Resveratrol and resveratrol-hydroxypropyl-β-cyclodextrin complex recovered the changes of creatine kinase and Na+, K+-ATPase activities found in the spleen from streptozotocin-induced diabetic rats

Type 1 diabetes (T1D) is the result of the selective destruction of the pancreatic β-cells by T cells of the immune system. Although spleen is a secondary lymphoid organ, it is also involved in the T1D pathogenesis. However, the alterations in a variety of cellular processes of this disease need to be further understood. We aimed to analyze the benefits of resveratrol, and its complexed form on diabetic complications in the spleen of rats. To this end, we investigated important enzymes of phosphoryl transfer network, and Na+, K+-ATPase activity. Wistar rats were divided into non-diabetic groups: Control, Ethanol, Resveratrol, Hydroxypropyl-β-cyclodextrin, Resveratrol-hydroxypropyl-β-cyclodextrin, and diabetic groups with the same treatments. Diabetes was induced by a single dose of 60 mg/kg of streptozocin intraperitoneally, and treatments by intragastric gavage once daily for 60 days. Hyperglycemia reduced creatine kinase activity, which was reversed by the administration of resveratrol. Na+, K+-ATPase activity was greatly affected, but it was reversed by resveratrol and resveratrol-hydroxypropyl-β-cyclodextrin. This suggest an energetic imbalance in the spleen of diabetic rats, and in case this also occurs in the diabetic patients, it is possible that resveratrol supplementation could be beneficial to the better functioning of the spleen in diabetic patients

β-Cyclodextrins alter the energy metabolism-related enzyme activities in rats

Abstract Although widely used in medicine, separation technology, and other fields, the effects of cyclodextrins on the activities of phosphoryl transfer enzymes have not been previously evaluated. In vivo studies evaluated the function of cyclodextrins as active compounds. Despite the use of cyclodextrins as active compounds, the effects of cyclodextrins on hepatic and renal tissues remain to be fully elucidated. The primary objective of this study was to evaluate the effects of β- cyclodextrins, methyl-β-cyclodextrin (M-β- cyclodextrins), and (2-hydroxypropyl)-β-cyclodextrin (HP-β-cyclodextrins) on enzyme activities regulating the maintenance of energy homeostasis in the kidney and liver tissues in relation to toxicity. Serum levels of liver and kidney markers were measured, and oxidative stress parameters were assessed. After 60-day treatments, we observed that the administration of β-cyclodextrins and M-β-cyclodextrins inhibited the hepatic activity of pyruvate kinase, an irreversible enzyme within the glycolytic pathway. Additionally, administration of HP-β-cyclodextrins inhibited creatine kinase activity and increased the total sulfhydryl content in kidneys. Here, we demonstrated for the first time that β-cyclodextrins, M-β-cyclodextrins, and HP-β-cyclodextrins cause bioenergetic dysfunction in renal and hepatic tissues. These findings suggest that understanding the balance between cyclodextrins’ efficacy and adverse effects is essential for better accepting their use in medicine

AVALIAÇÃO DE LIPOSSOMAS COM CREATINA SOBRE O METABOLISMO CEREBRAL DE RATOS WISTAR COM HIPERFENILALANINEMIA

Submitted by MARCIA ROVADOSCHI ([email protected]) on 2018-08-16T19:41:42Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_NathanaJamilleMezzomo.pdf: 2403536 bytes, checksum: a811beb07b8f995e286c3f3f76fad6bd (MD5)Made available in DSpace on 2018-08-16T19:41:42Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_NathanaJamilleMezzomo.pdf: 2403536 bytes, checksum: a811beb07b8f995e286c3f3f76fad6bd (MD5) Previous issue date: 2015-03-27Phenylketonuria (PKU) is an inborn error of phenylalanine (Phe) metabolism caused by a deficiency of hepatic phenylalanine hydroxylase enzyme (EC 1.14.16.1 - PAH), which converts Phe tyrosine (Tyr), resulting in hyperphenylalaninemia. Excess Phe compromises the developing brain of affected individuals. Considering the creatine energy and neuroprotective substance, the aim of use it in liposomal form was to increase its concentration in the brain and evaluate parameters of energy metabolism, oxidative stress and behavioral usually changed by excess Phe in PKU patients or hyperphenylalaninemia (HPA) animals. Also, check whether the administrations of nanoparticles could affect non-PAH rats parameters, thus evaluating a neurotoxic action. The ethanol injection method for production of liposomes containing creatine allowed the formation of nanoscale particles with pH around the saline, with an average size of 236.44 nm, a polydispersity index below 0.3, mean zeta potential of -23.9 mV, encapsulation efficiency of 33.65% and creatine content of 93%. The HPA alter the activity of cytosolic and mitochondrial fractions of creatine kinase (CK) in the cerebral cortex and the hippocampus in the mitochondrial fraction, however, did not affect the activity of pyruvate kinase (PK) and adenilato kinase (AK) the analyzed brain structures. The administration of liposomes containing creatine (LCr) possibly prevented the alterations of cytosolic and mitochondrial CK activity in the cerebral cortex of the HPA group. Induction HPA did not change the oxidative stress parameters evaluated in the brain cortex and hippocampus. However, administration of LCr in animals non-HPA, increased levels of GSH antioxidant and superoxide dismutase activity (SOD) and in contrast, stimulated an increased of dihydrodichlorofluorescein oxidation (DCFH) in cerebral cortex. The blank liposomes (LBr) also increased the reduced glutathione (GSH) content in the cerebral cortex. Associated with these results, one can also check the cerebral impairment of the HPA animals through the reduced brain mass, open field test, tail suspension and elevated zero maze. The administration of LCr the HPA animals can partially prevent changes in enzyme activity, as well as the number of rearings improved open field test and the number of head-dipping of the elevated zero maze test. In short, we believe that the LCr may have crossed the blood-brain barrier, since its effects were differentiated administration of free creatine (Cr), thus preventing changes caused by Phe administration on behavioral tests and enzyme activity energy metabolism in the cortex and hippocampus.A fenilcetonúria (PKU) é um erro inato do metabolismo do aminoácido fenilalanina (Phe) provocado pela deficiência da enzima hepática fenilalanina hidroxilase (EC 1.14.16.1 - PAH), que converte Phe em tirosina (Tyr), resultando em hiperfenilalaninemia. O excesso de Phe compromete o desenvolvimento cerebral dos indivíduos afetados. Considerando a creatina uma substância energética e neuroprotetora, o objetivo de utilizá-la na forma lipossomal foi o de aumentar sua concentração no cérebro e avaliar parâmetros do metabolismo energético, do estresse oxidativo e comportamentais, normalmente alterados pelo excesso de Phe em pacientes PKU ou em animais com hiperfenilalaninemia (HPA). Além de verificar se as administrações das nanopartículas poderiam afetar os parâmetros de ratos não-HPA, avaliando dessa forma uma ação neurotóxica. O método de injeção de etanol utilizado para a produção dos lipossomas contendo creatina permitiu a formação de partículas nanométricas com pH próximo ao da salina, com tamanho médio de 236,44 nm, índice de polidispersão abaixo de 0,3, potencial zeta médio de -23,9 mV, eficiência de encapsulação de 33,65 % e teor de 93 %. A HPA alterou a atividade das frações citosólica e mitocondrial da creatinacinase (CK) no córtex cerebral e na fração mitocondrial no hipocampo, porém, não afetou a atividade da piruvatoquinase (PK) e adenilatoquinase (AK) nas estruturas cerebrais estudadas. A administração de lipossomas contendo creatina (LCr), possivelmente preveniu as alterações da atividade da CK citosólica e mitocondrial no córtex cerebral dos grupo HPA. A indução a HPA não alterou os parâmetros de estresse oxidativo avaliados no córtex cerebral e hipocampo. No entanto, a administração de LCr nos animais não-HPA, aumentou os níveis do antioxidante glutationa reduzida (GSH) e a atividade da superóxido dismutase (SOD) e em contrapartida, estimularam uma maior oxidação de diclodihidrofluoresceína (DCFH) em córtex cerebral. Os lipossomas brancos (LBr) também elevaram a concentração de GSH no córtex cerebral. Associado a esses resultados, pode-se também verificar o comprometimento cerebral dos animais HPA por meio da redução da massa cerebral, testes de campo aberto, suspensão da cauda e labirinto zero elevado. A administração de LCr nos animais HPA, pode prevenir parcialmente alterações das atividades enzimáticas, assim como melhorou o número de levantamentos do teste de campo aberto e o número de espiadas do teste labirinto zero elevado. Em suma, acreditamos que os LCr possam ter atravessado a barreira sangue-cérebro, uma vez que seus efeitos foram diferenciados da administração da creatina livre (Cr), conseguindo prevenir alterações causadas pela administração de Phe sobre o testes comportamentais e a atividade de enzimas do metabolismo energético do córtex e hipocampo

Resveratrol and resveratrol-hydroxypropyl-β-cyclodextrin complex recovered the changes of creatine kinase and Na+, K+-ATPase activities found in the spleen from streptozotocin-induced diabetic rats

Type 1 diabetes (T1D) is the result of the selective destruction of the pancreatic β-cells by T cells of the immune system. Although spleen is a secondary lymphoid organ, it is also involved in the T1D pathogenesis. However, the alterations in a variety of cellular processes of this disease need to be further understood. We aimed to analyze the benefits of resveratrol, and its complexed form on diabetic complications in the spleen of rats. To this end, we investigated important enzymes of phosphoryl transfer network, and Na+, K+-ATPase activity. Wistar rats were divided into non-diabetic groups: Control, Ethanol, Resveratrol, Hydroxypropyl-β-cyclodextrin, Resveratrol-hydroxypropyl-β-cyclodextrin, and diabetic groups with the same treatments. Diabetes was induced by a single dose of 60 mg/kg of streptozocin intraperitoneally, and treatments by intragastric gavage once daily for 60 days. Hyperglycemia reduced creatine kinase activity, which was reversed by the administration of resveratrol. Na+, K+-ATPase activity was greatly affected, but it was reversed by resveratrol and resveratrol-hydroxypropyl-β-cyclodextrin. This suggest an energetic imbalance in the spleen of diabetic rats, and in case this also occurs in the diabetic patients, it is possible that resveratrol supplementation could be beneficial to the better functioning of the spleen in diabetic patients