Ginger Essential Oil Ameliorates Cisplatin-Induced Nephrotoxicity in Mice

Purpose: To investigate the effect of ginger essential oil (GEO) in an experimental model of cisplatininduced nephrotoxicity.Methods: Male mice were divided into treatment six groups (n = 7), namely: Groups I (saline), II and III (cisplatin, 10 mg/kg, i.p.) euthanized in 3th and 6th days, respectively, and IV, V and IV (GEO, 100, 200 and 400 mg/kg/day, respectively, by gavage 3, 4, 5 and 6 days after cisplatin injection). Creatinine levels and protein/creatinine ratio were determined in plasma and urine, respectively. Bone morphogenic protein (BMP-7) and tumor necrosis factor (TNF-α) levels of kidney tissues were determined while mRNA expression levels were obtained using real-time polymerase chain reaction.Results: GEO treatment reduced significantly creatinine levels to 0.53 ± 0.02; 0.48 ± 0.008 and 0.46 ± 0.02 at 100, 200 and 400 mg/kg, respectively, compared with control (0.70 ± 0.01) [p<0.05] but increased protein : creatinine ratio to 0.21 ± 0.01, 0.22 ± 0.01, 0.24 ± 0.02 compared with control (0.06 ± 0.008) [p<0.05]. Pro-inflammatory TNF-α mRNA expression was decreased to 1.46 ± 0.21, 1.39 ± 0.19 and 1.36 ± 0.09, at GEO doses of 100, 200 and 400 mg/kg, respectively, while anti-fibrotic BMP-7 mRNA expression increased to 2.05 ± 0.26 and 2.44 ± 0.42 at doses of 200 and 400 mg/kg, respectively, compared with control (0.59 ± 0.39, p < 0.05).Conclusion: GEO treatment attenuates cisplatin-induced nephrotoxicity, in part, by modulating some inflammatory cytokines.Keywords: Zingiber officinale, Ginger, Roscoe, Essential oil, Nephrotoxicity, Cisplati

Otimização da extração de ácidos nucleicos de material de punção aspirativa por agulha fina de tiroide obtido de lâminas coradas, tecidos fixados em formalina e emblocados em parafina e amostras de sangue estocadas por longo período

OBJECTIVE: Adequate isolation of nucleic acids from peripheral blood, fine-needle aspiration cells in stained slides, and fresh and formalin-fixed/paraffin-embedded tissues is crucial to ensure the success of molecular endocrinology techniques, especially when samples are stored for long periods, or when no other samples can be collected from patients who are lost to follow-up. Here, we evaluate several procedures to improve current methodologies for DNA (salting-out) and RNA isolation. MATERIALS AND METHODS: We used proteinase K treatment, heat shock, and other adaptations to increase the amount and quality of the material retrieved from the samples. RESULTS: We successfully isolated DNA and RNA from the samples described above, and this material was suitable for PCR, methylation profiling, real-time PCR and DNA sequencing. CONCLUSION: The techniques herein applied to isolate nucleic acids allowed further reliable molecular analyses. Arq Bras Endocrinol Metab. 2012;56(9):618-26OBJETIVO: O isolamento adequado de ácidos nucleicos a partir de sangue periférico, lâmina corada de punção aspirativa por agulha fina, tecido fixado em formalina e emblocado em parafina e tecido fresco é fundamental para assegurar o sucesso de técnicas aplicadas em endocrinologia molecular, principalmente quando lidamos com amostras estocadas por longos períodos ou quando há impossibilidade de nova coleta de amostra de pacientes que perderam o seguimento. Neste trabalho, objetivamos otimizar as metodologias clássicas para a extração de DNA (salting-out) e RNA. MATERIAIS E MÉTODOS: Utilizamos proteinase K, choque térmico, dentre outras modificações, com o objetivo de aumentar a quantidade e a qualidade do material recuperado a partir das amostras descritas acima. RESULTADOS: Isolamos com sucesso DNA e RNA de tais amostras e o material obtido foi adequado para a realização de PCR, perfil de metilação, PCR em tempo real e sequenciamento de DNA. CONCLUSÃO: As técnicas aplicadas neste estudo para isolar ácidos nucleicos permitiram a realização posterior de análises moleculares consistentes e confiáveis. Arq Bras Endocrinol Metab. 2012;56(9):618-26Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal de São Paulo (UNIFESP) Escola Paulista de MedicinaFaculdade de Medicina do ABC Department of Morphology and PhysiologyUNIFESP, EPMSciEL

Chronic supplementation of beta-hydroxy-beta methylbutyrate (HM beta) increases the activity of the GH/IGF-I axis and induces hyperinsulinemia in rats

Objective: Beta-hydroxy-beta-methylbutyrate (HM beta) is a metabolite of leucine widely used for improving sports performance. Although limp is recognized to promote anabolic or anti-catabolic effects on protein metabolism, the impact of its long-term use on skeletal muscle and/or genes that control the skeletal protein balance is not fully known. This study aimed to investigate whether chronic HM beta treatment affects the activity of GH/IGF-I axis and skeletal muscle IGF-I and myostatin mRNA expression. Design: Rats were treated with HK beta (320 mg/kg BW) or vehicle, by gavage, for 4 weeks, and killed by decapitation. Blood was collected for evaluation of serum insulin, glucose and IGF-I concentrations. Samples of pituitary, liver, extensor digitorum longus (EDL) and soleus muscles were collected for total RNA or protein extraction to evaluate the expression of pituitary growth hormone (GH) gene (mRNA and protein), hepatic insulin-like growth factor I (IGF-I) mRNA, skeletal muscle IGF-I and myostatin mRNA by Northern blotting/real time-PCR, or Western blotting. Results: Chronic HM beta treatment increased the content of pituitary GH mRNA and GH, hepatic IGF-I mRNA and serum IGF-I concentration. No changes were detected on skeletal muscle IGF-I and myostatin mRNA expression. However, the HIM-treated rats although normoglycemic, exhibited hyperinsulinemia. Conclusions: The data presented herein extend the body of evidence on the potential role of HM beta-treatment in stimulating GH/IGF-I axis activity. In spite of this effect, HM beta supplementation also induces an apparent insulin resistance state which might limit the beneficial aspects of the former results, at least in rats under normal nutritional status and health conditions. (C) 2010 Growth Hormone Research Society. Published by Elsevier Ltd. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Pesquisa e Desenvolvimento (CNPq), BrazilConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CNP

Different configurations of specific thyroid hormone response elements mediate opposite effects of thyroid hormone and GC-1 on gene expression

T3 regulates transcription of the rat sarcoendoplasmic reticulum calcium ATPase in the heart. The T3 effect is mediated by three differently configured T3 response elements (TREs). Here we report the mutation of each individual TRE in the promoter and the contribution of each TRE on gene expression. Mutation of TRE1, a direct repeat element, exerted the strongest T3 response, compared with TRE2 and TRE3, which are inverted palindromes. The isolated TRE2 and TRE3, which showed no response (TRE2) or were weakly positive with T3 (TRE3), became strong negative regulatory elements with the T3 analog GC-1. We found that TRE1 recruits corepressor complexes containing nuclear receptor corepressor and histone deacetylase 3 in the absence of ligand, and steroid receptor coactivator-1-containing coactivator complexes with both T3 and GC-1. TRE3 bound the same corepressor complexes without ligand but showed only a weak association with steroid receptor coactivator-1 with T3 and a strong association with corepressor complexes with GC-1. Thus, GC-1 appears to control cofactor association differentially on these two sarcoendoplasmic reticulum calcium ATPase TREs, which could be the mechanism of ligand-dependent transcriptional activation and repression observed with the isolated TRE1 and TRE3 elements. Because the x-ray crystal structures of GC-1 and T3 complexed with the TR ligand binding domain are superimposable, the results imply that GC-1 and T3 induce differential effects on the receptor that are not evident in the static structures but must occur in the dynamic setting of receptor function. These results have implications for selective modulation of receptor function by agonist ligands

T3 acutely increases GH mRNA translation rate and GH secretion in hypothyroid rats

Cytoskeleton controls the stability of transcripts, by mechanisms that involve mRNAs and eEF1A attachment to it. Besides, it plays a key role in protein synthesis and secretion, which seems to be impaired in somatotrophs of hypothyroid rats, whose cytoskeleton is disarranged. This study investigated the: eEF1A and GH mRNA binding to cytoskeleton plus GH mRNA translation rate and GH secretion, in sham-operated and thyroidectomized rats treated with T3 or saline, and killed 30 min thereafter. Thyroidectomy reduced: (a) pituitary F-actin content, and eEF1A plus GH mRNA binding to it; (b) GH mRNA recruitment to polysome; and (c) liver IGF-1 mRNA expression, indicating that GH mRNA stability and translation rate, as well as GH secretion were impaired. T3 acutely reversed all these changes, which points toward a nongenomic action of T3 on cytoskeleton rearrangement, which might contribute to the increase on GH mRNA translation rate and GH secretion. (C) 2009 Elsevier Ireland Ltd. All rights reserved.FAPESP Fundacao de Amparo a Pesquisa do Estado de Sao Paulo[06/61713-0]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Pesquisa a Desenvolvimento (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CNPqConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESPFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Ginger Essential Oil Ameliorates Cisplatin-Induced Nephrotoxicity in Mice

Purpose: To investigate the effect of ginger essential oil (GEO) in an experimental model of cisplatininduced nephrotoxicity.Methods: Male mice were divided into treatment six groups (n = 7), namely: Groups I (saline), II and III (cisplatin, 10 mg/kg, i.p.) euthanized in 3th and 6th days, respectively, and IV, V and IV (GEO, 100, 200 and 400 mg/kg/day, respectively, by gavage 3, 4, 5 and 6 days after cisplatin injection). Creatinine levels and protein/creatinine ratio were determined in plasma and urine, respectively. Bone morphogenic protein (BMP-7) and tumor necrosis factor (TNF-α) levels of kidney tissues were determined while mRNA expression levels were obtained using real-time polymerase chain reaction.Results: GEO treatment reduced significantly creatinine levels to 0.53 ± 0.02; 0.48 ± 0.008 and 0.46 ± 0.02 at 100, 200 and 400 mg/kg, respectively, compared with control (0.70 ± 0.01) [p<0.05] but increased protein : creatinine ratio to 0.21 ± 0.01, 0.22 ± 0.01, 0.24 ± 0.02 compared with control (0.06 ± 0.008) [p<0.05]. Pro-inflammatory TNF-α mRNA expression was decreased to 1.46 ± 0.21, 1.39 ± 0.19 and 1.36 ± 0.09, at GEO doses of 100, 200 and 400 mg/kg, respectively, while anti-fibrotic BMP-7 mRNA expression increased to 2.05 ± 0.26 and 2.44 ± 0.42 at doses of 200 and 400 mg/kg, respectively, compared with control (0.59 ± 0.39, p < 0.05).Conclusion: GEO treatment attenuates cisplatin-induced nephrotoxicity, in part, by modulating some inflammatory cytokines.Keywords: Zingiber officinale, Ginger, Roscoe, Essential oil, Nephrotoxicity, Cisplati

The Effects Of Rapamycin In The Progression Of Renal Fibrosis

Renal fibrosis is a hallmark of end-stage renal diseases and of chronic allograft nephropathy (CAN). Rapamycin, besides its action through blockade of lymphocyte proliferation, also has antiproliferative, antiviral, and antitumor actions. Its use in clinical in patients with CAN has recently been advocated. Objectives: Our goal was to evaluate the effect of rapamycin in an established model of renal fibrosis, unilateral ureteral obstruction. Materials and Methods: C57BL/6 mice were divided into two groups, treated or not with daily doses of rapamycin (0.2 mg/kg) beginning on day-1. The obstruction was performed as day 0. Blood and kidney tissues were collected at 1, 4, 7, and 14 days after the surgery to quantify bone morphogenic protein (BMP)-7 and transforming growth factor (TGF)-β mRNA by real time PCR. Results: Daily treatment with rapamycin caused a significant reduction in serum creatinine at day 1 (0.57 ± 0.03 vs 0.95 ± 0.15 mg/dL, P = .002) and at day 14 (0.56 ± 0.04 vs 0.73 ± 0.07 mg/dL, P = .040). This profile was corroborated by histological morphometric analyses showing less fibrosis at day 14. However, rapamycin surprisingly induced an upregulation of TGF-β at day 4 (3.05 ± 0.46 vs 1.85 ± 0.41, P = .006) and at day 7 (6.33 ± 0.55 vs 4.97 ± 0.38, P = .024) with a reduced expression by day 14 (4.03 ± 1.07 vs 7.89 ± 0.83, P < .001). Surprisingly, rapamycin also promoted an increment in BMP-7, completely reversing the ratio of TGF-β to BMP-7, allowing a more protective phenotype. Conclusion: Rapamycin slightly ameliorated the renal dysfunction and, at later time points, induced less fibrosis and less decrease in the TGF-β to BMP-7 ratio. © 2007 Elsevier Inc. All rights reserved.392457459Wolfe, R.A., Ashby, V.B., Milford, E.L., Comparison of mortality in all patients on dialysis, patients on dialysis awaiting transplantation, and recipients of a first cadaveric transplant (1999) N Engl J Med, 341, p. 1725Hariharan, S., Johnson, C.P., Bresnahan, B.A., Improved graft survival after renal transplantation in the United States, 1988 to 1996 (2000) N Engl J Med, 342, p. 605Halloran, P.F., Melk, A., Barth, C., Rethinking chronic allograft nephropathy: the concept of accelerated senescence (1999) J Am Soc Nephrol, 10, p. 167Zeisberg, M., Hanai, J., Sugimoto, H., BMP-7 counteracts TGF-beta1-induced epithelial-to-mesenchymal transition and reverses chronic renal injury (2003) Nat Med, 97, p. 964Yang, J., Dai, C., Liu, Y., A novel mechanism by which hepatocyte growth factor blocks tubular epithelial to mesenchymal transition (2005) J Am Soc Nephrol, 161, p. 68Okada, H., Kalluri, R., Recapitulation of kidney development paradigms by BMP-7 reverses chronic renal injury (2005) Clin Exp Nephrol, 92, p. 100Stallone, G., Schena, A., Infante, B., Sirolimus for Kaposi's sarcoma in renal-transplant recipients (2005) N Engl J Med, 35213, p. 1317Lee, V.W., Chapman, J.R., Sirolimus: its role in nephrology (2005) Nephrology (Carlton), 106, p. 606Morales, J.M., Immunosuppressive treatment and progression of histologic lesions in kidney allografts (2005) Kidney Int, 99 (SUPPL), pp. S124Bumbea, V., Kamar, N., Ribes, D., Long-term results in renal transplant patients with allograft dysfunction after switching from calcineurin inhibitors to sirolimus (2005) Nephrol Dial Transplant, 2011, p. 2517Shihab, F.S., Bennett, W.M., Yi, H., Sirolimus increases transforming growth factor-beta1 expression and potentiates chronic cyclosporine nephrotoxicity (2004) Kidney Int, 654, p. 1262Song, K., Wang, H., Krebs, T.L., Novel roles of Akt and mTOR in suppressing TGF-beta/ALK5-mediated Smad3 activation (2006) EMBO J, 251, p. 58van der Poel, H.G., Hanrahan, C., Zhong, H., Rapamycin induces Smad activity in prostate cancer cell lines (2003) Urol Res, 306, p. 38