Long-term Melatonin Treatment Reduces Ovarian Mass And Enhances Tissue Antioxidant Defenses During Ovulation In The Rat.

Melatonin regulates the reproductive cycle, energy metabolism and may also act as a potential antioxidant indoleamine. The present study was undertaken to investigate whether long-term melatonin treatment can induce reproductive alterations and if it can protect ovarian tissue against lipid peroxidation during ovulation. Twenty-four adult female Wistar rats, 60 days old (± 250-260 g), were randomly divided into two equal groups. The control group received 0.3 mL 0.9% NaCl + 0.04 mL 95% ethanol as vehicle, and the melatonin-treated group received vehicle + melatonin (100 µg·100 g body weight(-1)·day(-1)) both intraperitoneally daily for 60 days. All animals were killed by decapitation during the morning estrus at 4:00 am. Body weight gain and body mass index were reduced by melatonin after 10 days of treatment (P < 0.05). Also, a marked loss of appetite was observed with a fall in food intake, energy intake (melatonin 51.41 ± 1.28 vs control 57.35 ± 1.34 kcal/day) and glucose levels (melatonin 80.3 ± 4.49 vs control 103.5 ± 5.47 mg/dL) towards the end of treatment. Melatonin itself and changes in energy balance promoted reductions in ovarian mass (20.2%) and estrous cycle remained extensive (26.7%), arresting at diestrus. Regarding the oxidative profile, lipid hydroperoxide levels decreased after melatonin treatment (6.9%) and total antioxidant substances were enhanced within the ovaries (23.9%). Additionally, melatonin increased superoxide dismutase (21.3%), catalase (23.6%) and glutathione-reductase (14.8%) activities and the reducing power (10.2% GSH/GSSG ratio). We suggest that melatonin alters ovarian mass and estrous cyclicity and protects the ovaries by increasing superoxide dismutase, catalase and glutathione-reductase activities.44217-2

Ovarian histology and follicular score in female rats treated with nandrolone decanoate and submitted to physical effort

The study was conducted to analyze the histology of the ovaries of adults rats treated with steroids, and submitted or not to physical effort. The control group consisted of females submitted to physical effort and sedentary females, both of which received a physiological solution of 0.9% saline. Treated females, sedentary or not, received 6 mg/kg of body weight of nandrolone decanoate. The steroid and physiological solution were administered intraperitoneally, with a single injection per week for 4 consecutive weeks. The applied physical effort was swimming (20 minutes daily, 5 days/week, for the 4 weeks of treatment). Serial sections (5 μm) of ovaries were prepared for histological evaluation and follicular score. The weight of ovaries and hypophysis, the number of antral and atretic follicles, and the area of corpus luteum were all affected by the steroids. In the ovaries of the control groups, well-developed corpus luteum was observed. In the treated groups, the cortical stroma was occupied by ovarian interstitial tissue. The females treated with steroids presented estral acyclicity. The use of nandrolone decanoate, whether associated with physical effort or not, affected the morphological pattern of the ovaries

OVARIAN HISTOLOGY and FOLLICULAR SCORE IN FEMALE RATS TREATED WITH NANDROLONE DECANOATE and SUBMITTED TO PHYSICAL EFFORT

The study was conducted to analyze the histology of the ovaries of adults rats treated with steroids, and submitted or not to physical effort. The control group consisted of females submitted to physical effort and sedentary females, both of which received a physiological solution of 0.9% saline. Treated females, sedentary or not, received 6 mg/kg of body weight of nandrolone decanoate. The steroid and physiological solution were administered intraperitoneally, with a single injection per week for 4 consecutive weeks. The applied physical effort was swimming (20 minutes daily, 5 days/week, for the 4 weeks of treatment). Serial sections (5 mu m) of ovaries were prepared for histological evaluation and follicular score. The weight of ovaries and hypophysis, the number of antral and atretic follicles, and the area of corpus luteum were all affected by the steroids. In the ovaries of the control groups, well-developed corpus luteum was observed. In the treated groups, the cortical stroma was occupied by ovarian interstitial tissue. The females treated with steroids presented estral acyclicity. The use of nandrolone decanoate, whether associated with physical effort or not, affected the morphological pattern of the ovaries

Serum miRNAs are differentially altered by ethanol and caffeine consumption in rats

Alcoholism is a multifactorial disease with high risk for dependence determined by genetic background, environmental factors and neuroadaptations. The excessive consumption of this substance is related to psychiatric problems, epilepsy, cardiovascular disease, cirrhosis and cancers. Caffeine is one of the most popular psychostimulants currently consumed in the world. The combination of ethanol and caffeine ingested by consuming “energy drinks” is becoming increasingly popular among young people. We analyzed the effect of simultaneous consumption of ethanol and caffeine on the serum profile of miRNAs differentially expressed in the ethanol-drinking rat model (UChB strain). Adult rats were divided into three groups (n = 5 per group): UChB group (rats fed with 1 : 10 (v/v) ethanol ad libitum); UChB + caffeine group (rats fed with 1 : 10 (v/v) ethanol ad libitum + 3 g L−1 of caffeine); control group (rats drinking water used as the control for UChB). The treatment with caffeine occurred from day 95 to 150 days old, totalizing 55 days of ethanol + caffeine ingestion. The expressions of microRNAs (miR) -9-3p, -15b-5p, -16-5p, -21-5p, -200a-3p and -222-3p were detected by Real Time-PCR (RT-PCR). The expressions of miR-9-3p, -15b-5p, -16-5p and -222-3p were upregulated in the UChB group. Conversely, simultaneous ingestion of ethanol and caffeine significantly reversed these expressions to similar levels to control animals, thus emphasizing that caffeine had a protective effect in the presence of ethanol. In addition, miR-21-5p was downregulated with ethanol consumption whereas miR-222-3p was unchanged. Ethanol and caffeine consumption was capable of altering serum miRNAs, which are potential biomarkers for the systemic effects of these addictive substances86842849FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2013/11095-2; 2014/06240-

Determination of stanozolol and 3[prime]-hydroxystanozolol in rat hair, urine and serum using liquid chromatography tandem mass spectrometry

The developed methods are sensitive, specific and reproducible for the determination of stanozolol and 3'-hydroxystanozolol in rat hair, urine and serum. These methods can be used for in vivo studies further investigating stanozolol metabolism, but also could be extended for doping testing. Owing to the complementary nature of these tests, with urine and serum giving information on recent drug use and hair providing retrospective information on habitual use, it is suggested that blood or urine tests could accompany hair analysis and thus avoid false doping results

Melatonin Attenuates The Tlr4-mediated Inflammatory Response Through Myd88- And Trif-dependent Signaling Pathways In An In Vivo Model Of Ovarian Cancer

Background: Toll-like receptors (TLRs) are effector molecules expressed on the surface of ovarian cancer (OC) cells, but the functions of the TLR2/TLR4 signaling pathways in these cells remain unclear. Melatonin (mel) acts as an anti-inflammatory factor and has been reported to modulate TLRs in some aggressive tumor cell types. Therefore, we investigated OC and the effect of long-term mel therapy on the signaling pathways mediated by TLR2 and TLR4 via myeloid differentiation factor 88 (MyD88) and toll-like receptor-associated activator of interferon (TRIF) in an ethanol-preferring rat model. Methods: To induce OC, the left ovary of animals either consuming 10% (v/v) ethanol or not was injected directly under the bursa with a single dose of 100μg of 7,12-dimethylbenz(a)anthracene (DMBA) dissolved in 10 μL of sesame oil. The right ovaries were used as sham-surgery controls. After developing OC, half of the animals received i.p. injections of mel (200 μg/100 g b.w./day) for 60 days. Results: Although mel therapy was unable to reduce TLR2 levels, it was able to suppress the OC-associated increase in the levels of the following proteins: TLR4, MyD88, nuclear factor kappa B (NFkB p65), inhibitor of NFkB alpha (IkBα), IkB kinase alpha (IKK-α), TNF receptor-associated factor 6 (TRAF6), TRIF, interferon regulatory factor 3 (IRF3), interferon β (IFN-β), tumor necrosis factor alpha (TNF-α), and interleukin (IL)-6. In addition, mel significantly attenuated the expression of IkBα, NFkB p65, TRIF and IRF-3, which are involved in TLR4-mediated signaling in OC during ethanol intake. Conclusion: Collectively, our results suggest that mel attenuates the TLR4-induced MyD88- and TRIF-dependent signaling pathways in ethanol-preferring rats with OC.151Cannistra, S.A., Cancer of the ovary (2004) N Engl J Med, 351, pp. 2519-2565Fallows, S., Price, J., Atkinson, R.J., Johnston, P.G., Hickey, I., Russell, S.E., P53 mutation does not affect prognosis in ovarian epithelial malignancies (2001) J Pathol, 194, pp. 68-75Kelly, M.G., Alvero, A.B., Chen, R., Silasi, D.A., Abrahams, V.M., Chan, S., TLR-4 signaling promotes tumor growth and paclitaxel chemoresistance in ovarian cancer (2006) Cancer Res, 66, pp. 3859-3868Chen, R., Alvero, A.B., Silasi, D.A., Steffensen, K.D., Mor, G., Cancers take their toll-the function and regulation of toll-like receptors in cancer cells (2008) Oncogene, 27, pp. 225-233Szajnik, M., Szczepanski, M.J., Czystowska, M., Elishaev, E., Mandapathil, M., Nowak-Markwitz, E., TLR4 signaling induced by lipopolysaccharide or paclitaxel regulates tumor survival and chemoresistance in ovarian cancer" (2009) Oncogene, 28, pp. 4353-4363Wang, A.C., Su, Q.B., Wu, F.X., Zhang, X.L., Liu, P.S., Role of TLR4 for paclitaxel chemotherapy in human epithelial ovarian cancer cells (2009) Eur J Clin Invest, 39, pp. 157-164Hsu, D., Fukata, M., Hernandez, Y.G., Sotolongo, J.P., Goo, T., Maki, J., Toll-like receptor 4 differentially regulates epidermal growth factor-related growth factors in response to intestinal mucosal injury (2010) Lab Invest, 90, pp. 1295-1305Beinke, S., Ley, S.C., Functions of NF-kappaB1 and NF-kappaB2 in immune cell biology (2004) Biochem J, 382, pp. 393-409Akira, S., Toll-like receptor signaling (2003) J Biol Chem, 278, pp. 38105-38108Akira, S., Takeda, K., Toll-like receptor signalling (2004) Nat Rev Immunol, 4, pp. 499-511French, S.W., Oliva, J., French, B.A., Li, J., Bardag-Gorce, F., Alcohol, nutrition and liver cancer: role of Toll-like receptor signaling (2010) World J Gastroenterol, 16, pp. 1344-1348Chuffa, L.G., Fioruci-Fontanelli, B.A., Mendes, L.O., Fávaro, W.J., Pinheiro, P.F., Martinez, M., Characterization of chemically induced ovarian carcinomas in an ethanol-preferring rat model: influence of long-term melatonin treatment (2013) PLoS One, 8, p. e81676Ferreira, G.M., Martinez, M., Camargo, I.C., Domeniconi, R.F., Martinez, F.E., Chuffa, L.G., Melatonin attenuates Her-2, p38 MAPK, p-AKT, and mTOR levels in ovarian carcinoma of ethanol-preferring rats (2014) J Cancer Educ, 5, pp. 728-735Stehle, J.H., Saade, A., Rawashdeh, O., Ackermann, K., Jilg, A., Sebestény, T., A survey of molecular details in the human pineal gland in the light of phylogeny, structure, function and chronobiological diseases (2011) J Pineal Res, 51, pp. 17-43Blask, D.E., Dauchy, R.T., Sauer, L.A., Putting cancer to sleep at night: the neuroendocrine/circadian melatonin signal (2005) Endocrine, 27, pp. 179-188Cos, S., González, A., Martínez-Campa, C., Mediavilla, M.D., Alonso-González, C., Sánchez-Barceló, E.J., Melatonin as a selective estrogen enzyme modulator (2008) Curr Cancer Drug Targets, 8, pp. 691-702Petranka, J., Baldwin, W., Biermann, J., Jayadev, S., Barrett, J.C., Murphy, E., The oncostatic action of melatonin in an ovarian carcinoma cell line (1999) J Pineal Res, 26, pp. 129-136Reiter, R.J., Mechanisms of cancer inhibition by melatonin (2004) J Pineal Res, 3, pp. 213-214Calvo, J.R., González-Yanes, C., Maldonado, M.D., The role of melatonin in the cells of the innate immunity: a review (2013) J Pineal Res, 55, pp. 103-120Hu, Z.P., Fang, X.L., Fang, N., Wang, X.B., Qian, H.Y., Cao, Z., Melatonin ameliorates vascular endothelial dysfunction, inflammation, and atherosclerosis by suppressing the TLR4/NF-jB system in high-fat-fed rabbits (2013) J Pineal Res, 55, pp. 388-398Tamura, E.K., Cecon, E., Monteiro, A.W., Silva, C.L., Markus, R.P., Melatonin inhibits LPS-induced NO production in rat endothelial cells (2009) J Pineal Res, 46, pp. 268-274Mauriz, J.L., Collado, P.S., Veneroso, C., Reiter, R.J., González-Gallego, J., A review of the molecular aspects of melatonin's anti-inflammatory actions: recent insights and new perspectives (2013) J Pineal Res, 54, pp. 1-14Conti, A., Maestroni, G.J., The clinical neuroimmunotherapeutic role of melatonin in oncology (1995) J Pineal Res, 19, pp. 103-110Chada, S., Ramesh, R., Mhashilkar, A.M., Cytokine- and chemokine-based gene therapy for cancer (2003) Curr Opin Mol Ther, 5, pp. 463-474Kang, J.W., Koh, E.J., Lee, S.M., Melatonin protects liver against ischemia and reperfusion injury through inhibition of toll-like receptor signaling pathway (2011) J Pineal Res, 50, pp. 403-411Chuffa, L.G., Amorim, J.P., Teixeira, G.R., Mendes, L.O., Fioruci, B.A., Pinheiro, P.F., Long-term exogenous melatonin treatment modulates overall feed efficiency and protects ovarian tissue against injuries caused by ethanol-induced oxidative stress in adult UChB rats (2011) Alcohol Clin Exp Res, 35, pp. 1498-1508Chuffa, L.G., Seiva, F.R., Fávaro, W.J., Amorim, J.P., Teixeira, G.R., Mendes, L.O., Melatonin and ethanol intake exert opposite effects on circulating estradiol and progesterone and differentially regulate sex steroid receptors in the ovaries, oviducts, and uteri of adult rats (2013) Reprod Toxicol, 39, pp. 40-49Chuffa, L.G., Seiva, F.R., Fávaro, W.J., Teixeira, G.R., Amorim, J.P., Mendes, L.O., Melatonin reduces LH, 17 beta-estradiol and induces differential regulation of sex steroid receptors in reproductive tissues during rat ovulation (2011) Reprod Biol Endocrinol, 9, p. 108Chuffa, L.G., Amorim, J.P., Teixeira, G.R., Mendes, L.O., Fioruci, B.A., Pinheiro, P.F., Long-term melatonin treatment reduces ovarian mass and enhances tissue antioxidant defenses during ovulation in the rat (2011) Braz J Med Biol Res, 44, pp. 217-223Hoyer, P.B., Davis, J.R., Bedrnicek, J.B., Marion, S.L., Christian, P.J., Barton, J.K., Ovarian neoplasm development by 7,12-dimethylbenz[a]anthracene (DMBA) in a chemically-induced rat model of ovarian failure (2009) Gynecol Oncol, 112, pp. 610-615Zhou, M., McFarland-Mancini, M.M., Funk, H.M., Husseinzadeh, N., Mounajjed, T., Drew, A.F., Toll-like receptor expression in normal ovary and ovarian tumors (2009) Cancer Immunol Immunother, 58, pp. 1375-1385Xia, M.Z., Liang, Y.L., Wang, H., Chen, X., Huang, Y.Y., Zhang, Z.H., Melatonin modulates TLR4-mediated inflammatory genes through MyD88- and TRIF-dependent signaling pathways in lipopolysaccharide-stimulated RAW264.7 cells (2012) J Pineal Res, 53, pp. 325-334Lucas, K., Maes, M., Role of the Toll Like receptor (TLR) radical cycle in chronic inflammation: possible treatments targeting the TLR4 pathway (2013) Mol Neurobiol, 48, pp. 190-204Fernandez-Lizarbe, S., Montesinos, J., Guerri, C., Ethanol induces TLR4/TLR2 association, triggering an inflammatory response in microglial cells (2013) J Neurochem, 126, pp. 261-273O'Neill, L.A., Fitzgerald, K.A., Bowie, A.G., The Toll-IL-1 receptor adaptor family grows to five members (2003) Trends Immunol, 24, pp. 286-290Wang, Z., Wu, L., You, W., Ji, C., Chen, G., Melatonin alleviates secondary brain damage and neurobehavioral dysfunction after experimental subarachnoid hemorrhage: possible involvement of TLR4-mediated inflammatory pathway (2013) J Pineal Res, 55, pp. 399-408Wagnerberger, S., Fiederlein, L., Kanuri, G., Stahl, C., Millonig, G., Mueller, S., Sex-specific differences in the development of acute alcohol-induced liver steatosis in mice (2013) Alcohol Alcohols, 48, pp. 648-656Natoli, G., Chiocca, S., Nuclear ubiquitin ligases, NF-kappaB degradation, and the control of inflammation. (2008) Sci Signal, 1Barlin, J.N., Jelinic, P., Olvera, N., Bogomolniy, F., Bisogna, M., Dao, F., Validated gene targets associated with curatively treated advanced serous ovarian carcinoma (2013) Gynecol Oncol, 128, pp. 512-517Landen, C.N., Birrer, M.J., Sood, A.K., Early events in the pathogenesis of epithelial ovarian cancer (2008) J Clin Oncol, 26, pp. 995-1005Kim, J.M., Kim, S.H., Ko, S.H., Jung, J., Chun, J., Kim, N., The guggulsterone derivative GG-52 inhibits NF-ΚB signaling in gastric epithelial cells and ameliorates ethanol-induced gastric mucosal lesions in mice (2013) Am J Physiol Gastrointest Liver Physiol, 304, pp. G193-202Blanco, A.M., Guerri, C., Ethanol intake enhances inflammatory mediators in brain: role of glial cells and TLR4/IL-1RI receptors (2007) Front Biosci, 12, pp. 2616-2630Kim, W.H., Hong, F., Jaruga, B., Hu, Z., Fan, S., Liang, T.J., Additive activation of hepatic NF-Κ B by ethanol and HBX or HCV core protein: involvement of TNF-α receptor I-independent and -dependent mechanisms (2001) FASEB J, 15, pp. 2551-2553Sato, S., Sugiyama, M., Yamamoto, M., Watanabe, Y., Kawai, T., Takeda, K., Toll/IL-1 receptor domain-containing adaptor inducing IFN-beta (TRIF) associates with TNF receptor-associated factor 6 and TANK-binding kinase 1, and activates two distinct transcription factors, NF-kappa B and IFN-regulatory factor-3, in the Toll-like receptor signaling (2003) J Immunol, 171, pp. 4304-4310Fitzgerald, K.A., McWhirter, S.M., Faia, K.L., Rowe, D.C., Latz, E., Golenbock, D.T., IKKepsilon and TBK1 are essential components of the IRF3 signaling pathway (2003) Nat Immunol, 4, pp. 491-496Muccioli, M., Sprague, L., Nandigam, H., Pate, M., Benencia, F., Toll-like receptors as novel therapeutic targets for ovarian cancer (2012) ISRN Oncol, 2012, p. 642141Petrasek, J., Dolganiuc, A., Csak, T., Nath, B., Hritz, I., Kodys, K., Interferon regulatory factor 3 and type I interferons are protective in alcoholic liver injury in mice by way of crosstalk of parenchymal and myeloid cells (2011) Hepatology, 53, pp. 649-660Ruegg, C., Leukocytes, inflammation, and angiogenesis in cancer: fatal attractions (2006) J Leukoc Biol, 80, pp. 682-684Santin, A.D., Hermonat, P.L., Ravaggi, A., Cannon, M.J., Pecorelli, S., Parham, G.P., Secretion of vascular endothelial growth factor in ovarian cancer (1999) Eur J Gynaecol Oncol, 20, pp. 177-181Garcia-Mauriño, S., Gonzalez-Haba, M.G., Calvo, J.R., Rafii-El-Idrissi, M., Sanchez-Margalet, V., Goberna, R., Melatonin enhances IL-2, IL-6, and IFN-gamma production by human circulating CD4+ cells: A possible nuclear receptor-mediated mechanism involving Thelper type 1 lymphocytes and monocytes (1997) J Immunol, 159, pp. 574-58