ABSTRACT
INTRODUCTION: In vascular diseases, the interruption of the local blood flow and the subsequent reperfusion of oxygen can cause deleterious oxidative effects on the cells. Turmeric (Curcuma longa L.) presents the capacity to neutralize free radicals along with preventive and therapeutic effects for several diseases. OBJECTIVE: To analyze the bioactive compounds and the antioxidant capacity of the ethanolic extract of Curcuma (EEC), to evaluate its effect on human umbilical vein endothelial cells, and to analyze its effect on cellular signaling pathways. METHODS: Cells were exposed to different concentrations of EEC for 24, 48, and 72 h. Folin-Ciocalteau test, HPLC-Fluorescence analysis, and DPPH method were used to determine the phenolic compounds, curcumin content, and antioxidant action, respectively; the tetrazolium salt reduction to obtain cell viability, cytotoxicity, and the concentration that inhibits 50% of cell viability; and the immunocytochemistry technique to analyze the expression of caspase3, SIRT1, and mTOR. RESULTS: We found the presence of polyphenols in the classes of phenolic acids and curcuminoids in EEC, with 16.7% curcumin content. The number of antioxidants needed to reduce the initial DPPH concentration by 50% was 18.1 µmol/g. The extract mitigated cell damage at a dosage of 100 µg/ml, decreased the immunoexpression of caspase3, and promoted the signaling of the SIRT1 and mTOR survival pathways. CONCLUSION: EEC had a protective effect on human umbilical vein endothelial cells, subjected to oxidative stress, with decreased apoptosis (caspase3) at lower concentrations, cytoprotection by maintaining essential cell functions (mTOR), and signaling of the survival pathway (SIRT1).
INTRODUÇÃO: Em doenças vasculares, a interrupção do fluxo sanguíneo locale subsequente reperfusão de oxigênio pode causar efeitos deletérios e danos irreparáveis às células. Curcuma (Curcuma longa L.) neutraliza radicais livres além de apresentar efeitos preventivos e terapêuticos. OBJETIVO: Caracterizar os compostos bioativos e a capacidade antioxidante do extrato etanólico de cúrcuma (EEC); avaliar seu efeito nas células endoteliais da veia umbilical humana, e analisar a expressão de vias de sinalização celular. MÉTODOS: As células foram expostas a diferentes concentrações de EEC por 24, 48 e 72 horas. Utilizamos o teste de Folin-Ciocalteau, análise por HPLC-Fluorescência e método DPPH para determinar os compostos fenólicos, conteúdo de curcumina e ação antioxidante, respectivamente; o método de redução de tetrazólio para viabilidade celular, a citotoxicidade e a concentração que inibe 50% da viabilidade celular; e a técnica de imunocitoquímica para analisar a expressão de caspase3, SIRT1 e mTOR. RESULTADOS: Observou-se presença de polifenóis nas classes de ácidos fenólicos e curcuminóides no EEC, com teor de curcumina de 16,7%. A quantidade de antioxidante necessária para reduzir a concentração inicial de DPPH em 50% foi de 18,1 µmol/g. O extrato mitigou o dano celular na dosagem de 100 µg/ml, diminuiu a imunoexpressão da caspase3 e promoveu a sinalização das vias de sobrevivência SIRT1 e mTOR. CONCLUSÃO: O EEC teve efeito protetor nas células endoteliais de veia umbilical humana, submetidas ao estresse oxidativo, com diminuição da apoptose (caspase3) em concentrações mais baixas, citoproteção pela manutenção das funções celulares essenciais (mTOR) e sinalização da via de sobrevivência (SIRT1).
Subject(s)
Umbilical Veins , Oxidative Stress , Curcumin , Curcuma , Endothelial Cells , Tetrazolium Salts , Immunohistochemistry , Inhibitory Concentration 50 , AntioxidantsABSTRACT
The evaluated ammonia volatilization rates and its association with humidity, temperature, and pH in broiler litter in conventional and dark house rearing systems. Evaluations were performed at 0.01, 1.00, and 1.50 m high, using a completely randomized design, at a weekly frequency for 12 weeksfrom the sanitary void to the end of the 42-day cycle. Litter temperature had no significant difference for different types of house. Humidity and pH levels varied according to houses and evaluation periods. No ammonia volatilization was detected during initial periods. However, up from the 21st day of rearing, rates started to increase, showing higher concentrations at 0.01 m height. At 1.00 and 1.50 m, values around 20 mL·m3, were obtained and considered an acceptable exposure level for the broilers and employees directly working in this sector. Yet, broiler breeders should target lower levels due to environmental impacts.
Subject(s)
Animals , Volatilization , Chickens , Inhalation Exposure/analysis , Ammonia/pharmacokinetics , Occupational Health ServicesABSTRACT
PURPOSE: To establish a model of chronic kidney disease in White New Zealand rabbits, using an exclusive unilateral technique of renal ischemia and reperfusion. METHODS: Twenty males White New Zealand rabbits were used. All animals were subjected to the following: pre-surgical blood collection (1st collection) for creatinine and urea serum analysis, left renal ischemia and reperfusion surgery technique, another blood sample was collected after 6 weeks post surgery (2nd collection), the last blood sample (3rd collection) blood sample was taken 11 weeks post surgery (pre-euthanasia), euthanasia and withdrawal of right and left kidney for histopathological analysis. RESULTS: The creatinine levels after surgery was statistically significant higher in the 3rd collection, regarding the 1st and 2nd collection (p<0.05). A significant statistic increase for urea was showed only in the 2nd collection (p<0.05) when compared to the 1st and 3rd collections. Histopathological analysis showed bilateral lesions in the renal tissue, consistent to the process of ischemia and reperfusion. CONCLUSION: This exclusive unilateral technique of renal ischemia and reperfusion without nephrectomy in White New Zealand rabbits, showed effectiveness in getting an animal model of chronic kidney disease .