JOURNAL OF PHYCOLOGY, Issue 6 2000
Alf Skovgaard
The marine dinoflagellate Gyrodinium resplendens Hulburt is a mixotroph. It possesses chloroplasts and is photosynthetic, and it also feeds phagotrophically on another dinoflagellate, Prorocentrum minimum (Pavillard) Schiller. The species could be cultivated only in food-replete cultures. When kept in cultures without food, cellular chl a content and photosynthetic activity of G. resplendens decreased and growth ceased after a few days. In food-replete cultures, G. resplendens could grow strictly heterotrophically in darkness, but growth rate was then three times lower than in food-replete cultures kept in light. It is suggested that the main importance of phagotrophy is to acquire a growth factor essential to photosynthetic growth. The addition of soil extract or amino acids to the growth medium induced enhanced photosynthetic growth of the species even without the presence of particulate food, but only for approximately 2 weeks. Long-term starvation of G. resplendens led to loss of the ability to feed, and therefore starved cells eventually reached a point of no return where neither photosynthesis nor phagotrophy could sustain further growth. Light microscopical observations on G. resplendens revealed new morphological and behavioral details of the species. [source]

NERVE GROWTH FACTOR RESCUE OF CISPLATIN NEUROTOXICITY IS MEDIATED THROUGH THE HIGH AFFINITY RECEPTOR: STUDIES IN PC12 CELLS AND P75 NULL MOUSE DORSAL ROOT GANGLIA

JOURNAL OF THE PERIPHERAL NERVOUS SYSTEM, Issue 1 2002
SJ Fischer
Nerve growth factor (NGF) rescues dorsal root ganglion neurons and PC12 cells from cisplatin-induced cell death. Two model systems were used to demonstrate that rescue is mediated through the high affinity NGF receptor. In dorsal root ganglion (DRG) neurons isolated from p75(,/,) and control mice, 20 ng/ml NGF completely prevented cisplatin-induced death. In PC12 cells, we overexpressed receptor chimeras between the tumor necrosis factor and NGF receptors. We demonstrated that activation of the intracellular domain of Trk A is responsible for the NGF rescue effect. [source]

TRANSFORMING GROWTH FACTOR-,1 (TGF-,1) GENE EXPRESSION AND ACTIVATION IN THE PATHOGENESIS OF FIBROSIS IN PROTEINURIC RENAL DISEASE IN HUMANS

NEPHROLOGY, Issue 1 2002
Robyn Langham
[source]

PODOCYTE INJURY IS SUPPRESSED BY 1,25-DIHYDROXYVITAMIN D3 VIA MODULATION OF TRANSFORMING GROWTH FACTOR-,1/BONE MORPHOGENETIC PROTEIN-7 SIGNALLING IN PUROMYCIN AMINONUCLEOSIDE NEPHROPATHY RATS

CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 7 2009
Hou-Qin Xiao
SUMMARY 1Accumulating evidence suggests that vitamin D and its analogues are renoprotective. However, the precise mechanisms and the molecular targets by which active vitamin D exerts its beneficial effects remain obscure. The objective of the present study was to evaluate the effect of active vitamin D on rats with puromycin aminonucleoside (PAN) nephropathy, a model that is characterized by predominant podocyte injury. 2The PAN nephropathy rats were created by a single intravenous injection of 100 mg/kg PAN. Changes in renal pathology and podocyte numbers were observed. Real-time polymerase chain reaction (PCR) was performed to examine mRNA expression of nephrin, transforming growth factor (TGF)-,1 and bone morphogenetic protein (BMP)-7. Protein expression of nephrin, TGF-,1, BMP-7 and p-Smad2/3 and p-Smad1/5/8 was examined by immunofluorescence, immunohistochemistry and western blotting, respectively. Rats were treated with 1,25(OH)2D3 by gastric gavage at a dose of 2.5 µg/kg per day, starting 2 days before PAN injection and continuing throughout the experiment. 3A single injection of PAN induced massive proteinuria and elevated serum creatinine on Day 7, both of which were significantly suppressed by 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Immunofluorescence and real-time PCR of the podocyte-associated protein nephrin revealed reduced and discontinuous staining and this change was reversed by 1,25(OH)2D3. In PAN nephropathy rats, TGF-,1 and p-Smad2/3 expression was upregulated, whereas that of BMP-7 and p-Smad1/5/8 was downregulated. Treatment with 1,25(OH)2D3 significantly restored BMP-7/Smad signalling while suppressing TGF-,1/Smad signalling. 4In conclusion, 1,25(OH)2D3 can ameliorate podocyte damage and proteinuria induced by PAN. The beneficial effects of 1,25(OH)2D3 on podocytes may be attributable, in part, to direct modulation of TGF-,1/BMP-7 signalling. [source]

AMINOGUANIDINE AMELIORATES OVEREXPRESSION OF PROSCLEROTIC GROWTH FACTORS AND TYPE IV COLLAGEN DEPOSITION IN EXPERIMENTAL DIABETIC NEPHROPATHY

NEPHROLOGY, Issue 3 2000
Gilbert Re
[source]

RELATIONSHIP BETWEEN MYOFIBROBLAST APOPTOSIS AND GROWTH FACTORS IN THE PATHOGENESIS OF RENAL TUBULOINTERSTITIAL FIBROSIS

NEPHROLOGY, Issue 3 2000
Lane Ac
[source]

Sharp developmental thresholds defined through bistability by antagonistic gradients of retinoic acid and FGF signaling

DEVELOPMENTAL DYNAMICS, Issue 6 2007
Albert Goldbeter
Abstract The establishment of thresholds along morphogen gradients in the embryo is poorly understood. Using mathematical modeling, we show that mutually inhibitory gradients can generate and position sharp morphogen thresholds in the embryonic space. Taking vertebrate segmentation as a paradigm, we demonstrate that the antagonistic gradients of retinoic acid (RA) and Fibroblast Growth Factor (FGF) along the presomitic mesoderm (PSM) may lead to the coexistence of two stable steady states. Here, we propose that this bistability is associated with abrupt switches in the levels of FGF and RA signaling, which permit the synchronized activation of segmentation genes, such as mesp2, in successive cohorts of PSM cells in response to the segmentation clock, thereby defining the future segments. Bistability resulting from mutual inhibition of RA and FGF provides a molecular mechanism for the all-or-none transitions assumed in the "clock and wavefront" somitogenesis model. Given that mutually antagonistic signaling gradients are common in development, such bistable switches could represent an important principle underlying embryonic patterning. Developmental Dynamics 236:1495,1508, 2007. © 2007 Wiley-Liss, Inc. [source]

Nerve growth factor expression in parasympathetic neurons: regulation by sympathetic innervation

EUROPEAN JOURNAL OF NEUROSCIENCE, Issue 12 2000
Wohaib Hasan
Abstract Interactions between sympathetic and parasympathetic nerves are important in regulating visceral target function. Sympathetic nerves are closely apposed to, and form functional synapses with, parasympathetic axons in many effector organs. The molecular mechanisms responsible for these structural and functional interactions are unknown. We explored the possibility that Nerve Growth Factor (NGF) synthesis by parasympathetic neurons provides a mechanism by which sympathetic,parasympathetic interactions are established. Parasympathetic pterygopalatine ganglia NGF-gene expression was examined by in situ hybridization and protein content assessed by immunohistochemistry. Under control conditions, NGF mRNA was present in ,,60% and NGF protein was in 40% of pterygopalatine parasympathetic neurons. Peripheral parasympathetic axons identified by vesicular acetylcholine transporter-immunoreactivity also displayed NGF immunoreactivity. To determine if sympathetic innervation regulates parasympathetic NGF expression, the ipsilateral superior cervical ganglion was excised. Thirty days postsympathectomy, the numbers of NGF mRNA-positive neurons were decreased to 38% and NGF immunoreactive neurons to 15%. This reduction was due to a loss of sympathetic nerve impulse activity, as similar reductions were achieved when superior cervical ganglia were deprived of preganglionic afferent input for 40 days. These findings provide evidence that normally NGF is synthesized by parasympathetic neurons and transported anterogradely to fibre terminals, where it may be available to sympathetic axons. Parasympathetic NGF expression, in turn, is augmented by impulse activity within (and presumably transmitter release from) sympathetic axons. It is suggested that parasympathetic NGF synthesis and its modulation by sympathetic innervation provides a molecular basis for establishment and maintenance of autonomic axo-axonal synaptic interactions. [source]

Epidermal Growth Factor Regulates Amino Acid Transport in Chick Embryo Hepatocytes via Protein Kinase C

EXPERIMENTAL PHYSIOLOGY, Issue 4 2000
Maria Marino
System A-mediated amino acid transport, activation of different steps of signal transduction and involvement of different isoforms of protein kinase C (PKC) have been investigated in chick embryo hepatocytes after epidermal growth factor (EGF) stimulation. EGF rapidly (10 min) increased the rate of aminoisobutyric acid (AIB) uptake in chick embryo hepatocytes freshly isolated on the 19th day of embryonic life, while no change was detectable at other embryonal stages. The growth factor stimulation was abolished by PKC and tyrosine kinase inhibitors and was mimicked by 4-phorbol-12-myristate-13-acetate, dimethyl-2 (PMA). EGF treatment did not modify the phosphorylation of the , isoform of phospholipase C (PLC-,), and inositol trisphosphate (IP3) and intracellular calcium levels, but it induced an increase in PKC activity. Our data show that EGF regulates amino acid uptake, via PKC and without PLC-, activation, only in the last period of chick embryo hepatocyte development. The effects of growth factor on PKC activity suggest the involvement of PKC-, and -, isoforms in EGF modulation of amino acid transport. [source]

The effect of various concentrations of human recombinant epidermal growth factor on split-thickness skin wounds

INTERNATIONAL WOUND JOURNAL, Issue 2 2006
Article first published online: 19 JUN 200
Effet de concentrations variées de facteur de croissance épidermique recombinant sur les plaies cutanées d'épaisseur partielle Le facteur de croissance épidermique (EGF) est un stimulant puissant de l'épithélialisation. Cependant, l'application topique d'EGF pour aider à la réépithélialisation des plaies d'épaisseur partielle reste sujette à controversies. Un total de 10 porcs, chacun soumis à une plaie dd'épaisseur partielle de 4 cm/4 cm ont été traités 2 fois par jour pendant 10 jours pour étudier les effets de l'EGF humain recombinant avec des concentrations de 0,1, 1, 5, 10, 20 ,g/g, l'excipient seul et deux groupes contrôles. Les plaies des groupes contrôles et de l'excipient seul ont obtenu un temps de cicatrisation à 100%(HT100) de 9·31 ± 1·34 et 8·5 ± 1·12 alors que les plaies traitées par la crème à l'EGF en concentration à 0·1 ,g/g (HT100 6·40·71), 1 ,g/g (HT100 5·2 ± 0·63), 5 ,g/g (HT100 5·8 ± 0·85), 10 ug/g (HT100: 7·1 ± 1·45), et 25 ug/g (HT100: 7·4 ± 0·57) ont démontré des reductions de temps d'épithélialisation significatives. Parmi les plaies traitées par EGF, les plaies traitées avec des concentrations de 1 et 5 ,g/g obtenaient les épithélialisations les plus rapides et démontraient une augmentation substantielle d'activité des kératinocytes de la couche basale observée par activié Ki-67. En conclusion, Cette etude démontre l'efficacité du facteur de croissance épidermique humain recombinant dans la réépithélialisation des plaies cutanées d'épaisseur partielle, l'effet étant maximum avec des concentrations en EGF de 1 et 5 ,g/g. Einfluß unterschiedlicher humaner rekombinanter epidermaler Wachstumsfaktoren auf intradermale Wunden Epidermal Growth Factor (EGF) ist ein potentes Stimulans für die Epithelialisierung. Es wird jedoch kontrovers diskutiert, ob die topische Applikation von EGF in interadermalen Wunden eine beschleunigte Wundheilung erzielt. 10 Schweine, die jeweils eine 4 × 4 cm intradermale Wunde erhielten, wurden zweimal täglich mit hr-EGF in den Konzentrationen 0,1, 1, 5, 10 und 25 ug über einen Zeitraum von 10 Tagen behandelt. Eine Wunde wurde lediglich mit dem Vehikel, eine weitere als Kontrollwunde behandelt. Diese beiden Wunden erreichten eine 10% Heilung (HT100) nach 9·31 ± 1·34 and 8·5 ± 1·12 Tagen. Dagegen zeigten die Wunden, die mit 0,1 ug HT100: 6·4 ± 0·71), 1 ug/g (HT100: 5·2 ± 0·63), 5 ug/g (HT100: 5·8 ± 0·85), 10 ug/g (HT100: 7·1 ± 1·45), and 25 ug/g (HT100: 7·4 ± 0·57) eine signifikant verkürzte Heilungsdauer. Innerhalb dieser unteschiedlichen Konzentrationen erwiesen sich die Konzentrationen 1 und 5 ug/g als am effektivsten, was durch die Ki-67 Aktivität gezeigt wurde. Zusammenfassend konnte gezeigt werden, dass der hr-EGF eine effektive Substanz für die Heilung von kutanen Wunden ist mit der höchsten Effektivität bei 1 und 5 ug/g. L'effetto di varie concentrazioni di fattore di crescita umano epidermico ricombinante su ulcere cutanee a spessore parziale Il fattore di crescita epidermico (EGF)è un potente stimolatore della riepitelizzazione. Tuttavia l'applicazione topica di EGF per ottenere una migliore riepitelizzazione in lesioni a spessore parziale è molto controversa. Sono stati trattati un totale di 10 maiali, ognuno con lesioni a spessore parziale di 4 × 4 cm, due volte al giorno per 10 giorni per osservare l'effetto del fattore umano ricombinante EGF in concentrazioni di 0·1, 1, 5, 10, 25 ug/g, del solo veicolo, in due controlli. Le ulcere controllo ed il veicolo hanno mostrato ciascuna un 100% di tempo di guarigione (HT100) di 9·31 ± 1·34 e 8·5 ± 1·12 mentre le lesioni trattate con EGF in unguento a concentrazione di 0·1 ug/g (HT100: 6·4 ± 0·71), 1 ug/g (HT100: 5·2 ± 0·63), 5 ug/g (HT100: 5·8 ± 0·85). 10 ug/g (HT100: 7·1 ± 1·45), e 25 ug/g (HT100: 7·4 ± 0·57) hanno mostrato una significativa riduzione del tempo di guarigione. Tra le lesioni trattate con EGF, le lesioni trattate con concentrazioni di EGF di 1 e 5 ug/g hanno ottenuto la più veloce riepitelizzazione con evidenza di un sostanziale incremento nella attività dei cheratinociti basali osservata attraverso l'attività del Ki-67. In conclusione, questo report dimostra l'efficacia del fattore di crescita umano ricombinante epidermico nel favorire la riepitelizzazione di lesioni a spessore parziale con la migliore applicazione di EGF alle concentrazioni di 1 e 5 ug/g. Efecto de diversas concentraciones del factor de crecimiento epidérmico recombinante humano sobre heridas cutáneas de espesor parcial El factor de crecimiento epidérmico (FCE) es un potente estimulante de la epitelización. No obstante, subsiste un debate sobre la aplicación tópica del FCE para facilitar la reepitelización en heridas de espesor parcial. Se trató a un total de diez cerdos, cada uno con ocho heridas de espesor parcial de 4 × 4 cm, dos veces al día durante 10 días para observar el efecto del FCE recombinante humano a concentraciones de 0,1, 1, 5, 10, 25 ,g/g, el vehículo solo y dos controles. Las heridas de control y las tratadas exclusivamente con el vehículo presentaron un tiempo de curación del 100%(TC100) de 9,31 ± 1,34 y 8,5 ± 1,12, mientras que las heridas tratadas con la pomada de FCE a concentraciones de 0,1 ,g/g (TC100: 6,4 ± 0,71), 1 ,g/g (TC100: 5,2 ± 0,63), 5 ,g/g (TC100: 5,8 ± 0,85), 10 ,g/g (TC100: 7,1 ± 1,45) y 25 ,g/g (TC100: 7,4 ± 0,57) mostraron una reducción significativa del tiempo necesario para conseguir la reepitelización. De las heridas tratadas con FCE, en las que recibieron las concentraciones de 1 y 5 ,g/g se logró una reepitelización más rápida con, signos de incremento considerable de la actividad basal de los queratinocitos observada por medio de la actividad Ki-67. En conclusión, este trabajo demuestra la eficacia del factor de crecimiento epidérmico recombinante humano en facilitar la reepitelización de heridas de espesor parcial, y que la curación más eficiente se obtiene con concentraciones de FCE de 1 y 5 ,g/g. Effekten av varierande koncentrationer av human rekombinant epidermal tillväxtfaktor på delhudssår Epidermala tillväxtfaktorn (EGF)är en potent stimulant av epitelialisering. Topisk applicering av EGF för att befrämja re-epitelialisering av delhudssår har emellertid varit kontroversiellt. Totalt 10 grisar, alla med åtta 4 × 4 cm delhudssår, behandlades två gånger dagligen under 10 dagar för att observera effekten av "human recombinant EGF" i koncentrationer av 0·1, 1, 5, 10, 25 ug/g, vehikel enbart, och två kontroller. Vart och ett av kontroll och vehikel enbart såren uppvisade 100% läkningstid (HT100) på 9·31 ± 1·34 och 8·5 ± 1·12, medan de sår som behandlades med EGF salva med en koncentration av 0·1 ug/g (HT100: 6·4 ± 0·71), 1 ug/g (HT100: 5·2 ± 0·63), 5 ug/g (HT100: 5·8 ± 0·85), 10 ug/g (HT100: 7·1 ± 1·45), och 25 ug/g (HT100: 7·4 ± 0·57) uppvisade signifikant reduktion av tiden för att uppnå re-epitelialisering. Bland de EGF behandlade såren, uppnådde de sår som behandlats med EGF med en koncentration av 1 och 5 ug/g snabbast re-epitelialisering med bevis för märkbar ökning av basal keratinocyte aktivitet iakktagen genom Ki-67 aktivitet. Sammanfattningsvis uppvisar denna rapport den gynnsamma effekten av human rekombinant EGF i re-epitelialisering av delhuds sår. Den gynnsammaste läkningseffekten iakktogs med EGF i koncentrationer av 1 och 5 ug/g. [source]

Hepatocyte Growth Factor Contributes to Fracture Repair by Upregulating the Expression of BMP Receptors,

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 10 2005
Yuuki Imai MD
Abstract Hepatocyte growth factor (HGF) is activated and the expression of BMP receptors (BMPRs) is induced around the fracture site during the early phase of fracture repair. HGF facilitates the expression of BMPRs in mesenchymal cells. This study suggests that HGF contributes to fracture repair by inducing the expression of BMPRs. Introduction: The precise mechanisms that control the upregulation of BMP, BMPRs, and other molecules involved in bone repair are not completely understood. In this study, we hypothesized that HGF, activated through the action of thrombin on the HGF activator, may enhance BMP action through the local induction of BMP or BMPRs. Materials and Methods: Callus samples from tibial fractures in mice were harvested for immunohistochemical analysis of HGF and phosphorylated c-Met, for in situ hybridization of BMPRs, and for real-time RT-PCR analysis for the expression of HGF, c-Met, and BMPRs. To study the changes in gene expression of BMPRs in response to HGF, C3H10T1/2 cells were cultured with or without HGF and harvested for real-time RT-PCR and for Western blot analysis. To evaluate the contribution of HGF to the biological action of BMP2, C3H10T1/2 cells and primary muscle-derived mesenchymal cells were precultured with HGF and cultured with BMP2. In addition, the expression of the luciferase gene linked to the Id1 promoter containing the BMP responsive element and alkaline phosphatase (ALP) activity were assayed. Results: Positive immunostaining of HGF and phosphorylated c-Met was detected around the fracture site at 1 day after the fracture was made. mRNA expression of BMPRs was increased 1 day after fracture and localized in mesenchymal cells at the fracture site. From an in vitro study, the expression of mRNA for BMPRs was elevated by treatment with HGF, but the expression of BMP4 did not change. Western blot analysis also showed the upregulation of BMPR2 by HGF treatment. The results from the luciferase and ALP assays indicated increased responsiveness to BMPs by treating with HGF. Conclusions: This study indicates that HGF is activated and expressed at the fracture site and that HGF induces the upregulation of BMPRs in mesenchymal cells. Furthermore, HGF may facilitate BMP signaling without altering the expression of BMP molecules. [source]

Basic Fibroblast Growth Factor Stimulates Vascular Endothelial Growth Factor Release in Osteoblasts: Divergent Regulation by p42/p44 Mitogen-Activated Protein Kinase and p38 Mitogen-Activated Protein Kinase

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 12 2000
Haruhiko Tokuda
Abstract We previously showed that basic fibroblast growth factor (bFGF) activates p38 mitogen-activated protein (MAP) kinase via Ca2+ mobilization, resulting in interleukin-6 (IL-6) synthesis in osteoblast-like MC3T3-E1 cells. In the present study, we investigated the effect of bFGF on the release of vascular endothelial growth factor (VEGF) in these cells. bFGF stimulated VEGF release dose dependently in the range between 10 and 100 ng/ml. SB203580, an inhibitor of p38 MAP kinase, markedly enhanced the bFGF-induced VEGF release. bFGF induced the phosphorylation of both p42/p44 MAP kinase and p38 MAP kinase. PD98059, an inhibitor of upstream kinase of p42/p44 MAP kinase, reduced the VEGF release. SB203580 enhanced the phosphorylation of p42/p44 MAP kinase induced by bFGF. The enhancement by SB203580 of the bFGF-stimulated VEGF release was suppressed by PD98059. The depletion of extracellular Ca2+ by [ethylenebis-(oxyethylenenitrilo)]tetracetic acid (EGTA) or 1,2-bis-(O -aminophinoxy)-ethane- N,N,N,N -tetracetic acid tetracetoxymethyl ester (BAPTA/AM), a chelator of intracellular Ca2+, suppressed the bFGF-induced VEGF release. A23187, a Ca ionophore, or thapsigargin, known to induce Ca2+ release from intracellular Ca2+ store, stimulated the release of VEGF by itself. A23187 induced the phosphorylation of p42/p44 MAP kinase and p38 MAP kinase. PD98059 suppressed the VEGF release induced by A23187. SB203580 had little effect on either A23187-induced VEGF release or the phosphorylation of p42/p44 MAP kinase by A23187. These results strongly suggest that bFGF stimulates VEGF release through p42/p44 MAP kinase in osteoblasts and that the VEGF release is negatively regulated by bFGF-activated p38 MAP kinase. [source]

Direct and Indirect Actions of Fibroblast Growth Factor 2 on Osteoclastic Bone Resorption in Cultures

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 3 2000
Hiroshi Kawaguchi M.D., Ph.D.
Abstract Fibroblast growth factor 2 (FGF-2 or basic FGF) is known to show variable actions on bone formation and bone resorption. This study was undertaken to elucidate the mechanisms whereby FGF-2 affects bone metabolism, especially bone resorption, using three different culture systems. FGF-2 at 10,9 M and higher concentrations induced osteoclastic cell formation in the coculture system of mouse osteoblastic cells and bone marrow cells, and this induction was abrogated by nonsteroidal anti-inflammatory drugs (NSAIDs). 45Ca release from prelabeled cultured mouse calvariae stimulated by FGF-2 (10,8 M) was also inhibited by NSAIDs, and the inhibition was stronger by NSAIDs, which are more selective for inhibition of cyclooxygenase 2 (COX-2) than COX-1, suggesting the mediation of COX-2 induction. COX-2 was highly expressed and its messenger RNA (mRNA) level was stimulated by FGF-2 in osteoblastic cells whereas it was undetectable or not stimulated by FGF-2 in cells of osteoclast lineage. To further investigate the direct actions of FGF-2 on osteoclasts, resorbed pit formation was compared between cultures of purified osteoclasts and unfractionated bone cells from rabbit long bones. FGF-2 (,10,12 M) stimulated resorbed pit formation by purified osteoclasts with a maximum effect of 2.0-fold at 10,11 M, and no further stimulation was observed at higher concentrations. However, FGF-2 at 10,9 M , 10,8 M stimulated resorbed pit formation by unfractionated bone cells up to 9.7-fold. NS-398, a specific COX-2 inhibitor, did not affect the FGF-2 stimulation on purified osteoclasts but inhibited that on unfractionated bone cells. We conclude that FGF-2 at low concentrations (,10,12 M) acts directly on mature osteoclasts to resorb bone moderately, whereas at high concentrations (,10,9 M) it acts on osteoblastic cells to induce COX-2 and stimulates bone resorption potently. [source]

Role of the Latent Transforming Growth Factor ,,Binding Protein 1 in Fibrillin-Containing Microfibrils in Bone Cells In Vitro and In Vivo

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 1 2000
Sarah L. Dallas
Abstract Latent transforming growth factor ,,binding proteins (LTBPs) are extracellular matrix (ECM) proteins that bind latent transforming growth factor , (TGF-,) and influence its availability in bone and other connective tissues. LTBPs have homology with fibrillins and may have related functions as microfibrillar proteins. However, at present little is known about their structural arrangement in the ECM. By using antibodies against purified LTBP1, against a short peptide in LTBP1, and against epitope-tagged LTBP1 constructs, we have shown colocalization of LTBP1 and fibrillin 1 in microfibrillar structures in the ECM of cultured primary osteoblasts. Immunoelectron microscopy confirmed localization of LTBP1 to 10- to 12-nm microfibrils and suggested an ordered aggregation of LTBP1 into these structures. Early colocalization of LTBP1 with fibronectin suggested a role for fibronectin in the initial assembly of LTBP1 into the matrix; however, in more differentiated osteoblast cultures, LTBP1 and fibronectin 1 were found in distinct fibrillar networks. Overexpression of LTBP1 deletion constructs in osteoblast-like cells showed that N-terminal amino acids 67,467 were sufficient for incorporation into fibrillin-containing microfibrils and suggested that LTBP1 can be produced by cells distant from the site of fibril formation. In embryonic long bones in vivo, LTBP1 and fibrillin 1 colocalized at the surface of newly forming osteoid and bone. However, LTBP1-positive fibrils, which did not contain fibrillin 1, were present in cartilage matrix. These studies show that in addition to regulating TGF,1, LTBP1 may function as a structural component of connective tissue microfibrils. LTBP1 may therefore be a candidate gene for Marfan-related connective tissue disorders in which linkage to fibrillins has been excluded. [source]

Differential cytokine activity and morphology during wound healing in the neonatal and adult rat skin

JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 6 2007
W. Wagner
Abstract Wound-healing mechanisms change during transition from prenatal to postnatal stage. Cytokines are known to play a key role in this process. The current study investigated the differential cytokine activity and healing morphology during healing of incisional skin wounds in rats of the ages neonatal (p0), 3 days old (p3) and adult, after six different healing times (2 hrs to 30 days). All seven tested cytokines (Transforming Growth Factor (TGF) ,, TGF,1, ,,2 and ,,3, IGF 1, Platelet Derived Growth Factor A (PDGF A), basic Fibroblast Growth Factor (bFGF) exhibited higher expression in the adult wounds than at the ages p0 and p3. Expression typically peaked between 12 hrs and 3 days post-wounding, and was not detectable any more at days 10 and 30. The neonate specimen showed more rapid re-epithelialization, far less inflammation and scarring, and larger restitution of original tissue architecture than their adult counterparts, resembling a prenatal healing pattern. The results may encourage the use of neonatal rat skin as a wound-healing model for further studies, instead of the more complicated prenatal animal models. Secondly, the data may recommend inhibition of PDGF A, basic FGF or TGF-,1 as therapeutic targets in efforts to optimize wound healing in the adult organism. [source]

In vitro differentiation of human mesenchymal stem cells to epithelial lineage

JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 3 2007
Virgil P, unescu
Abstract Our study examined whether human bone marrow-derived MSCs are able to differentiate, in vitro, into functional epithelial-like cells. MSCs were isolated from the sternum of 8 patients with different hematological disorders. The surface phenotype of these cells was characterized. To induce epithelial differentiation, MSCs were cultured using Epidermal Growth Factor, Keratinocyte Growth Factor, Hepatocyte Growth Factor and Insulin-like growth Factor-II. Differentiated cells were further characterized both morphologically and functionally by their capacity to express markers with specificity for epithelial lineage. The expression of cytokeratin 19 was assessed by immunocytochemistry, and cytokeratin 18 was evaluated by quantitative RT-PCR (Taq-man). The data demonstrate that human MSCs isolated from human bone marrow can differentiate into epithelial-like cells and may thus serve as a cell source for tissue engineering and cell therapy of epithelial tissue. [source]

Endoglin: An accessory component of the TGF-,-binding receptor-complex with diagnostic, prognostic, and bioimmunotherapeutic potential in human malignancies

JOURNAL OF CELLULAR PHYSIOLOGY, Issue 1 2001
Ester Fonsatti
Endoglin (CD105) is a cell membrane glycoprotein over-expressed on highly proliferating endothelial cells in culture, and on endothelial cells of angiogenetic blood vessels within benign and malignant tissues. CD105 binds several factors of the Transforming Growth Factor (TGF)-, superfamily, and its over-expression modulates cellular responses to TGF-,1. The complex of experimental findings accumulated in the last few years strongly indicate that CD105 is a powerful marker of angiogenesis, and that it might play a critical role in the pathogenesis of vascular diseases and in tumor progression. In this paper, we will review the structural, biological and functional features of CD105, as well as its distribution within normal and neoplastic tissues, emphasizing its foreseeable role as a molecular target for new diagnostic and bioimmunotherapeutic approaches in human malignancies. © 2001 Wiley-Liss, Inc. [source]

Endogenous and Exogenous Fibroblast Growth Factor 2 Support Survival of Chick Retinal Neurons by Control of Neuronal Neuronal bcl-xL and bcl-2 Expression Through a Fibroblast Berowth Factor Receptor 1- and Erk-Dependent Pathway

JOURNAL OF NEUROCHEMISTRY, Issue 1 2000
Laurent Désiré
Abstract : Fibroblast growth factor (FGF) 2 is a survival factor for various cell types, including retinal neurons. However, little is understood about the molecular bases of the neuroprotective role of FGF2 in the retina. In this report, FGF2 survival activity was studied in chick retinal neurons subjected to apoptosis by serum deprivation. Exogenous FGF2 supported neuronal survival after serum deprivation and increased neuronal bcl-xL and bcl-2 expression, through binding to its receptor R1 (FGF-R1), and subsequent extracellular signal-regulated kinase (ERK) activation. Endogenous FGF2 was transiently overexpressed after serum deprivation. Its down-regulation by antisense oligonucleotides and blockade of its signaling pathway (binding to FGF-R1, tyrosine phosphorylation, and ERK inhibition) decreased bcl-xL and bcl-2 levels and and enhanced apoptosis, suggesting that endogenous FGF2 supported neuronal survival through a pathway similar to that of exogenous FGF2. This pathway may serve to up-regulate, or maintain, bcl-xL and bcl-2 levels that normally decrease during the onset of apoptosis. Indeed, long-term ERK activation and high bcl-xL levels are necessary for the survival activity of both exogenous and endogenous FGF2. Because FGF2 is upregulated following retinal injury in vivo, we suggest that an injury-stimulated autocrine/paracrine FGF2 loop may serve to maintain high levels of survival proteins, such as Bcl-xL, through ERK activation in retinal neurons. [source]

Epidermal Growth Factor Induces Oxidative Neuronal Injury in Cortical Culture

JOURNAL OF NEUROCHEMISTRY, Issue 1 2000
Yoo Kyung Cha
Abstract : Recently, we have demonstrated that certain neurotrophic factors can induce oxidative neuronal necrosis by acting at the cognate tyrosine kinase-linked receptors. Epidermal growth factor (EGF) has neurotrophic effects via the tyrosine kinase-linked EGF receptor (EGFR), but its neurotoxic potential has not been studied. Here, we examined this possibility in mouse cortical culture. Exposure of cortical cultures to 1-100 ng/ml EGF induced gradually developing neuronal death, which was complete in 48-72 h ; no injury to astrocytes was noted. Electron microscopic findings of EGF-induced neuronal death were consistent with necrosis ; severe mitochondrial swelling and disruption of cytoplasmic membrane occurred, whereas nuclei appeared relatively intact. The EGF-induced neuronal death was accompanied by increased free radical generation and blocked by the anti-oxidant Trolox. Suggesting mediation by the EGFR, an EGFR tyrosine kinase-specific inhibitor, C56, attenuated EGF-induced neuronal death. In addition, inhibitors of extracellular signal-regulated protein kinase 1/2 (Erk-1/2) (PD98056), protein kinase A (H89), and protein kinase C (GF109203X) blocked EGF-induced neuronal death. A p38 mitogen-activated protein kinase inhibitor (SB203580) or glutamate antagonists (MK-801 and 6-cyano-7-nitroquinoxaline-2,3-dione) showed no protective effect. The present results suggest that prolonged activation of the EGFR may trigger oxidative neuronal injury in central neurons. [source]

p38 MAPK inhibition modulates rabbit nucleus pulposus cell response to IL-1,

JOURNAL OF ORTHOPAEDIC RESEARCH, Issue 7 2008
Rebecca K. Studer
Abstract Analysis of disc gene expression implicated IL-1 in the development of intervertebral disc degeneration (IDD) in a rabbit stab model. The purpose of these studies is to determine the role of p38 Mitogen Activated Protein Kinase (p38 MAPK) signaling in nucleus pulposus cell response to IL-1, and to compare rabbit nucleus pulposus (rNP) cell responses to IL-1 activation with those in a stab model of disc degeneration. NP cells maintained in alginate bead culture were exposed to IL-1, with or without p38 MAPK inhibition. RNA was isolated for reverse transcription polymerase chain reaction (RT-PCR) analysis of gene expression, conditioned media analyzed for accumulation of nitric oxide (NO) and prostaglandin E-2 (PGE-2), and proteoglycan synthesis measured after 10 days. IL-1 upregulation of mRNA for cycloxygenase-2 (COX-2), matrix metalloproteinase-3 (MMP-3), IL-1, and IL-6, was blunted by p38 inhibition while downregulation of matrix proteins (collagen I, collagen II, aggrecan) and insulin-like-growth-factor I (IFG-1) was also reversed. mRNA for tissue inhibitor of matrixmetalloproteinase-1 (TIMP-1) was modestly increased by IL-1, while those for Transforming Growth Factor-, (TGF-,) SOX-9, and versican remained unchanged. Blocking p38 MAPK reduced IL-1 induced NO and PGE-2 accumulation and partially restored proteoglycan synthesis. p38 MAPK inhibition in control cells increased mRNA for matrix proteins (aggrecan, collagen II, versican, collagen I) and anabolic factors (IGF-1, TGF, and SOX-9) from 50% to 120%, decreased basal PGE-2 accumulation, but had no effect on message for TIMP-1, MMP-3, or COX-2. Inhibition of p38 MAPK in cytokine-activated disc cells blunts gene expression and production of factors associated with inflammation, pain, and disc matrix catabolism while reversing IL-1 downregulation of matrix protein gene expression and proteoglycan synthesis. The results support the hypothesis that IL-1 could be responsible for many of the mRNA changes seen in rabbit NP in the stab model of disc degeneration, and uphold the concept that development of molecular techniques to block p38 MAPK could provide a therapeutic approach to slow the course of intervertebral disc degeneration. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:991,998, 2008 [source]

Phosphatidylethanol Mediates its Effects on the Vascular Endothelial Growth Factor via HDL Receptor in Endothelial Cells

ALCOHOLISM, Issue 2 2009
Marja Katriina Liisanantti
Background:, Previous epidemiological studies have shown that light to moderate alcohol consumption has protective effects against coronary heart disease but the mechanisms of the beneficial effect of alcohol are not known. Ethanol may increase high density lipoprotein (HDL) cholesterol concentration, augment the reverse cholesterol transport, or regulate growth factors or adhesion molecules. To study whether qualitative changes in HDL phospholipids mediate part of the beneficial effects of alcohol on atherosclerosis by HDL receptor, we investigated whether phosphatidylethanol (PEth) in HDL particles affects the secretion of vascular endothelial growth factor (VEGF) by a human scavenger receptor CD36 and LIMPII analog-I (CLA-1)-mediated pathway. Methods:, Human EA.hy 926 endothelial cells were incubated in the presence of native HDL or PEth-HDL. VEGF concentration and CLA-1 protein expression were measured. Human CLA-1 receptor-mediated mechanisms in endothelial cells were studied using CLA-1 blocking antibody and protein kinase inhibitors. Results:, Phosphatidylethanol-containing HDL particles caused a 6-fold increase in the expression of CLA-1 in endothelial cells compared with the effect of native HDL. That emergent effect was mediated mainly through protein kinase C and p44/42 mitogen-activated protein kinase pathways. PEth increased the secretion of VEGF and that increase could be abolished by a CLA-1 blocking antibody. Conclusions:, High density lipoprotein particles containing PEth bind to CLA-1 receptor and thereby increase the secretion of VEGF from endothelial cells. Ethanol-induced protective effects against coronary heart disease may be explained, at least partly, by the effects of PEth-modified HDL particles on VEGF via CLA-1-mediated mechanisms in endothelial cells. [source]

Effects of Ethanol and Transforming Growth Factor , (TGF,) on Neuronal Proliferation and nCAM Expression

ALCOHOLISM, Issue 8 2002
Michael W. Miller
Background Developmental events targeted by ethanol are cell proliferation, neuronal migration, and neurite outgrowth; the latter processes being mediated by neural cell adhesion molecule (nCAM). TGF,1 affects all three of these events. Therefore, the effects of ethanol on transforming growth factor (TGF) ,1 mediated activities in neocortical neurons in vitro were examined. Methods Primary cultures of cortical neurons were obtained from 16-day-old fetuses and were treated with TGF,1 (0 or 10 ng/ml) and ethanol (0 or 400 mg/dl) for 48 hr. The effects of these substances on cell numbers, [3H]thymidine incorporation, and the expression of nCAM were determined. Results Both cell growth (the change in cell numbers over time) and cell proliferation were inhibited by TGF,1 and ethanol. The action of these two anti-mitogenic factors was additive. In contrast, TGF,1 also promoted the expression of three isoforms of nCAM. Likewise, ethanol also up-regulated nCAM expression. On the other hand, ethanol blocked TGF,1-mediated nCAM expression, particularly of the 120 and 180 kDa isoforms. Conclusions TGF, ligands inhibit neuronal proliferation and stimulate the expression of cell adhesion proteins that promote the movement of postmitotic neurons and process outgrowth. Ethanol alters these phenomena as well. Thus, in neurons, as in astrocytes, TGF,1 and ethanol may interact. [source]

Novel Biomarkers for Diagnosis and Therapeutic Assessment of Overactive Bladder: Urinary Nerve Growth Factor and Detrusor Wall Thickness

LUTS, Issue 2009
Hann-Chorng KUO
Clinical diagnosis of overactive bladder (OAB) varies greatly and is based on subjective symptoms. A better way to diagnose and assess therapeutic outcome in patients who present with OAB needs to be developed. Evidence has shown that urinary proteins, such as nerve growth factor (NGF) and prostaglandin E2 (PGE2) levels increase in patients with OAB, bladder outlet obstruction (BOO) and detrusor overactivity (DO). Urinary NGF level increases physiologically in normal subjects at urge to void, but increases pathologically in OAB patients at small bladder volume and at urgency sensation. Patients with OAB dry and OAB wet have significantly higher urinary NGF levels compared to controls and patients with increased bladder sensation. Urinary NGF levels decrease after antimuscarinic therapy and further decrease after detrusor botulinum toxin injections in refractory OAB. A higher urinary NGF level could be a biomarker for sensory nerve-mediated DO. Urinary NGF levels could be a potential biomarker for diagnosis of OAB and serve for the assessment of the therapeutic effect of antimuscarinic therapy. Another potential biomarker for the diagnosis of OAB is detrusor wall thickness. It has been hypothesized that the bladder wall increases in thickness in patients with OAB. The thickened detrusor wall might decrease in response to antimuscarinic treatment, and measurement of detrusor wall thickness might be a useful biomarker for the evaluation of OAB. However, current investigations do not yet provide a uniform observation among various studies. [source]

High resolution structure of the HDGF PWWP domain: A potential DNA binding domain

PROTEIN SCIENCE, Issue 2 2006
Stephen M. Lukasik
Abstract Hepatoma Derived Growth Factor (HDGF) is an endogenous nuclear-targeted mitogen that is linked with human disease. HDGF is a member of the weakly conserved PWWP domain family. This 70,amino acid motif, originally identified from the WHSC1 gene, has been found in more than 60 eukaryotic proteins. In addition to the PWWP domain, many proteins in this class contain known chromatin remodeling domains, suggesting a role for HDGF in chromatin remodeling. We have determined the NMR structure of the HDGF PWWP domain to high resolution using a combination of NOEs, J-couplings, and dipolar couplings. Comparison of this structure to a previously determined structure of the HDGF PWWP domain shows a significant difference in the C-terminal region. Comparison to structures of other PWWP domains shows a high degree of similarity to the PWWP domain structures from Dnmt3b and mHRP. The results of selected and amplified binding assay and NMR titrations with DNA suggest that the HDGF PWWP domain may function as a nonspecific DNA-binding domain. Based on the NMR titrations, we propose a model of the interaction of the PWWP domain with DNA. [source]

Ablation of Systemic Phosphate-Regulating Gene Fibroblast Growth Factor 23 (Fgf23) Compromises the Dentoalveolar Complex

THE ANATOMICAL RECORD : ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY, Issue 7 2010
E.Y. Chu
Abstract Fibroblast growth factor-23 (FGF23) is a hormone that modulates circulating phosphate (Pi) levels by controlling Pi reabsorption from the kidneys. When FGF23 levels are deficient, as in tumoral calcinosis patients, hyperphosphatemia ensues. We show here in a murine model that Fgf23 ablation disrupted morphology and protein expression within the dentoalveolar complex. Ectopic matrix formation in pulp chambers, odontoblast layer disruption, narrowing of periodontal ligament space, and alteration of cementum structure were observed in histological and electron microscopy sections. Because serum Pi levels are dramatically elevated in Fgf23,/,, we assayed for apoptosis and expression of members from the small integrin-binding ligand, N-linked glycoprotein (SIBLING) family, both of which are sensitive to elevated Piin vitro. Unlike X-linked hypophosphatemic (Hyp) and wild-type (WT) specimens, numerous apoptotic osteocytes and osteoblasts were detected in Fgf23,/, specimens. Further, in comparison to Hyp and WT samples, decreased bone sialoprotein and elevated dentin matrix protein-1 protein levels were observed in cementum of Fgf23,/, mice. Additional dentin-associated proteins, such as dentin sialoprotein and dentin phosphoprotein, exhibited altered localization in both Fgf23,/, and Hyp samples. Based on these results, we propose that FGF23 and (Pi) homeostasis play a significant role in maintenance of the dentoalveolar complex. Anat Rec 293:1214,1226, 2010. © 2010 Wiley-Liss, Inc. [source]

ORIGINAL ARTICLE: Keratinocyte Growth Factor Stimulates Macrophage Inflammatory Protein 3, and Keratinocyte-derived Chemokine Secretion by Mouse Uterine Epithelial Cells

AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 3 2010
Severina N. Haddad
Citation Haddad SN, Wira CR. Keratinocyte growth factor stimulates macrophage inflammatory protein 3, and keratinocyte-derived chemokine secretion by mouse uterine epithelial cells. Am J Reprod Immunol 2010; 64: 197,211 Problem, Communication between uterine epithelial cells and the underlying stromal fibroblasts is critical for proper endometrial function. Stromal fibroblast-derived growth factors have been shown to regulate epithelial immune functions. The purpose of this study was to determine whether keratinocyte growth factor (KGF) regulates uterine epithelial cell chemokine and antimicrobial secretion. Method of study, Uterine epithelial cells were isolated from Balb/c mice and cultured in either 96-well plates or transwell inserts. Epithelial cells were treated with KGF, epidermal growth factor (EGF), or hepatocyte growth factor (HGF). Macrophage inflammatory protein 3, (MIP3,) and keratinocyte-derived chemokine (KC) levels were measured by ELISA. Results, Keratinocyte growth factor stimulated the secretion of MIP3, and KC. The effects on MIP3, by KGF were specific because EGF and HGF had no effect. In contrast, KGF, EGF, and HGF had similar effects on KC. Furthermore, KGF administered to the apical side of epithelial cells had no effect on MIP3, or KC secretion, indicating that the KGF receptor is located on the basolateral surface of uterine epithelial cells. Conclusion, We demonstrate that KGF plays a role in uterine epithelial cell secretion of MIP3, and KC, key immune mediators involved in the protection of mucosal surfaces in the female reproductive tract. [source]

ORIGINAL ARTICLE: Association Study of Vascular Endothelial Growth Factor and Polymorphisms of its Gene with Ectopic Pregnancy

AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 2 2010
Julio Elito Jr
Citation Elito J Jr, Daher S, Fernandes da Silva MO, Marconi NMH, Pendeloski KPT, Moron AF, Camano L. Association study of vascular endothelial growth factor and polymorphisms of its gene with ectopic pregnancy. Am J Reprod Immunol 2010; 63: 120,125 Problem, In ectopic pregnancy, increased levels of vascular endothelial growth factor are present. The aims of this study were to determine the association between ,634C/G, ,460T/C, and +936C/T vascular endothelial growth factor (VEGF) polymorphisms and ectopic pregnancy, and to determine whether serum levels of VEGF were affected by genetic factors. Method of study, This is a case,control study wherein 74 women with a history of ectopic pregnancy in a tertiary care center were compared to 134 post-menopausal controls with two pregnancies and no ectopic pregnancy for the genotyping of VEGF polymorphisms. For 35 patients with the diagnosis of ectopic pregnancy, serum concentrations of VEGF were obtained before the treatment. Genotyping of VEGF (,634C/G, ,460T/C, and +936C/T) polymorphisms was performed by PCR, followed by endonuclease digestion. ELISA was performed to evaluate the VEGF serum levels. Results, The ,634C/G, ,460T/C, and +936C/T VEGF polymorphisms were not associated with ectopic pregnancy (P = 0.170, P = 0.285, and P = 0.700, respectively). The serum levels of VEGF were not associated with the genotype of ,634C/G, ,460T/C, and +936C/T VEGF polymorphisms (P = 0.702; P = 0.347, and P = 0.256, respectively). Conclusion, There was no association between ectopic pregnancy and ,634C/G, ,460T/C, and +936C/T VEGF polymorphisms. There was no correlation between VEGF genotype and the expression of VEGF in blood samples. [source]

Fibroblast Growth Factor 2 Promotes Endothelial Differentiation of Adipose Tissue-Derived Stem Cells

THE JOURNAL OF SEXUAL MEDICINE, Issue 4 2009
Hongxiu Ning PhD
ABSTRACT Introduction., Adipose tissue-derived stem cells (ADSC) could potentially restore endothelial function in vasculogenic erectile dysfunction (ED). The mechanism for ADSC endothelial differentiation remained unidentified. Aim., To test whether ADSC could differentiate into endothelial cells in the penis and to identify the underlying mechanism of ADSC endothelial differentiation. Methods., For in vivo endothelial differentiation, ADSC were labeled with bromodeoxyuridine (BrdU), injected into rat corpora cavernosa, and localized by immunofluorescence and phase-contrast microscopy. For in vitro endothelial differentiation, ADSC were grown in endothelial growth medium 2 (EGM2), stained for endothelial markers CD31, von Willebrand Factor (vWF), and endothelial nitric oxide synthase (eNOS), and assessed for the ability to form tube-like structures in Matrigel and to endocytose acetylated low-density lipoprotein (Ac-LDL). To identify factors that promote ADSC endothelial differentiation, ADSC were grown in various media, each of which contained a specific combination of supplemental factors and assessed for LDL-uptake. PD173074, a selective inhibitor of fibroblast growth factor 2 (FGF2) receptor, was used to confirm the importance of FGF2 signaling for ADSC endothelial differentiation. Main Outcome Measures., In vivo endothelial differentiation was assessed by immunofluorescence microscopy. In vitro endothelial differentiation was assessed by immunofluorescence, Matrigel tube formation, and Ac-LDL uptake. Results., Injected ADSC were localized to the sinusoid endothelium, some of which stained positive for both BrdU and endothelial antigen rat endothelial cell antigen. ADSC proliferated at a faster rate in EGM2 than in standard DMEM, expressed endothelial markers CD31, vWF, and eNOS, formed tube-like structures in Matrigel, and endocytosed Ac-LDL. These properties were greatly diminished when ADSC were grown in the absence of FGF2 but were unaffected when grown in the absence of vascular endothelial growth factor, insulin-like growth factor, or epidermal growth factor. Furthermore, ADSC displayed similar endothelial properties when grown in FGF2-supplemented basic medium as in EGM2. Finally, blockade of FGF2 signaling with PD173074 abrogated ADSC endothelial differentiation. Conclusions., ADSC could differentiate into endothelial cells in the penis. FGF2 signaling mediates ADSC endothelial differentiation. Ning H, Liu G, Lin G, Yang R, Lue TF, and Lin CS. FGF2 promotes endothelial differentiation of adipose tissue-derived stem cells. J Sex Med **;**:**,**. [source]

Role of Increased Penile Expression of Transforming Growth Factor-,1 and Activation of the Smad Signaling Pathway in Erectile Dysfunction in Streptozotocin-Induced Diabetic Rats

THE JOURNAL OF SEXUAL MEDICINE, Issue 10 2008
Lu Wei Zhang MD
ABSTRACT Introduction., It has been suggested that transforming growth factor-,1 (TGF-,1) plays an important role in the pathogenesis of diabetes-induced erectile dysfunction. Aim., To investigate the expression and activity of Smad transcriptional factors, the key molecules for the initiation of TGF-,-mediated fibrosis, in the penis of streptozotocin (STZ)-induced diabetic rats. Methods., Fifty-two 8-week-old Sprague,Dawley rats were used and divided into control and diabetic groups. Diabetes was induced by an intravenous injection of STZ. Main Outcome Measures., Eight weeks later, erectile function was measured by electrical stimulation of the cavernous nerve (N = 12 per group). The penis was harvested and stained with Masson trichrome or antibody to TGF-,1, phospho-Smad2 (P-Smad2), smooth muscle ,-actin, and factor VIII (N = 12 per group). Penis specimens from a separate group of animals were used for TGF-,1 enzyme-linked immunosorbent assay (ELISA), P-Smad2/Smad2, phospho-Smad3 (P-Smad3)/Smad3, fibronectin, collagen I, and collagen IV western blot, or hydroxyproline determination. Results., Erectile function was significantly reduced in diabetic rats compared with that in controls. The expression of TGF-,1, P-Smad2, and P-Smad3 protein evaluated by ELISA or western blot was higher in diabetic rats than in controls. Compared with that in control rats, P-Smad2 expression was higher mainly in smooth muscle cells and fibroblasts of diabetic rats, whereas no significant differences were noted in endothelial cells or in the dorsal nerve bundle. Cavernous smooth muscle and endothelial cell contents were lower in diabetic rats than in controls. Cavernous fibronectin, collagen IV, and hydroxyproline content was significantly higher in diabetic rats than in controls. Conclusion., Upregulation of TGF-,1 and activation of the Smad signaling pathway in the penis of diabetic rats might play important roles in diabetes-induced structural changes and deterioration of erectile function. Zhang LW, Piao S, Choi MJ, Shin H-Y, Jin H-R, Kim WJ, Song SU, Han J-Y, Park SH, Mamura M, Kim S-J, Ryu J-K, and Suh J-K. Role of increased penile expression of transforming growth factor-,1 and activation of the Smad signaling pathway in erectile dysfunction in streptozotocin-induced diabetic rats. J Sex Med 2008;5:2318,2329. [source]

REVIEW ARTICLE: Immunopathogenesis of Pelvic Endometriosis: Role of Hepatocyte Growth Factor, Macrophages and Ovarian Steroids

AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 5 2008
Khaleque Newaz Khan
Endometriosis, a chronic disease characterized by endometrial tissue located outside the uterine cavity is associated with chronic pelvic pain and infertility. However, an in-depth understanding of the pathophysiology of endometriosis is still elusive. It is generally believed that besides ovarian steroid hormones, the growth of endometriosis can be regulated by innate immune system in pelvic microenvironment by their interaction with endometrial cells and immune cells. We conducted a series of studies in perspectives of pelvic inflammation that is triggered primarily by bacterial endotoxin (lipopolysacccharide) and is mediated by toll-like receptor 4 and showed their involvement in the development of pelvic endometriosis. As a cellular component of innate immune system, macrophages were found to play a central role in inducing pelvic inflammatory reaction. We further report here that peritoneal macrophages retain receptors encoding for estrogen and progesterone and ovarian steroids also participate in producing an inflammatory response in pelvic cavity and are involved in the growth of endometriosis either alone or in combination with hepatocyte growth factor (HGF). As a pleiotropic growth factor, HGF retains multifunctional role in endometriosis. We describe here the individual and step-wise role of HGF, macrophages and ovarian steroid hormones and their orchestrated involvement in the immunopathogenesis of pelvic endometriosis. [source]