New biomarkers are needed to improve knowledge about pathophysiology, in order to provide earlier correct diagnosis and to follow disease progression of the neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and Parkinson's disease (PD). The aim of this thesis was to find new biomarkers for these diseases. First, increased serum levels and unchanged levels in postmortal spinal cord of vascular endothelial growth factor (VEGF) were demonstrated. VEGF was not detected in cerebrospinal fluid (CSF) in ALS. Second, increased levels of fibroblast growth factor 2 were found in the CSF and serum of ALS patients. Both studies used enzyme-linked immunoassays. Third, a proteomics method for CSF analysis was explored, based on tryptic digestion and subsequent separation and detection of the peptides by on-line liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry. ALS-specific patterns were observed. Four out of five samples were correctly assigned, but no single protein biomarker could be identified. Fourth, [11C](L)-deprenyl-D2 (DED) positron emission tomography (PET) demonstrated increased retention in the pons and white matter in ALS. DED binds to monoamino oxidase B, which in the brain is primarily located in astrocytes. Thus evidence was provided that astrocytosis may be detected in vivo in ALS. Fifth, normal [11C]-PIB binding in five nondemented patients with PD was reported, in contrast to previous findings of increased retention in Alzheimer's disease reflecting amyloid aggregation. Finally, the combined use of fluorodeoxyglucose and L-[β 11C]-DOPA PET for the differential diagnosis of parkinsonian syndromes was evaluated. PET provided support for the clinical diagnosis in 62 out of 75 patients, and served to exclude suspected diagnoses in another five patients.

BACKGROUND: Prostate cancer is by far the most common cancer among Swedish men. Some patients have an aggressive lethal disease, but the majority of affected men have long expected survival. Unfortunately, the diagnostic tools available are insufficient in predicting disease aggressiveness. Novel prognostic markers are therefore urgently needed. Furthermore, metastatic prostate cancer is generally treated with castration, but the long-term effects are insufficient. Additional studies are therefore needed to explore how the effects of this therapy can be enhanced. Prostate growth and regression is beside testosterone controlled by locally produced regulators. Vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) are two of the major regulators in the normal prostate and in prostate tumours. MATERIALS AND METHODS: VEGF and EGFR were explored in the prostate, by treating rats with either anti-VEGF or anti-EGFR treatment during castration and testosterone-stimulated prostate growth. Rats with implanted androgen-independent prostate tumours were treated with an inhibitor of both VEGF receptor-2 (VEGFR-2) and EGFR. Stereological techniques, immunohistochemistry, western blotting and quantitative real-time PCR were used to evaluate these experiments. Furthermore, prostate tissue from untreated prostate cancer patients was used to retrospectively explore the expression of phosphorylated-EGFR (pEGFR) in relation to outcome. RESULTS: Anti-VEGF treatment during testosterone-stimulated prostate growth, inhibited vascular and prostate growth. Anti-EGFR treatment during castration and testosterone-stimulated prostate growth resulted in enhanced castration effects and inhibited prostate growth. Anti-vascular treatment of androgen-independent prostate cancer with an inhibitor of VEGFR-2 and EGFR, that targets the normal and tumour vasculature, enhanced the effects of castration. Low immunoreactivity for pEGFR in prostate epithelial cells, both in the tumour and also in the surrounding non-malignant tissue, was associated with good prognosis. CONCLUSIONS: Anti-vascular treatment, with an inhibitor of VEGFR-2 and EGFR, in combination with castration could be an effective way to treat androgen-insensitive prostate tumours. VEGF and EGFR signalling are necessary components in testosterone-stimulated prostate growth. Phosphorylation of EGFR could be a useful prognostic marker for prostate cancer patients. Tumours may affect the surrounding non-malignant tissue and pEGFR immunoreactivity in the morphologically normal prostate tissue can be used to retrieve prognostic information.

This project combines oncolytic Vaccinia virus (VV) with irinotecan (CPT-11) for the treatment of cancer. VV can infect, replicate in and destroy cancer cells, yet leave healthy cells relatively unaffected. CPT-11 is a chemotherapeutic of which ~5% is converted to the more active chemotherapeutic SN-38 by endogenous carboxylesterase (CE) enzymes. SN-38 is a topoisomerase I inhibitor that induces DNA double strand breaks, leading to growth arrest and apoptosis. Consequently, VV has been engineered to express a more effective isoform of the CE enzyme. The virus’ tumour tropism should restrict enhanced conversion of CPT-11 to the tumour. Neither CPT-11 nor SN-38 interfered with VV replication or spread. Engineered recombinants expressed CE enzyme which, when combined with CPT-11, produced DNA double strand breaks and cancer cell death. In vitro, the combination of CE-virus and CPT-11 killed more K-562 cancer cells than its non-CE counterpart and CPT-11.

In sheep, large ovarian antral follicles grow in waves with a periodicity of every 4 to 5 days; each wave is initiated by a peak in serum concentrations of follicle stimulating hormone (FSH). In the present thesis, follicular data and hormone estimations acquired from daily ultrasonography and blood samples, respectively, were used to study mechanisms regulating the number of follicular waves per estrous cycle. Using additional approaches such as implants releasing estradiol-17â and or progesterone, immunization against gonadotropin releasing hormone (GnRH), and injections of GnRH, the role of pulsed luteinizing hormone (LH) secretion and FSH peaks in follicular wave emergence and growth and the dependency of FSH peaks on pulsed GnRH secretion, were studied in sheep. The viability of aged follicles was also addressed.<p> The results of the present studies showed that ewes with three or four waves per cycle had cycles of the same length. The inter-wave interval was longer for the first and the last or ovulatory wave of the cycle in three compared to four wave cycles. The length of the lifespan and regression phase of the largest follicle of a wave declined across the cycle as FSH peak concentration and amplitude decreased. The maximum follicular diameter of the largest follicle growing in the first wave and the last or ovulatory wave of the cycle was greater compared to other waves of the cycle. Treatment of anestrous ewes with estradiol releasing implants alone completely abolished pulsed LH secretion and suppressed follicular wave development; however, FSH secretion was only minimally affected and the pool of small follicles was not affected. When pulsed secretion of LH was restored by frequent injections of GnRH, follicular waves were re-established. Treatment of anestrous ewes with implants releasing estradiol and progesterone, decreased FSH peak amplitude and abolished LH pulses and follicular waves; the size of the pool of small follicles increased. Immunization against GnRH in anestrous ewes abolished pulsatile LH secretion and suppressed follicular wave emergence; however, FSH peaks continued to occur for several weeks. In cyclic ewes, creating an LH pulse frequency typical of the follicular phase, during the luteal phase of the cycle by giving GnRH, increased maximum diameter of the largest follicle in a wave and serum concentrations of estradiol and progesterone. The enhanced growth of follicles in a wave blocked the next expected FSH peak and its associated follicular wave. Decreasing LH pulse frequencies lower than the minimal frequency seen in the luteal phase, by implants releasing progesterone, did not affect the growth of follicular waves.<p> It was previously demonstrated that treatment of non-prolific WWF ewes with Prostaglandin F2á (PGF2á) and medroxy progesterone acetate (MPA) increased the ovulation rate by adding ovulations from the penultimate wave in addition to the final wave of the cycle; however, fertility was not improved. In the last study of my thesis, we collected follicles, with an extended lifespan, from the penultimate wave of the cycle in ewes given the PGF2á and MPA treatment. We compared their quality with follicles from the final wave of the cycle by looking at the expression of markers of follicular development. The results showed that theca cells of follicles from the final wave had significantly higher mRNA expression for vascular endothelial growth factor (VEGF) compared to follicles from the penultimate wave. Granulosa cells of follicles from the final wave had significantly higher mRNA expression for connexion 43 (Cx43) compared to follicles from the penultimate wave. Protein expression for Cx43, proliferating cell nuclear antigen (PCNA) and Factor VIII was greater in follicles from the final compared to the penultimate wave.<p> We concluded from the present studies that: 1) the mechanism that makes a three wave or four wave cycle is unclear; 2) some level of pulsatile LH secretion is required for an FSH peak to trigger emergence of follicular waves in anestrous ewes; 3) progesterone enhances the inhibitory effects of estradiol on FSH secretion in anestrous ewes, suppressing specifically FSH peak amplitude; 4) an endogenous rhythm may exist that drives the peaks in FSH secretion independent of secretory products from the follicles growing in a wave and pulsed GnRH secretion; 5) follicular waves in ewes, when exposed to an LH pulse frequency similar to the follicular phase, during the luteal phase of the cycle, when serum progesterone concentrations are high, can grow and function like ovulatory follicles growing in the follicular phase of the cycle; 6) expression of some markers of vascularization/ angiogenesis, gap-junctional communication and cell proliferation, appeared to be decreased in follicles from the penultimate compared to the final wave of an estrous cycle, when the lifespan of follicles from the penultimate wave was extended such that they were present in the ovary with follicles from the final wave of the cycle.

In sheep, large ovarian antral follicles grow in waves with a periodicity of every 4 to 5 days; each wave is initiated by a peak in serum concentrations of follicle stimulating hormone (FSH). In the present thesis, follicular data and hormone estimations acquired from daily ultrasonography and blood samples, respectively, were used to study mechanisms regulating the number of follicular waves per estrous cycle. Using additional approaches such as implants releasing estradiol-17â and or progesterone, immunization against gonadotropin releasing hormone (GnRH), and injections of GnRH, the role of pulsed luteinizing hormone (LH) secretion and FSH peaks in follicular wave emergence and growth and the dependency of FSH peaks on pulsed GnRH secretion, were studied in sheep. The viability of aged follicles was also addressed.<p> The results of the present studies showed that ewes with three or four waves per cycle had cycles of the same length. The inter-wave interval was longer for the first and the last or ovulatory wave of the cycle in three compared to four wave cycles. The length of the lifespan and regression phase of the largest follicle of a wave declined across the cycle as FSH peak concentration and amplitude decreased. The maximum follicular diameter of the largest follicle growing in the first wave and the last or ovulatory wave of the cycle was greater compared to other waves of the cycle. Treatment of anestrous ewes with estradiol releasing implants alone completely abolished pulsed LH secretion and suppressed follicular wave development; however, FSH secretion was only minimally affected and the pool of small follicles was not affected. When pulsed secretion of LH was restored by frequent injections of GnRH, follicular waves were re-established. Treatment of anestrous ewes with implants releasing estradiol and progesterone, decreased FSH peak amplitude and abolished LH pulses and follicular waves; the size of the pool of small follicles increased. Immunization against GnRH in anestrous ewes abolished pulsatile LH secretion and suppressed follicular wave emergence; however, FSH peaks continued to occur for several weeks. In cyclic ewes, creating an LH pulse frequency typical of the follicular phase, during the luteal phase of the cycle by giving GnRH, increased maximum diameter of the largest follicle in a wave and serum concentrations of estradiol and progesterone. The enhanced growth of follicles in a wave blocked the next expected FSH peak and its associated follicular wave. Decreasing LH pulse frequencies lower than the minimal frequency seen in the luteal phase, by implants releasing progesterone, did not affect the growth of follicular waves.<p> It was previously demonstrated that treatment of non-prolific WWF ewes with Prostaglandin F2á (PGF2á) and medroxy progesterone acetate (MPA) increased the ovulation rate by adding ovulations from the penultimate wave in addition to the final wave of the cycle; however, fertility was not improved. In the last study of my thesis, we collected follicles, with an extended lifespan, from the penultimate wave of the cycle in ewes given the PGF2á and MPA treatment. We compared their quality with follicles from the final wave of the cycle by looking at the expression of markers of follicular development. The results showed that theca cells of follicles from the final wave had significantly higher mRNA expression for vascular endothelial growth factor (VEGF) compared to follicles from the penultimate wave. Granulosa cells of follicles from the final wave had significantly higher mRNA expression for connexion 43 (Cx43) compared to follicles from the penultimate wave. Protein expression for Cx43, proliferating cell nuclear antigen (PCNA) and Factor VIII was greater in follicles from the final compared to the penultimate wave.<p> We concluded from the present studies that: 1) the mechanism that makes a three wave or four wave cycle is unclear; 2) some level of pulsatile LH secretion is required for an FSH peak to trigger emergence of follicular waves in anestrous ewes; 3) progesterone enhances the inhibitory effects of estradiol on FSH secretion in anestrous ewes, suppressing specifically FSH peak amplitude; 4) an endogenous rhythm may exist that drives the peaks in FSH secretion independent of secretory products from the follicles growing in a wave and pulsed GnRH secretion; 5) follicular waves in ewes, when exposed to an LH pulse frequency similar to the follicular phase, during the luteal phase of the cycle, when serum progesterone concentrations are high, can grow and function like ovulatory follicles growing in the follicular phase of the cycle; 6) expression of some markers of vascularization/ angiogenesis, gap-junctional communication and cell proliferation, appeared to be decreased in follicles from the penultimate compared to the final wave of an estrous cycle, when the lifespan of follicles from the penultimate wave was extended such that they were present in the ovary with follicles from the final wave of the cycle.

Im Rahmen dieser Arbeit wurde die Rolle des Rezeptors VEGFR-2, Flk-1, im neuronalen Kontext untersucht. In einem ersten Schritt wurde in embryonalen Stammzellen der Maus das fluoreszierende Protein eGFP unter der Kontrolle regulatorischer Sequenzen des flk-1-Promotors, -Enhancers exprimiert. Nach der Differenzierung zu Sphäroiden wurden Endothelzellen nachgewiesen, die sowohl eGFP als auch das zelltypspezifische Oberflächenantigen CD31 ausprägen. Ebenso wurden nach der neuronalen Differenzierung in Gegenwart von Stromazellen eGFP-exprimierende Zellen identifiziert. Diese standen mit Zellen, die das für neuronale Vorläuferzellen charakteristische Protein Nestin ausprägten, in einem räumlichen Zusammenhang. Die Vorgehensweise, die Inaktivierung des flk-1-Gens mit der Differenzierung embryonaler Stammzellen in vitro zu kombinieren, sollte hier die Interpretation des Phänotyps des flk-1-defizienten Mausmodells ermöglichen. Der Rezeptor war während der neuronalen Differenzierung der Stammzellen auf Stromazellen in vitro für die Regulation der Anzahl der Vorläuferzellen essentiell. Ferner spielte der Rezeptor im Rahmen eines weiteren Differenzierungsmodells, das auf der Zugabe relevanter Wachstumsfaktoren beruht, eine instruktive Rolle im Hinblick auf die Identität der Neuronen. Kriterium war hier die differentielle Expression Homeobox-enthaltender Transkriptionsfaktoren. In einem zweiten Schritt wurden mit Hilfe dieses Modells differentiell-exprimierte Gene von Stammzellen des Wildtyps sowie Zellen mit einer Inaktivierung des flk-1-Gens nach der neuronalen Differenzierung durch subtraktive Hybridisierung in Verbindung mit der PCR identifiziert. Tatsächlich wurde das Protein PEA-15 nicht nur differentiell exprimiert sondern auch als Bestandteil des VEGFR-2-vermittelten Signalwegs identifiziert. Die biologischen Funktionen des Proteins PEA-15 wurden durch VEGF-vermittelte Phosphorylierung reguliert. Die Stimulation durch VEGF führte zunächst zu einer Aktivierung des Proteinkinase B-, Akt-Signalwegs. Für die Stimulation des Akt-Signalwegs war die Phosphorylierung der intrazellulären Tyrosinreste Y1052 und Y1057 des Rezeptors essentiell. Damit einhergehend wurde PEA-15 gegenüber der proteasomalen Degradation stabilisiert. Es wurde gezeigt, daß das Protein PEA-15 die Teilungsaktivität von Zellen beeinflusst. Die VEGF- vermittelte Stimulation führte zur Phosphorylierung der Mitogen-aktivierten Proteinkinasen ERK1 und ERK2. Die weitere Phosphorylierung der Substrate dieser Kinasen im Zellkern wurde durch Interaktion mit PEA-15 unterdrückt. Die Regulation des c-fos-Promotors war zugleich Indikator der Inhibition der Phosphorylierung betreffender Substrate sowie der proliferativen Aktivität. Auf diese Weise ist die Phosphorylierung von PEA-15 nach Stimulation durch VEGF für die Selektivität des Flk-1-vermittelten Signalwegs von unmittelbarer Bedeutung. Die Regulation der biologischen Funktion von PEA-15 erklärt die differentielle Ausprägung im Rahmen der neuronalen Differenzierung embryonaler Stammzellen in vitro. So war die Anzahl GFAP- beziehungsweise PEA-15-exprimierender Zellen nach Differenzierung muriner Stammzellen mit einer Inaktivierung des flk-1-Gens deutlich geringer. Die differentielle Expression identifizierter Gene wurde im Mausmodell nach konditionaler Inaktivierung des flk-1-Gens überprüft. Tatsächlich wurde Vimentin in verschiedenen Arealen des Gehirns differentiell ausgeprägt. Ein Zusammenhang zwischen der differentiellen Expression des Proteins PEA-15, der Anzahl GFAP-exprimierender Zellen und der Ausprägung des Rezeptors Flk-1 ergab sich aus der Identifikation einer Zellpopulation in der subgranulären Zone des Gyrus Dentatus. Dort wurde in flk-1-defizienten, adulten Mäusen eine geringere Anzahl GFAP-exprimierender Zellen nachgewiesen. Schließlich wurden sowohl im Cerebellum als auch im Cortex histologische Unterschiede deutlich, die sich im adulten Organismus aus der Inaktivierung des Rezeptors Flk-1 ergeben. Die vorliegende Arbeit zeigt, daß der Rezeptor VEGFR-2, Flk-1, im neuronalen Kontext eine Rolle spielt, die sich nicht ausschließlich auf die Vermittlung eines Schutzmechanismus gegenüber der neuronalen Apoptose beschränkt, sondern auch auf eine Beteiligung an der Neurogenese hinweist. Die Vorgehensweise, mit Hilfe der subtraktiven Hybridisierung Bestandteile Rezeptor-vermittelter Signalwege vor dem Hintergrund der Differenzierung embryonaler Stammzellen zu identifizieren, verdeutlicht die Eignung der Methode auch bei komplexen Zellpopulationen.

H HARP (heparin-affin regulatory peptide), γνωστή και ως πλειοτροπίνη (PTN), είναι ένας 18 kDa αυξητικός παράγοντας, ο οποίος έχει υψηλή συγγένεια για την ηπαρίνη. Η HARP έχει πολλαπλές βιολογικές δράσεις, όπως συμμετέχει στη ρύθμιση του κυτταρικού πολλαπλασιασμού, στη μετανάστευση και τη διαφοροποίηση. Επιπλέον η έκφραση της σχετίζεται με την φυσιολογική και καρκινική αγγειογένεση in vitro και in vivo. Στην παρούσα εργασία μελετήθηκε η έκφραση της HARP και των υποδοχέων της, ALK και RPTPβ/ζ, στις διάφορες ημέρες ανάπτυξης της CAM εμβρύου όρνιθας. Επίσης, μελετήθηκε η μείωση της έκφρασης της ενδογενούς HARP, με πλασμίδιο που φέρει την αντινοηματική αλληλουχία (AS-HARP), στην αγγειογένεση in vivo, στη φωσφορυλίωση των Εrk1,2 και στη λεμφαγγειογένεση της CAM εμβρύου όρνιθας. Ανάλυση κατά Western και RT-PCR στις διάφορες ημέρες ανάπτυξης του εμβρύου έδειξε ότι η έκφραση της HARP συμβαδίζει με τη δημιουργία νέων αγγείων στη CAM, ενώ η έκφραση των υποδοχέων της HARP στην CAM φαίνεται να είναι αυξημένη στα πρώτα στάδια ανάπτυξης του ιστού. Επίσης, η μείωση της έκφρασης της HARP μετά τη χορήγηση του πλασμιδίου AS-HARP, μείωσε τα επίπεδα της πρωτεΐνης, το μήκος των αγγείων και τη φωσφορυλίωση των Erk1/2 στο in vivo μοντέλο της CAM εμβρύου όρνιθας. Αντίθετα, η μείωση της έκφρασης της HARP μετά τη χορήγηση του πλασμιδίου AS-HARP, δεν επηρέασε τη λεμφαγγειογένεση της CAM εμβρύου όρνιθας. Σαν τελικό συμπέρασμα προκύπτει ότι η έκφραση της ενδογενούς HARP στην CAM εμβρύου όρνιθας είναι σημαντική για τη φυσιολογική αγγειογένεση in vivo. / Heparin-affin regulatory peptide (HARP), also known as pleiotrophin or heparin-binding growth-associated molecule, is an 18 kDa growth factor that has a high affinity for heparin. HARP is involved in the control of cellular proliferation, migration and differentiation. Moreover, there is a strong correlation between HARP expression and tumor growth and angiogenesis. In the present work, we studied the expression of HARP and its receptors, ALK and RPTPβ/ζ, during development of the chicken embryo chorioallantoic membrane (CAM), in relation to angiogenesis. By western blot analysis and RT-PCR, it was shown that HARP, ALK and RPTPβ/ζ expression increased at days of on-going angiogenesis and decreased at later time points. Transfection of CAMs with an anti-sense HARP gene construct led to a significant decrease in HARP amounts compared to vector control transfected CAMs, a significant decrease in the length of CAM blood vessels, and a decrease in the phosphorylation of Erk1/2. Contrary, transfection of CAMs with the anti-sense HARP gene construct had no influence in lymphangiogenesis of the chicken embryo chorioallantoic membrane (CAM). These data suggest that endogenous HARP is involved in angiogenesis in vivo.

This project combines oncolytic Vaccinia virus (VV) with irinotecan (CPT-11) for the treatment of cancer. VV can infect, replicate in and destroy cancer cells, yet leave healthy cells relatively unaffected. CPT-11 is a chemotherapeutic of which ~5% is converted to the more active chemotherapeutic SN-38 by endogenous carboxylesterase (CE) enzymes. SN-38 is a topoisomerase I inhibitor that induces DNA double strand breaks, leading to growth arrest and apoptosis. Consequently, VV has been engineered to express a more effective isoform of the CE enzyme. The virus’ tumour tropism should restrict enhanced conversion of CPT-11 to the tumour. Neither CPT-11 nor SN-38 interfered with VV replication or spread. Engineered recombinants expressed CE enzyme which, when combined with CPT-11, produced DNA double strand breaks and cancer cell death. In vitro, the combination of CE-virus and CPT-11 killed more K-562 cancer cells than its non-CE counterpart and CPT-11.

Both platelet-rich plasma (PRP) and vascular endothelial growth factor (VEGF) can promote regeneration. The aim of this study was to compare the effects of these two elements on bone formation and vascularization in combination with bone marrow stromal cells (BMSC) in a critical-size bone defect in rabbits. The critical-size defects of the radius were filled with: (1) a calcium-deficient hydroxyapatite (CDHA) scaffold + phVEGF165-transfected BMSC (VEGF group), (2) CDHA and PRP, or (3) CDHA, autogenous BMSC, and PRP. As controls served: (4) the CDHA scaffold alone and (5) the CDHA scaffold and autogenous BMSC. The volume of new bone was measured by means of micro-CT scans, and vascularization was assessed in histology after 16 weeks. Bone formation was higher in the PRP + CDHA, BMSC + CDHA, and PRP + BMSC + CDHA groups than in the VEGF group (p < 0.05). VEGF transfection significantly promoted vascularization of the scaffolds in contrast to BMSC and PRP (p < 0.05), but was similar to the result of the CDHA + PRP + BMSC group. The results show that VEGF-transfected BMSC as well as the combination of PRP and BMSC improve vascularization, but bone healing was better with the combination of BMSC and PRP than with VEGF-transfected BMSC. Expression of VEGF in BMSC as a single growth factor does not seem to be as effective for bone formation as expanded BMSC alone or PRP which contains a mixture of growth factors. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

Made available in DSpace on 2016-06-02T19:22:50Z (GMT). No. of bitstreams: 1 2009.pdf: 1332211 bytes, checksum: 8cbbbf935b4f4e81409c0c7f26786e9a (MD5) Previous issue date: 2008-08-11 / Universidade Federal de Sao Carlos / Androgenic-anabolic steroids have been used both for performance improvement and aesthetic reasons. It is well know that high doses of AAS can raise serious adverse effects such as skeletal muscle injuries including increase in the rate of muscle strains/ruptures. The aim of this study was to investigate VEGF mRNA expression in the rat soleus muscle after jumping training associated with androgenic-anabolic steroids (AAS) administration. Wistar rats were grouped into: sedentary (S); trained without AAS (T); nandrolone decanoate (ND)-treated sedentary (AAS); and trained with AAS (AAST). The trained groups have carried out jumps in water at 32°C.: 4 series of 10 jumps each, with a 30-second interval among series, for 7 weeks, with 50- 80% overload of the animal corporal mass. The AAS (Decadurabolin® - 5mg/kg) was injected via subcutaneous in animal back twice a week. Real-time PCR analyses have shown that training significantly increased VEGF mRNA expression in comparison with the S, AAS groups. When training exercise was associated with nandrolone decanoate, the VEGF mRNA expression was inhibit compared with T group. The inhibited expression of VEGF by AAS administration could cause diminished angiogenesis in skeletal muscle. These results suggest that the AAS may be strongly prejudicial to muscle remodeling and performance. / Muitos estudos vêm mostrando os efeitos nocivos para o organismo do abuso de esteróides anabólicos androgênicos (EAA) por atletas e freqüentadores de academia. Entretanto há poucas pesquisas relatando os efeitos prejudiciais dessas substâncias no músculo esquelético. O objetivo desse trabalho foi analisar a expressão do fator de crescimento de endotélio vascular (VEGF) no músculo sóleo de ratos submetidos ao tratamento com EAA e ao exercício de carga. Os animais foram divididos em quatro grupos: S (sedentário controle), T (treinado controle), EAA (sedentário com administração de decanoato de nandrolona) e EAAT (treinado com administração de decanoato de nandrolona). O treinamento foi constituído por saltos em meio líquido à 32°C: 4 séries de 10 saltos cada, com intervalo de 30 segundos entre as séries, em 5 dias da semana, durante 7 semanas, com carga variando de 50 80% da massa corporal do animal. O decanoato de nandrolona (Decadurabolin® - mg/kg) foi injetado via subcutânea no dorso dos animais, duas vezes por semana. A análise da expressão de mRNA de VEGF, por PCR em tempo real mostrou que no grupo que treinou e que não recebeu a injeção de EAA houve aumento significante na expressão desse fator em relação aos grupos que não treinaram (S e EAA). Por outro lado, quando o treino foi associado com a administração de EAA houve inibição da expressão de VEGF em relação ao grupo treinado controle. VEGF é um fator chave na indução da angiogênese, processo de formação de novos vasos sangüíneos a partir de vasos pré-existentes e essa é uma das primeiras adaptações do músculo ao exercício. Em conclusão a diminuição da expressão de VEGF com o uso de EAA poderia provocar uma redução na formação desses novos vasos, promovendo possivelmente uma redução na performance.