Chondroitin Sulfate Protects the Liver in an Experimental Model of Extra-Hepatic Cholestasis Induced by Common Bile Duct Ligation.

During liver fibrogenesis, there is an imbalance between regeneration and wound healing. The current treatment is the withdrawal of the causing agent; thus, investigation of new and effective treatments is important. Studies have highlighted the action of chondroitin sulfate (CS) in different cells; thus, our aim was to analyze its effect on an experimental model of bile duct ligation (BDL). Adult Wistar rats were subjected to BDL and treated with CS for 7, 14, 21, or 28 days intraperitoneally. We performed histomorphometric analyses on Picrosirius-stained liver sections. Cell death was analyzed according to caspase-3 and cathepsin B activity and using a TUNEL assay. Regeneration was evaluated using PCNA immunohistochemistry. BDL led to increased collagen content with corresponding decreased liver parenchyma. CS treatment reduced total collagen and increased parenchyma content after 21 and 28 days. The treatment also promoted changes in the hepatic collagen type III/I ratio. Furthermore, it was observed that CS treatment reduced caspase-3 activity and the percentage of TUNEL-positive cells after 14 days and cathepsin B activity only after 28 days. The regeneration increased after 14, 21, and 28 days of CS treatment. In conclusion, our study showed a promising hepatoprotective action of CS in fibrogenesis induced by BDL.

Electrotherapy for urethral modulation: Are extracellular matrix molecules and growth factors potential targets?

AIMS: To evaluate the expression of genes and proteins involved in the urethral components: vessels, nerves, and extracellular matrix, in female rats after trauma by vaginal distension (VD) and after electrical stimulation therapy (electrotherapy). METHODS: We analyzed the urethras of three groups of 18 female rats 30 days posttrauma by VD: control (no interventions); trauma (animals that had VD); and electrotherapy group (those that had VD and were treated with electrical stimulation). We compared the expression of vascular endothelial growth factor (VEGF), nerve growth factor (NGF), collagen types I and III (COL1a1 and COL3a1), and lysyl-oxidase like 1 (LOXL1) among the groups. Real-time reverse transcription-polymerase chain reaction, Western blot, and immunohistochemistry were used for molecule quantification. We used the Kruskal-Wallis test and analysis of variance for statistical analyses with p < 0.05 for significance. RESULTS: The COL1a1 gene expression was higher in the electrotherapy group than the trauma group (p = 0.036). COL3a1, VEGF, NGF, LOXL1 messenger RNA (mRNA) expression did not differ among the groups (p ≥ 0.05). COL1a1, COL3a1, VEGF, NGF, LOXL1 protein levels did not significantly differ among the groups (p ≥ 0.05) in Western blot analysis or immunohistochemistry assays. CONCLUSIONS: Electrotherapy caused a long-term increase in the COL1a1 mRNA level but did not change COL1a1 protein expression or VEGF, NGF, COL3a1, and LOXL1 genes and proteins in the urethras of rats after trauma by VD.

The molecular effects of electrical stimulation on the muscle components of the urethra of female rats after trauma by vaginal distention.

AIMS: To evaluate the expression of genes and proteins related to the urethral muscles of female rats after trauma by vaginal distention (VD) and after electrical stimulation therapy (EST). METHODS: We compared the urethras of four groups of 20 animals each: control without trauma (C), 7 (recent-trauma) and 30 days (late-trauma) post-VD, and VD-treated with EST. We evaluated the expression of myogenic regulatory factors MYOD1 and myogenin (MYOG); skeletal muscle myosin heavy chain 1, 2, and 3 (MYH1, MYH2, and MYH3); smooth muscle MYH11; and myosin light chain 9 (MYL9). We used real-time quantitative polymerase chain reaction, Western blot analysis, and immunohistochemistry. RESULTS: MYOD1 and MYOG genes were overexpressed in the recent-trauma group compared with the other groups (P < .05). MYH1 and MYH3 genes were upregulated in the recent-trauma group compared with the control and EST groups (P < .05). The MYH2 gene was overexpressed in the late-trauma group (P < .05), while the MYH2 protein was significantly increased in the EST group compared with control, recent-trauma and late-trauma groups by 5-, 3-, and 2.7-fold change, respectively (P < .05). MYL9 and MYH11 messenger RNA were overexpressed in both trauma groups compared with control and EST groups (P < .05). MYH11 protein was not different among the study groups (P > .05). CONCLUSIONS: EST enhances the recovery of the damaged urethral tissue of rats mainly by acting on the striated-muscle components. The MYH2 pathway underlies the positive effects of EST in the external urethral sphincter.

Molecular and histomorphological evaluation of female rats' urethral tissues after an innovative trauma model of prolonged vaginal distention: immediate, short-term and long-term effects.

INTRODUCTION AND HYPOTHESIS: An animal model of vaginal distention (VD) was developed to reproduce the acute urethral injury and deficiency underlying stress urinary incontinence (SUI). Data on the chronic effects of urethral trauma and the recovery process are still scarce. We investigated acute, short- and long-term histomorphological and molecular changes in the urethra of rats post 12-h intermittent VD. METHODS: We evaluated the urethra of four groups of female rats (n = 72): control without trauma, 1 h, 7 days and 30 days post VD. We compared the gene and protein expression of the VEGF and NGF growth factors, collagens (COL1a1 and COL3a1), desmin, smooth muscle myosin (MYH11), skeletal muscle myosins (MYH1, MYH2 and MYH3) and cell proliferation marker MKi67. We used real-time RT-qPCR, and immunohistochemistry. RESULTS: Histology showed urethral damage after VD mainly involving the muscular layers. VEGF, NGF, desmin and MKi67 mRNA were significantly upregulated in the urethras of rats 1-h post VD compared with controls (P < 0.05 for all). By 7 days post trauma, COL1a1, MYH11 and MYH3 genes were overexpressed compared with controls (p < 0.05 for all). The COL3a1 protein level was increased by 2.6 times by day 7, while MYH2 protein was significantly decreased (around two times) from 7 to 30 days post VD compared with controls (p < 0.05 for both). CONCLUSIONS: The 12-h intermittent VD causes chronic alterations in the urethra represented by increased COL3a1 and decreased MYH2 protein levels in the long term. The model can potentially be used to study the mechanisms of urethral injury and recovery as well as the physiopathology of SUI.

Molecular and immunohistochemical analysis of the urethra of female rats after induced trauma and intravenous therapy with muscle derived stem cells.

AIMS: To identify the urethral migration of muscle derived stem cells (MDSCs) after intravenous (IV) injection in rats that underwent vaginal distension (VD) and to analyze the effects of MDSC in the urethra of rats after trauma in regards to: (1) mRNA expression of collagens, Vegf, Ngf, Ki67, Myh11, and Myh2; (2) expression of smooth and striated muscle proteins. METHODS: MDSCs expressing green fluorescent protein (GFP) were injected into the tail vein of rats 3 days after VD. The location of GFP cells was verified at 2 h and at 7 days following IV injection. Urethras of three groups were analyzed: Control, Trauma 7D, and MDSC 7D. Real-time RT-qPCR and immunohistochemistry were performed. RESULTS: MDSCs were identified only after 2 h of the procedure in the urethra. Myh11 gene was overexpressed in the Trauma group in relation to Control. Ki67 gene expression was increased in the MDSC group relative to Trauma and Control. Col1a1 and Col3a1 genes expression were increased in the MDSC group relative to Control. Ngf mRNA level was decreased in the MDSC group in relation to Trauma. Protein expression of Mhy11, Myh2, and Desmin were increased in the MDSC group in relation to Trauma and decreased in the Trauma in relation to Control. CONCLUSION: MDSCs migrated early to the traumatized urethra, but did not integrate into the tissue. MDSC alters the expression of genes related to cell proliferation, neural growth factor and extracellular matrix and the expression of smooth and striated muscle proteins in the traumatized rat urethra.

Special dyeing, histochemistry, immunohistochemistry and ultrastructure: A study of mast cells/eosinophilic granules cells (MCs/EGC) from Centropomus parallelus intestine.

Intestine mast cells/eosinophilic granule cells (MCs/EGC) of the marine species Centropomus parallelus (fat snook) were first studied using light and electron microscopy techniques. Mast cells are cells from the connective tissue found in almost all organs and tissues of vertebrates. In fish, they appear in greater numbers in parts of their bodies that are exposed to their environment, such as skin, gills and intestine. The granules in fat snook's mast cell contain a variety of substances, such as histamine, heparin, chondroitin sulfate, serotonin, proteases and cytokines. The present study of intestine MCs/EGC was carried out in 20 specimens of fat snook. Samples of tissue were fixed in Bouin solution and in buffered formalin. Ferric hematoxylin - Congo red, pH6 acridine orange, pH2.5 and pH0,5 Alcian Blue (AB), toluidine blue, PAS, AB + PAS and immunohistochemistry protocols were used. In the mucosa and submucosa layers, MCs/EGCs granules with basic contents were evidenced by Congo red staining, and with acid contents granules were identified through pH 2.5 and 0,5 AB, and acridine orange. Basic and acid contents were simultaneously evidenced using ferric hematoxylin - Congo red stain. Metachromasia was observed in both mucosal and submucosal mast cells. Neutral glycoproteins were evidenced by using PAS protocol, glycosaminoglycan through AB and both simultaneously through AB + PAS. In immunohistochemistry assays, MCs/EGC were positive for tryptase, chymase and serotonin. As in mammals, the study of samples fixed in modified Karnovsky for transmission electron microscopy evidenced that most of the MCs granules were spherical and showed varying electron density, as described in previous reports on other teleost fish species. The metachromasia observed and the identification of tryptase, chymase and serotonin suggest a great similarity between fat snook's MCs/EGC and those described in the mucosa of mammals.

Does melatonin influence the apoptosis in rat uterus of animals exposed to continuous light?

Melatonin has been described as a protective agent against cell death and oxidative stress in different tissues, including in the reproductive system. However, the information on the action of this hormone in rat uterine apoptosis is low. Our objective was to evaluate the effects of melatonin on mechanisms of cell death in uterus of rats exposed to continuous light stress. Twenty adult Wistar rats were divided into two groups: GContr (vehicle control) and GExp which were treated with melatonin (0.4 mg/mL), both were exposed to continuous light for 90 days. The uterus was removed and processed for quantitative real time PCR (qRT-PCR), using PCR-array plates of the apoptosis pathway; for immunohistochemistry and TUNEL. The results of qRT-PCR of GEXP group showed up-regulation of 13 and 7, pro-apoptotic and anti-apoptotic genes, respectively, compared to GContr group. No difference in pro-apoptotic proteins (Bax, Fas and Faslg) expression was observed by immunohistochemistry, although the number of TUNEL-positive cells was lower in the group treated with melatonin compared to the group not treated with this hormone. Our data suggest that melatonin influences the mechanism and decreases the apoptosis in uterus of rats exposed to continuous light.

Differences in neonatal exposure to estradiol or testosterone on ovarian function and hormonal levels.

Exposure to an excess of androgen or estrogen can induce changes in reproductive function in adult animals that resemble polycystic ovary syndrome in humans. However, considerable differences exist among several types of animal models. Little is known about the molecular features of steroidogenesis and folliculogenesis in the ovaries of rats exposed to different sex steroids as neonates. Here, we evaluated the impact of androgen and estrogen exposure on the ovaries of adult female rats during their neonatal period in the gene expression of Lhr and Cyp17a1, two key players of steroidogenesis. We also assessed hormone levels, folliculogenesis and the theca-interstitial cell population. The study was performed on the second postnatal day in thirty female Wistar rats that were sorted into the following three intervention groups: testosterone, estradiol and vehicle (control group). The animals were euthanized 90 days after birth. The main outcomes were hormone serum levels, ovary histomorphometry and gene expression of Lhr and Cyp17a1 as analyzed via quantitative real-time PCR. We found that exposure to excess testosterone in early life increased the LH and testosterone serum levels, the LH/FSH ratio, ovarian theca-interstitial area and gene expression of Lhr and Cyp17a1 in adult rats. Estrogen induced an increase in the ovarian theca-interstitial area, the secondary follicle population and gene expression of Lhr and Cyp17a1. All animals exposed to the sex steroids presented with closed vaginas. Our data suggest that testosterone resulted in more pronounced reproductive changes than did estrogen exposure. Our results might provide some insight into the role of different hormones on reproductive development and on the heterogeneity of clinical manifestations of conditions such as polycystic ovary syndrome.

Lysosomal enzymes are decreased in the kidney of diabetic rats.

The objective of the present study was to investigate the expression and activities of lysosomal enzymes that act upon proteins and sulfated polysaccharides in diabetic rat kidney. Cathepsins, glycosidases and sulfatases were studied on the 10th (DM-10) and on the 30th (DM-30) day of streptozotocin-induced diabetes mellitus (DM). The activity of cathepsin B, the main kidney cysteine protease, was decreased both in DM-10 and DM-30. Gel filtration chromatography of urinary proteins has shown the prevalence of low molecular weight peptides in normal and DM-10 urine, in contrast to the prevalence of high molecular weight peptides and intact proteins in DM-30. These results show that the decrease in lysosomal proteases could explain, at least in part, the increased albuminuria detected by radial immunodiffusion (RID), due to the excretion of less degraded or intact albumin. Concerning sulfated polysaccharides, the activities of ß-glucuronidase, N-acetyl-ß-d-glucosaminidase, and N-acetyl-ß-d-galactosaminidase were also decreased in DM-30, while aryl sulfatases did not vary. Increased toluidine blue metachromatic staining of the tissue suggests that the lower activities of glycosidases could lead to intracellular deposition of partially digested molecules, and this could explain the decreased urinary excretion and increased tissue buildup of these molecules. The main morphological changes observed in kidney were proximal convoluted tubules with thinner walls and thinner brush border. Immunohistochemistry revealed that most of cathepsin B was located in the brush border of proximal tubular cells, highlighting the involvement of proximal convoluted tubules in diabetic nephropathy.

Hormonal and Metabolic Factors Influence the Action of Progesterone on the Endometrium of Women with Polycystic Ovary Syndrome.

Hormonal and metabolic factors may influence endometrial quality and interfere with the action of progesterone. Therefore, the aim of our study was to address this issue. Participants were recruited from an outpatient reproductive endocrinology clinic at an academic tertiary medical care centre. All subjects underwent endometrial biopsy (EB) in the follicular phase of the cycle prior to treatment. Thereafter, they were treated with micronized progesterone (400 mg/day × 10 days intravaginally) from days 14-28 of the next cycle. A second EB was performed between days 21-24 of the cycle (the second phase). The metabolic and hormonal serum levels were evaluated during the implantation window. EB samples were analysed using light microscopy for histomorphometric analysis. The endometrium of women with Polycystic Ovarian Syndrome (PCOS) in the second phase demonstrated a uniform surface epithelium with less leukocyte infiltration and an absence of apoptotic figures compared to the control group. (p < 0.021). The thickness of the surface epithelium in the second phase of the PCOS group correlated positively with free and bioavailable testosterone values. The number of stromal cells increases with increasing insulin levels. Our results suggest that histomorphometric abnormalities of the endometrium persist and are linked to androgen and insulin levels despite progesterone supplementation in PCOS.

Influence of estrogen replacement and aging on the expression of nerve growth factor in the urethra of female rats.

OBJECTIVE: To evaluate the expression of nerve growth factor (NGF) in the urethra of adult female rats in different hormonal status using immunohistochemical assay. METHODS: Forty-eight rats (Rattus norvegicus albinus, Rodentia, Mammalia) from the CEDEME-UNIFESP laboratory animal facility were used in the study. Rats were divided into four groups: group A, 12 non-neutered rats; group B, 12 oophorectomized rats; group C, 12 castrated rats treated with 17ß-estradiol for 30 days; and group D, 12 aging rats. Animals were killed by lethal injection and their urethra was removed. NGF expression was evaluated by means of immunohistochemistry using mouse monoclonal primary IgG antibody anti-NGF diluted 1:600, and read under 400× magnification. Digital analysis of the images was done by Imagelab software. The intensity of the dark brown color was used as a measure of NGF cytoplasmatic expression, and was used to quantify the percentage of epithelial and muscular layer cells showing this neurotrophin. RESULTS: After oophorectomy, rats showed a significant increase in NGF expression in the periurethral muscular layer. Compared with oophorectomized rats, NGF expression increased in the epithelial layer and diminished in the periurethral smooth muscle following estrogen administration. In 18-month-old rats, NGF expression was diminished in both epithelial and muscular layers. CONCLUSIONS: Hormonal status led to significant differences in NGF protein expression in urethral epithelium and periurethral smooth muscle.

Effects of lidocaine with epinephrine or with buffer on wound healing in rat skin.

Lidocaine blocks nociceptive fibers, preventing initial wound signaling and mast cell degranulation. It is hypothesized that epinephrine and buffer affect the wound healing by potentiating lidocaine blockage. This double-blind, randomized, self-controlled study investigated this possibility using male Wistar rats, which were injected with 2 mL of different solutions into the left and right sides of the back. Then, each side was incised and sutured. Sixty rats were divided in three groups: saline solution (SS) and lidocaine; lidocaine and lidocaine with buffer; lidocaine with epinephrine and lidocaine with epinephrine and buffer. Half of each group was sacrificed 7 days after surgery and the remaining after 21 days. A sample of each wound was obtained and quantified for the level of collagen present using computer morphometry and for mast cell quantity. There were no differences between animals with regard to the collagen. However, mast cell levels in the same animal significantly differed between SS × lidocaine. Comparison of the same injected substance between animals with different healing dates showed a significant effect on collagen SS and on all mast cells, except SS. Lidocaine affected collagenization and decreased the initial quantity of mast cells at the wound site.

Effects of hexestrol on mouse ovarian morphology and ovulation.

OBJECTIVE: To analyze histological aspects of ovaries as well as the ovulation of adult mice treated with the anabolic agent hexestrol. METHODS: Thirty adult mice were divided into three groups of 10 animals each: (GI) the animals received a dose of 3 mg/kg of hexestrol; (GII) the animals were given a dose of 6 mg/kg of hexestrol; (GIII) the animals were injected with distilled water (vehicle). Another 10-animal group (GIV) was included, and these mice were injected with propionate testosterone (1.25 mg) after 5 days from the day of birth. Hexestrol was administered intraperitoneally once a day and the treatment lasted 30 days. The mice were then sacrificed; their ovaries and oviducts were removed, submitted to histological routine and analyzed under light microscopy. RESULTS: In mice treated with hexestrol (6 mg/kg) (Group II), ovaries were smaller than those from the controls but highly vascularized; similar results were obtained in GIV. A great number of follicles in several stages of development were found -- however, with no corpora lutea -- in six animals in GII. No corpora lutea were found in GIV. The number of luteal bodies and oocytes in GII was lower than that in GI or GIII. No oocytes were detected in GIV. Finally, the nuclear volume of interstitial cells in GII and GIV was the largest. CONCLUSION: Our data suggest that the anabolic agent hexestrol in a high dose may decrease ovulation in mice.

Effects of estrogen status in osteocyte autophagy and its relation to osteocyte viability in alveolar process of ovariectomized rats.

Estrogen maintains osteocyte viability, whereas its deficiency induces osteocyte apoptosis. As autophagy is important for osteocyte viability, we hypothesized whether the anti-apoptotic effect of estrogen is related to autophagy in osteocytes. Thirty adult female rats were sham-operated (SHAM) or ovariectomized (OVX). After three weeks, twelve rats of SHAM and OVX groups were killed before treatment (basal period), whereas the remaining rats received estrogen (OVXE) or vehicle (OVX) for 45 days. Fragments of maxilla containing alveolar process of the first molars were embedded in paraffin or Araldite. Paraffin-sections were stained with hematoxylin/eosin for histomorphometry, or subjected to the silver impregnation method for morphological analysis of osteocyte cytoplasmic processes. Autophagy was analyzed by immunohistochemical detections of beclin-1, MAP-LC3α and p62, whereas apoptosis was evaluated by immunohistochemical detections of cleaved caspase-3 and BAX, TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) method and by ultrastructural analysis. Araldite-semithin sections were subjected to the Sudan-black method for detection of lipids. OVX-basal group showed high frequency of caspase-3-, TUNEL- and p62-positive osteocytes accompanied with low frequency of beclin-1- and MAP-LC3α-positive osteocytes. At 45 days, OVXE group exhibited higher number of osteocytes, higher frequency of beclin-1- and MAP-LC3α-positive osteocytes, and lower frequency of caspase-3, BAX-, TUNEL- and p62-positive osteocytes than OVX group. Significant reduction in bone area was observed in the OVX compared to OVXE and SHAM groups. The highest frequency of Sudan-Black-positive osteocytes and osteocytes with scarce cytoplasmic processes, or showing apoptotic features were mainly observed in OVX groups. Our results indicate that estrogen deficiency decreases autophagy and increases apoptosis, whereas estrogen replacement enhances osteocyte viability by inhibiting apoptosis and maintaining autophagy in alveolar process osteocytes. These results suggest that the anti-apoptotic effect of estrogen may be, at least in part, related to autophagy regulation in osteocytes.

Effects of topical estradiol on the facial skin collagen of postmenopausal women under oral hormone therapy: a pilot study.

OBJECTIVE: To analyze the dermal collagen of 15 postmenopausal women who had being treated with systemic estrogen replacement before and after using topical a 0.01% estrogen treatment. METHODS: Fifteen patients were included in this clinical trial using the systemic estrogen therapy for at least 1 year (minimum and maximum lengths of therapy were 13 and 40 months, respectively). A facial punch was performed in the preauricular area for collecting samples before and after the 16 weeks of treatment. Blood samples were also collected for estradiol level determination. The morphometric determination of epithelial and dermal thickness as well as dermal collagen were measured using a suitable software. The paired Student's t-test was used for statistical analysis. RESULTS: The epithelial and dermal thickness enhanced after the topic estrogen therapy (P<0.01). The amount of collagen significantly increased after 16 weeks of treatment (P<0.001). The estrogen levels did not significant increase after the topical therapy (P > or = 0.05). CONCLUSION: Our data suggested that topical estrogen associated to systemic estrogen therapy seems to increase the expression of skin collagen amount, which may prove to be beneficial for the postmenopausal facial skin.

Small leucine-rich proteoglycans (SLRPs) in the endometrium of polycystic ovary syndrome women: a pilot study.

BACKGROUND: Small leucine-rich proteoglycans (SLRPs) play an important role in tissue homeostasis and cell proliferation since these proteoglycans sequester multiple growth factors. However, the content of SLRPs in the endometrium of polycystic ovary syndrome (PCOS) women is unknown. Our purpose was to test the hypothesis that excessive endometrial proliferation in PCOS may be partly related to abnormalities in SLRPs. METHODS: In a cross section study a total of 20 endometrial samples were collected from 10 patients with PCOS and 10 ovulatory women during their proliferative (pre-ovulatory) phase. The study subjects were matched for age, body mass index and race. The age range was 20 to 35 years. All volunteers were evaluated in reproductive endocrinology clinic, Gynecology Division, Clinics Hospital, University of São Paulo Medical School Profile and concentration of small leucine-rich proteoglycans (decorin, lumican, fibromodulin and biglycan) were determined by immunohistochemical testing and Western blotting. RESULTS: Decorin and lumican demonstrated higher immunoreactivity and relative expression in the endometrium of women with PCOS compared to that of women with regular menstrual cycles. CONCLUSION: Our data suggests that the endometrium of PCOS women demonstrate a greater content of SLRP than controls; decorin and lumican, in particular, were found in higher concentrations in the endometrium of PCOS women during the proliferative phase. These differences may, in part, explain the excess of endometrial proliferation frequently observed in PCOS. Further studies are warranted.

Impaired branched-chain amino acid metabolism may underlie the nonalcoholic fatty liver disease-like pathology of neonatal testosterone-treated female rats.

Polycystic ovary syndrome (PCOS) is frequently associated with non-alcoholic fatty liver disease (NAFLD), but the mechanisms involved in the development of NAFLD in PCOS are not well known. We investigated histological changes and metabolomic profile in the liver of rat models of PCOS phenotype induced by testosterone or estradiol. Two-day old female rats received sc injections of 1.25 mg testosterone propionate (Testos; n = 10), 0.5 mg estradiol benzoate (E2; n = 10), or vehicle (control group, CNT; n = 10). Animals were euthanized at 90-94 d of age and the liver was harvested for histological and metabolomic analyses. Findings showed only Testos group exhibited fatty liver morphology and higher levels of ketogenic and branched-chain amino acids (BCAA). Enrichment analysis showed effects of testosterone on BCAA degradation pathway and mitochondrial enzymes related to BCAA metabolism. Testos group also had a decreased liver fatty acid elongase 2 (ELOVL2) activity. E2 group had reduced lipid and acylcarnitine metabolites in the liver. Both groups had increased organic cation transporters (SLC22A4 and SLC16A9) activity. These findings indicate that neonatal testosterone treatment, but not estradiol, produces histological changes in female rat liver that mimic NAFLD with testosterone-treated rats showing impaired BCAA metabolism and dysfunctions in ELOVL2, SLC22A4 and SLC16A9 activity.

Morphofunctional alterations of the nonpregnant murine uterus in response to intense and exhaustive exercise are not related to oxidative stress.

Exercise is a common and noninvasive way to improve human health. In contrast, intense exercise causes damage in various tissues and is usually associated with metabolic changes in organs and tissues. Even though intense exercise is associated with dysfunctions in the female reproductive system, much less is known about the cellular mechanisms underlying its effects particularly on the nonpregnant uterus. We investigated whether the effects of an intense and exhaustive exercise (IEE) program on the isolated C57BL/6 uterine morphology and contractility might be related to increased levels of prooxidation markers. Female mice were submitted to 2 days of IEE. The daily exercise session consisted of a running session until exhaustion, with the treadmill speed set at 85% of each animal's maximum velocity. Training responses were evaluated through two parameters: time to exhaustion and maximum velocity. Absence of exercise-induced hypothalamic-pituitary-adrenal (HPA) axis activation was indirectly evaluated by maintenance of the adrenal gland weight. IEE reduced the thickness of the longitudinal muscular layer by 10%, impaired contractility in response to muscarinic stimulation (increased EC50 and decreased Emax), but showed a strong trend to decreasing the KCl-induced contraction; reduced lipid peroxidation; and did not alter the uterine protein oxidation of exercised animals compared with control. Altogether we provide evidence for the nonpregnant murine uterus being an important target to IEE, leading to morphofunctional alterations which could not be associated with tissue oxidative stress but might well be related with exercise-induced uterine dysfunctions.

Melatonin influences on steroidogenic gene expression in the ovary of pinealectomized rats.

OBJECTIVE: To analyze the expression of genes related to steroidogenesis in the ovary of pinealectomized rats. DESIGN: Experimental research. SETTING: University research laboratory. ANIMAL(S): Thirty female adult rats. INTERVENTION(S): Administration of vehicle (GI), pinealectomy with vehicle (GII), or pinealectomy with melatonin replacement (10 µg/night) for 60 consecutive days (GIII), then euthanasia after 2 months of treatment, ovary collection complementary DNA microarray analyses, confirmatory quantitative reverse-transcriptase polymerase chain reaction analyses, and immunohistochemical analyses for localizing steroidogenesis changes in the ovary. MAIN OUTCOME MEASURE(S): Biologic molecular study followed by immunohistochemical analysis. RESULT(S): The changes in the expression of CYP11A1, CYP17A1, and CYP19A1 after pinealectomy (GII) compared with control (GI) showed the Cyp17a1 expression level increased in the theca interna and interstitial cells in the GII rats compared with the other groups. CONCLUSION(S): Melatonin deprivation (pinealectomy) or administration may influence the ovarian CYP17A1 expression and steroidogenesis.

Heme oxygenase 1 improves glucoses metabolism and kidney histological alterations in diabetic rats.

One important concern in the treatment of diabetes is the maintenance of glycemic levels and the prevention of diabetic nephropathy. Inducible heme oxygenase 1 (HO-1) is a rate-limiting enzyme thought to have antioxidant and cytoprotective roles. The goal of the present study was to analyze the effect of HO-1 induction in chronically hyperglycemic rats. The hyperglycemic rats were divided into two groups: one group, called STZ, was given a single injection of streptozotocin; and the other group was given a single streptozotocin injection as well as daily injections of hemin, an HO-1 inducer, over 60 days (STZ + HEME). A group of normoglycemic, untreated rats was used as the control (CTL).Body weight, diuresis, serum glucose levels, microalbuminuria, creatinine clearance rate, urea levels, sodium excretion, and lipid peroxidation were analyzed. Histological alterations and immunohistochemistry for HO-1 and inducible nitric oxide synthase (iNOS) were assessed. After 60 days, the STZ group exhibited an increase in blood glucose, diuresis, urea, microalbuminuria, and sodium excretion. There was no weight gain, and there was a decrease in creatinine clearance in comparison to the CTL group. In the STZ + HEME group there was an improvement in the metabolic parameters and kidney function, a decrease in blood glucose, serum urea, and microalbuminuria, and an increase of creatinine clearance, in comparison to the STZ group.There was glomerulosclerosis, collagen deposition in the STZ rats and increase in iNOS and HO-1 expression. In the STZ + HEME group, the glomerulosclerosis and fibrosis was prevented and there was an increase in the expression of HO-1, but decrease in iNOS expression and lipid peroxidation. In conclusion, our data suggest that chronic induction of HO-1 reduces hyperglycemia, improves glucose metabolism and, at least in part, protects the renal tissue from hyperglycemic injury, possibly through the antioxidant activity of HO-1.