Evaluation of potential hepatotoxicity induced by Bortezomib

Bortezomib, an inhibitor of 26S proteasome, is an anti-cancer therapeutic agent used in different cancer types. It leads to the arrest of the cancerous cell cycle by inhibiting angiogenesis and inducing apoptosis. Liver is the vital organ for detoxification and excretion of toxic products. The treatment with chemotherapy is a challenge, drugs are used to destroy cancer cells, but healthy cells can be affected during cancer treatment as well. The main objective of this study was to analyze the histopathological and biochemical effects of bortezomib on liver. Twenty-four female C57BL/6 mice were distributed into 4 groups, bortezomib injected treatment groups (Btz1, Btz2) and saline injected control groups (C1, C2). Bortezomib and saline were treated twice per week for 6 weeks and sacrificed at the end of one day (Btz1, C1) and 4 weeks (Btz2, C2) after the last injection. Liver samples were examined for histopathological analysis and the serum samples processed for biochemical analysis. Tissue samples were fixed, routinely processed, sectioned, and stained with Hematoxylin and Eosin (H&E). Periodic Acid-Schiff (PAS), Sudan Black staining and Masson's trichrome histochemical staining methods were performed to characterize the lesions. Histopathological analysis of the Btz1 and Btz2 groups revealed acute hepatic morphological changes such as hepatocellular swelling (cloudy swelling), necro-inflammatory reaction, and increased mononuclear polyploidy. Based on the negative staining with PAS and Sudan Black staining, hepatocellular swelling was diagnosed as hydropic degeneration. Necro-inflammatory reaction observed in the form of acute hepatitis was composed of mainly mononuclear cell infiltration accompanied by multifocal necrotic foci. Kupffer cell proliferation was observed in parallel with degenerative and necrotic changes. An Increase in hepatocellular mononuclear polyploidy visualized as hepatocytes with a single enlarged nucleus was detected in all liver sections of Btz1 and Btz2 groups. Individual cases of cholestasis (n = 1) and mild hepatic fibrosis (n = 1) were also reported. Significant elevated levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT) were detected in bortezomib treated groups. Few clinical cases reported liver injury related to bortezomib used for cancer treatment. However, the liver was not considered as a target for bortezomib treatment. Our data suggesting that bortezomib caused liver damage and induces elevations in serum levels. The reported hepatic lesions including hepatocellular swelling, acute hepatitis and mononuclear polyploidy were mainly mild and moderate in severity. The increase of polyploidy in liver tissue of mice treated with bortezomib in this study was explained as a reaction of the liver facing the drug-induced hepatic damage. The mechanism leading to the hepatotoxicity of bortezomib treatment is not known but the production of a toxic metabolite through its metabolism in the liver can be suggested. Moreover, no recovery was also observed in histopathological and biochemical analyses suggesting that the bortezomib effect is non-reversible four weeks after the drug was withdrawn. Patients should be informed about the possibility of acute drug-induced hepatitis and hepatotoxicity of this chemotherapeutic agent after the treatment.(AU)

Avaliação dos compostos bioativos presentes na semente de Passiflora spp. e sua influência sobre marcadores bioquímicos, oxidativos e inflamatórios de camundongos submetidos à dieta hiperlipídica / Evaluation of bioactive compounds present in Passiflora spp. seed and its influence on oxidative stress and inflammation in a high fat-fed mice

O consumo de dieta hiperlipídica e consequente acúmulo de lipídios nos adipócitos é uma condição associada ao estresse oxidativo e à perpetuação de um quadro inflamatório de leve intensidade que leva ao desenvolvimento de doenças crônicas não transmissíveis. Uma vez que alguns componentes da dieta são reconhecidos como fortalecedores do sistema antioxidante exógeno dos organismos vivos, o objetivo deste trabalho foi investigar o efeito metabólico do extrato de sementes do gênero Passiflora sobre parâmetros bioquímicos, oxidativos e inflamatórios de camundongos submetidos a uma dieta hiperlipídica. Para tanto, inicialmente realizou-se um estudo de composição química (centesimal, teores de minerais e ácidos graxos) e de otimização da extração de sementes de P. edulis Flavicarpa para obtenção de maiores teores de compostos polifenólicos com expressiva capacidade antioxidante in vitro, segundo os parâmetros de processo tempo, temperatura e concentração de etanol. Determinada a condição ideal de extração, esta foi empregada para as demais espécies em estudo P. alata BRS Mel do Cerrado e BRS Doce Mel, P. tenuifila BRS Vita e P. edulis BRS Sol do Cerrado e BRS Gigante Amarelo e P. setacea BRS Pérola do Cerrado e foi realizada o estudo de composição química. Os extratos etanólicos obtidos possuíram interessante atividade antioxidante, com destaque para a espécie P. setacea. O composto piceatanol foi o polifenol majoritário (0,41-10,28 g/ 100 g de semente em base seca) nas sementes de Passifloras analisadas, com exceção para a amostra P. setácea BRS Vita, cujo composto de íon molecular m/z 747,2 não foi identificado. As sementes ainda apresentaram alto teor de óleo, com o ácido linoleico em sua composição, e proteína. Os extratos das sementes de P. setacea BRS Pérola do Cerrado e Passiflora edulis Flavicarpa, nas concentrações de 500 e 1000 mg/Kg de ração foram utilizados para a realização do ensaio biológico. O consumo dos extratos, dependendo da dose, apresentou efeitos biológicos importantes, tais como a diminuição das concentrações séricas de colesterol, glicose, insulina e leptina a níveis aproximados ao determinado para os animais em consumo de dieta normolipídica. Adicionalmente, verificou-se a atenuação do estresse oxidativo hepático, por elevação da atividade enzimática das enzimas catalase e glutationa peroxidase e diminuição da lipoperoxidação, e do processo inflamatório, por redução da concentração tecidual das citocinas IL-6 e MCP-1. Os resultados obtidos neste estudo contribuem para o conhecimento a respeito das passifloras brasileiras e na potencial utilização das sementes, consideradas subprodutos, na contribuição da manutenção da saúde

The consumption of a high fat diet and consequent excessive lipid accumulation in adipocytes is a condition associated with oxidative stress and perpetuation of a mild inflammatory condition that leads to the development of chronic diseases. Since some dietary components are recognized as empowering exogenous antioxidant system defenses of living organisms, the aim of this study is to investigate the metabolic effects of passion fruit seeds that possess a high content of bioactive compounds and high antioxidant capacity in vitro on oxidative stress and inflammatory conditions in mice subjected to a high fat diet. For this purpose, initially were performed a chemical composition study (proximate, minerals and fatty acids content) and extraction optimization of P. edulis Flavicarpa seed for obtaining higher levels of polyphenolic compounds with expressive antioxidant capacity in vitro through process parameters such as time temperature and ethanol concentration. Once the optimum condition of extraction was determined, it was applied to others studied species P. alata BRS Mel do Cerrado e BRS Doce Mel, P. tenuifila BRS Vita e P. edulis BRS Sol do Cerrado e BRS Gigante Amarelo e P. setacea BRS Pérola do Cerrado and the chemical composition study was carried out. The obtained ethanolic extracts had high antioxidant activity, particularly the species P. setacea. The piceatannol compound was the major polyphenol (0.41 to 10.28 g / 100 g seed in dry basis) in the analyzed Passiflora, except for the sample P. setacea BRS Vita, which molecular íon m/z 747.2 was not identified. The seeds also showed high content of oil, with linoleic acid in its composition, and protein. P. setacea BRS Pérola do Cerrado and Passiflora edulis Flavicarpa seed extracts at concentrations of 500 and 1000 mg / kg of feed were used to carry out the biological assay. The consumption of the extracts, depending on the concentration, exhibited significant biological effects, such as the reduction of serum cholesterol, glucose, insulin and leptin levels to those one observed in animals with normolipidic diet consumption. Additionally, hepatic oxidative stress was attenuated by elevating enzymatic activity of catalase and glutathione peroxidase and decrease in the lipid peroxidation and inflammation by reducing the tissue concentrations of IL-6 and MCP-1 cytokines. The results obtained in this study contributes to the knowledge of the Brazilian passiflora and potential use of the seeds, considered a by-products, in health maintenance

Avaliação hematológica e bioquímica em ratas prenhes tratadas com extrato de ginkgo bilob / Hematologic and biochemistry evaluation on pregnant rat treated with gingko biloba extract

Introdução: O extrato de Gingko biloba (GBE) é um fitoterápico usado no tratamento de doenças degenerativas e em estudos recentes tem sido demonstrado efeito nefro e hepatoprotetor de seus componentes. Material e métodos: No presente estudo, 120 ratas Wistar prenhes foram distribuídas em dois grupos experimentais ­ GB 15 e GB 21 ­ tratadas, respectivamente, do primeiro ao oitavo dia da prenhez e do oitavo ao vigésimo dia com 0, 3,5; 7,0 ou 14mg/kg/dia de extrato aquoso de GBE, via gavagem. Os animais foram eutanaziados por exsanguinação total, sob anestesia, no 15º dia ou no 20º dia de prenhez. Os seguintes parâmetros foram avaliados no sangue coletado: eritrograma, leucograma, dosagens séricas de ureia, creatinina, ALT, AST, colesterol e triglicérides. Resultados: Não foram encontradas alterações significativas no padrão hematológico de ratas tratadas nos grupos GB 15 e GB 21. Em relação ao perfil bioquímico, o grupo GB 15, tratado com as doses de 7 e 14mg/kg, evidenciou aumento dos níveis de colesterol e redução de ALT, ureia e creatinina. No grupo GB 21, tratado com as mesmas doses, não se observou aumento de colesterol, mas sim de ureia, enquanto que ALT e creatinina reduziram-se da mesma maneira que no grupo GB 15. Conclusões: Os resultados sugerem que o GBE não altera os padrões hematológicos, porém, no início da gestação aumenta os níveis de colesterol, enquanto que no final da gestação não altera o colesterol, aumentando a ureia, e durante os dois períodos de gestação reduz creatinina e ALT, o que parece confirmar os efeitos nefro e hepatoprotetor.

Introduction: The Ginkgo biloba extract (GBE) is a phytotherapic used in the treatment of neurodegenerative diseases and recent studies have demonstrated nephro and hepatoprotector effects of its components. Material and methods: In this study 120 pregnant Wistar rats were distributed among two experimental groups - GB15 e GB21 ­ treated respectively from the first to eight day of pregnancy and to the eight to de 20th day, with zero, 3.5, 7 or 14mg/kg/day of aqueous extract of Gingko biloba by gavagem. Animals were euthanized by exsanguinations under anesthesia on 15th or 21th pregnancy day. The following parameters were analyzed in the blood hemogram, hematocrit, hemoglobin, total leukocytes, cholesterol, triglycerides, urea, creatinine, aspartato aminotransferase (AST/TGO), alanina aminotransferase (ALT/TGP). Results: No hematological alteration was observed in either group. With respect to biochemistry profile the GB15, group treated with 7 and 14mg/kg, showed higher level of cholesterol and lower level of ALT, urea and creatinin. In the group GB21, treated with the same dose, there was no cholesterol alteration but higher level of urea whereas ALT and creatinin where lower than control as in GB15 group. Conclusions: GBE seems do not alter hematological profile but at early gestation increase the cholesterol level. At latter gestation do not alter cholesterol but increase urea levels. At all period of the gestation the GBE decrease creatinine and ALT seems to confirm possible nepro and hepatic protector effect.

Avaliação dos parâmetros bioquímicos e hematológicos em ratas gestantes tratadas com extrato aquoso de piper methysticum: observações preliminares / Evaluation of biochemical and hematological parameters in pregnant rats treated with aqueous extract of piper methysticum: preliminary observations

Objetivo: No presente trabalho são avaliados parâmetros hematológicos e bioquímicos de ratas expostas ao extrato aquoso de Piper methysticum no período de implantação do blastocisto. Métodos: Quarenta ratas Wistar prenhes foram organizadas aleatoriamente nos grupos controle e tratado, que receberam respectivamente 1 ml de água destilada e 10 mg de extrato aquoso de kava / kg de peso corporal em 1 ml de água destilada. O tratamento foi feito por via intragástrica, duas vezes ao dia, nos dias 5, 6 e 7 pós - inseminação (1o dia=espermatozóide no esfregaço) e eutanasiadas, no 15o dia de prenhez, por exsanguinação total (punção cardíaca) sob anestesia. Resultados: houve leucopenia e aumento signifi cativo na dosagem de Transaminase Glutâmico Oxalacética (TGO) entre os animais do grupo tratado. Contudo, não foram observadas alterações histopatológicas no fígado. Conclusão: O extrato aquoso de kava-kava, administrado a ratas prenhes, parece não causar toxicidade hepática, embora tenha evidenciado leucopenia e aumento de TGO.

Aims: To evaluate biochemistry and hematological parameters in pregnant rats treated with aqueous extract of Piper methysticum. Methods: Forty pregnant rats were randomly distributed into two groups: Control that received 1ml of distilled water and treated with 10mg of aqueous extract of kava/kg/body weight, by gavage, twice a day, from day-6 to 7 post insemination (day 1 = spermatozoa visible in the vaginal smear). Euthanize was performed by exsanguinations via cardiac punction under anesthesia (ketamine plus xylazine) at 14 post insemination day. Results: Treated animal presents leucopenia and higher concentration of TGO. No histopathologic alteration was observed in the liver. Conclusion: Th e Piper methysticum aqueous extract seems do not cause hepatotoxicity, but cause leucopenia and increase the plasmatic level of TGO.