins. The MA forms of human alpha(2)M, PZP and rat alpha(2)M have been demonstrated previously to inhibit various neurotrophin-promoted neuronal activities, whereas MA-alpha(1)M is neurostimulatory and all native macroglobulins are generally inactive. The mechanism of neuromodulation is unknown, but it has been postulated that MA macroglobulins might inhibit neurons via their binding and sequestration of neurotrophins. This study employed a novel biotinylation-Western blot technique to compare the neurotrophin-binding properties of the four macroglobulins, and to correlate their binding activities with their known neuro-modulatory activities. In comparison with their respective native counterparts, human and rat
Skornicka Erin L; Shi Xiaoqing; Koo Peter H
Journal of neuroscience research
2002
2002-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/jnr.10097" target="_blank" rel="noreferrer noopener">10.1002/jnr.10097</a>
Primary cilia regulates the directional migration and barrier integrity of endothelial cells through the modulation of hsp27 dependent actin cytoskeletal organization.
*Cell Movement; Actin Cytoskeleton/*metabolism; Animals; Blotting; Capillary Permeability/*physiology; Cell Adhesion; Cilia/metabolism; Endothelial Cells/cytology/*metabolism; Fluorescent Antibody Technique; Focal Adhesions/metabolism; HSP27 Heat-Shock Proteins/*metabolism; Mice; Polycystic Kidney Diseases/physiopathology; Signal Transduction/physiology; Transgenic; Western
Cilia are mechanosensing organelles that communicate extracellular signals into intracellular responses. Altered functions of primary cilia play a key role in the development of various diseases including polycystic kidney disease. Here, we show that endothelial cells from the oak ridge polycystic kidney (Tg737(orpk/orpk) ) mouse, with impaired cilia assembly, exhibit a reduction in the actin stress fibers and focal adhesions compared to wild-type (WT). In contrast, endothelial cells from polycystin-1 deficient mice (pkd1(null/null) ), with impaired cilia function, display robust stress fibers, and focal adhesion assembly. We found that the Tg737(orpk/orpk) cells exhibit impaired directional migration and endothelial cell monolayer permeability compared to the WT and pkd1(null/null) cells. Finally, we found that the expression of heat shock protein 27 (hsp27) and the phosphorylation of focal adhesion kinase (FAK) are downregulated in the Tg737(orpk/orpk) cells and overexpression of hsp27 restored both FAK phosphorylation and cell migration. Taken together, these results demonstrate that disruption of the primary cilia structure or function compromises the endothelium through the suppression of hsp27 dependent actin organization and focal adhesion formation, which may contribute to the vascular dysfunction in ciliopathies.
Jones Thomas J; Adapala Ravi K; Geldenhuys Werner J; Bursley Chris; AbouAlaiwi Wissam A; Nauli Surya M; Thodeti Charles K
Journal of cellular physiology
2012
2012-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/jcp.22704" target="_blank" rel="noreferrer noopener">10.1002/jcp.22704</a>
The presence of extracellular matrix alters the chondrocyte response to endoplasmic reticulum stress.
*Stress; Animals; Apoptosis/drug effects; Articular/cytology; Blotting; Cartilage; Cattle; Cells; Chondrocytes/cytology/drug effects/*metabolism; Cultured; DNA-Binding Proteins/genetics/metabolism; Dose-Response Relationship; Drug; Endoplasmic Reticulum/*metabolism; Extracellular Matrix/*metabolism; Glucose/pharmacology; Heat-Shock Proteins/genetics/metabolism; Physiological; Proto-Oncogene Proteins c-bcl-2/genetics/metabolism; Reverse Transcriptase Polymerase Chain Reaction; Thapsigargin/pharmacology; Time Factors; Transcription Factors/genetics/metabolism; Tunicamycin/pharmacology; Western
The objective of this study was to test the hypothesis that extracellular matrix (ECM) would alter the endoplasmic reticulum (ER) stress response of chondrocytes. Chondrocytes were isolated from calf knees and maintained in monolayer culture or suspended in collagen I to form spot cultures (SCs). Our laboratory has shown that bovine chondrocytes form cartilage with properties similar to native cartilage after 2-4 weeks in SCs. Monolayer cultures treated with ER stressors glucose withdrawal (-Glu), tunicamycin (TN), or thapsigargin (TG) up-regulated Grp78 and Gadd153, demonstrating a complete ER stress response. SCs were grown at specific times from 1 day to 6 weeks before treatment with ER stressors. Additionally, SCs grown for 1, 2, or 6 weeks were treated with increasing concentrations of TN or TG. Western blotting of SCs for Grp78 indicated that increased ECM accumulation results in delayed expression; however, Grp78 mRNA is up-regulated in response to ER stressors even after 6 weeks in culture. SCs treated with ER stressors did not up-regulate Gadd153, suggesting that the cells experienced ER stress but would not undergo apoptosis. In fact, SCs undergo apoptosis upon ER stress treatment after 0-1 day of growth; however, after 4 days and to 6 weeks, apoptosis in treated samples was not different than controls. Pro-survival molecules Bcl-2 and Bag-1 were up-regulated upon ER stress in SCs. These results suggest that presence of ECM confers protection from ER stressors. Future studies involving chondrocyte physiology should focus on responses in conditions more closely mimicking the in vivo cartilage environment.
Nugent Ashleigh E; McBurney Denise L; Horton Walter E Jr
Journal of cellular biochemistry
2011
2011-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/jcb.23025" target="_blank" rel="noreferrer noopener">10.1002/jcb.23025</a>
MicroRNA-602 and microRNA-608 regulate sonic hedgehog expression via target sites in the coding region in human chondrocytes.
Academic Medical Centers; Aged; Analysis of Variance; Animal; Animals; Anterior Cruciate Ligament/surgery; Blotting; Cells; Chondrocytes – Physiology; Chondrocytes/drug effects/*metabolism/pathology; Cultured; Disease Models; Female; Funding Source; Gene Expression Regulation/genetics/*physiology; Hedgehog Proteins/genetics/*metabolism; HEK293 Cells; Human; Humans; Immunohistochemistry; In Vitro Techniques; Interleukin-1beta/pharmacology; Knee/etiology/*metabolism/pathology; Male; Matrix Metalloproteinase 13/metabolism; MicroRNAs/genetics/*metabolism; Middle Age; Middle Aged; Ohio; Open Reading Frames/genetics/*physiology; Osteoarthritis; Osteoarthritis – Epidemiology; Osteoarthritis – Physiopathology; Polymerase Chain Reaction; Rabbits; Signal Transduction/genetics/physiology; T-Tests; Transfection; Up-Regulation/drug effects/genetics/physiology; Western
OBJECTIVE: Hedgehog (HH) signaling has recently been associated with cartilage degradation in osteoarthritis (OA). Because interleukin-1beta (IL-1beta) has been implicated as a principal instigator of OA, we sought to determine whether
Akhtar Nahid; Makki Mohammad S; Haqqi Tariq M
Arthritis & rheumatology (Hoboken, N.J.)
2015
2015-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/art.38952" target="_blank" rel="noreferrer noopener">10.1002/art.38952</a>
Fetal lung epithelial ion channels relocate in the cell membrane during late gestation.
Animals; Blotting; Caveolin 1/*metabolism; Cell Membrane/*metabolism; Cl transport; Cystic Fibrosis Transmembrane Conductance Regulator/*metabolism; distal lung fluid absorption; Epithelial Sodium Channels/*metabolism; Epithelium/embryology/*metabolism; Female; fetal lung development; Fetus/*metabolism; Gestational Age; Guinea Pigs; IL-1beta; Immunoprecipitation; Interleukin-1beta/metabolism; Ion Channels/metabolism; Lung/embryology/*metabolism; Na transport; Western
Near the end of gestation, the direction of ion and fluid flow across the alveolar epithelium rapidly changes from secretion to absorption. Thus, the relative cell membrane location of epithelial Na channels (ENaCs) and cystic fibrosis transmembrane regulator (CFTR) Cl channels during late fetal lung development and after maternal interleukin-1beta (IL-1beta) pretreatment was the focus of our study. Western blot analysis after sucrose gradient separation of caveolin-1-(Cav-1)-rich membrane regions (CRR) and Cav-1-poor membrane (non-CRR) revealed primary CRR ENaC localization at gestation day (GD) 61 in guinea pigs. Correlating with the natural induction of distal lung fluid absorption, ENaC appeared in the non-CRR cell membrane regions at GD68. Conversely, CFTR was present in the non-CRR cell membrane regions at GD61 and in the CRRs at GD68.
Beard LaMonta L; Li Tianbo; Hu Yang; Folkesson Hans G
Anatomical record (Hoboken, N.J. : 2007)
2011
2011-09
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/ar.21363" target="_blank" rel="noreferrer noopener">10.1002/ar.21363</a>