RESUMEN
O milho (Zea mays L.) possui nos estigmas (cabelo de milho) substâncias que o tornam diurético, podendo ser importante no controle da hipertensão. No presente estudo, foi investigado o efeito do extrato aquoso (EA) do cabelo de milho sobre o fluxo renal de água (V) e eletrólitos e a pressão arterial (PA) em ratos Wistar anestesiados. Foram realizados 3 grupos: I) Controle - administração intragástrica (AI) de 1mL de água destilada; II) AI de 1 mL de EA de cabelo de milho a 20 por cento e III) AI de 1 mL de solução contendo furosemida. Canulou-se a artéria carótida esquerda para mensuração da PA, de 10 em 10 minutos, e a bexiga urinária, para mensuração de V de 30 em 30 minutos e da carga excretada dos íons sódio (Qe (Na+)) e potássio (Qe (K+)). Protocolo experimental: quatro períodos de 30 minutos cada: basal (avaliação dos parâmetros basais) e experimentais (Ex) 1, 2 e 3 (30, 60 e 90 minutos após a AI, respectivamente). O Grupo I não apresentou alterações significativas entre os períodos nos parâmetros analisados (p>0,05). O Grupo II apresentou aumento significativo (p<0,05) em V, em Qe (Na+) e em Qe (K+) nos períodos Ex2 e Ex3, com redução significativa na PA (p<0,05) em Ex2 e Ex3. Conforme esperado, o Grupo III apresentou aumento significativo em V nos períodos Ex2 (p<0,05) e Ex3 (p<0,001), aumento em Qe (Na+) em Ex1 (p<0,05), Ex2 (p<0,001) e Ex3 (p<0,001) e aumento em Qe (K+) em Ex2 (p<0,05) e Ex3 (p<0,001), com redução significativa na PA (p<0,05) em Ex2 e Ex3. Os dados mostram que o EA do cabelo de milho possui efeito diurético, porém não age como um diurético "de alça", uma vez que não levou à expoliação de potássio e nem a uma excreção tão acentuada de sódio quanto à furosemida.
The corn (Z. mays) has in its stigmas (corn silks) substances that make it diuretic, which may be important in hypertension control. In this study, the effect of aqueous extract (AE) of corn silks on the renal flow of water (V) and electrolytes and arterial pressure (AP) was investigated in anesthetized Wistar rats. Three groups were tested: I) Control - intragastric administration (IA) of 1mL of distilled water, II) IA of 1 mL of AE of corn silks at 20 percent and III) IA of 1 mL of a solution containing furosemide. Cannulation was performed in the left carotid artery to measure AP, at every 10 minutes, and in the urinary bladder to measure V, at every 30 minutes, and the excreted load of ions sodium (Qe (Na+)) and potassium (Qe (K+)). Experimental protocol: four periods of 30 minutes each: basal (evaluation of basal parameters) and experimental (Ex) 1, 2 and 3 (30, 60 and 90 minutes after IA, respectively). Group I had no significant differences between periods for the analyzed parameters (p>0.05). Group II presented a significant increase (p<0.05) in V, Qe (Na+) and Qe (K+) in periods Ex2 and Ex3, with significant reduction in AP (p<0.05) in Ex2 and Ex3. As expected, Group III had a significant increase in V in periods Ex2 (p<0.05) and Ex3 (p<0.001), an increase in Qe (Na+) in Ex1 (p<0.05), Ex2 (p<0.001) and Ex3 (p<0.001) and an increase in Qe (K+) in Ex2 (p<0.05) and Ex3 (p<0.001), with an important reduction in AP (p<0.05) in Ex2 and Ex3. These data show that AE of corn silks has a diuretic effect but does not act as a loop diuretic since it did not lead to potassium loss or marked sodium loss, compared to furosemide.
Asunto(s)
Animales , Masculino , Ratas , Agua Corporal/química , Electrólitos/química , Extractos Vegetales/uso terapéutico , Presión Arterial , Ratas Wistar , Orina , Zea mays/efectos adversos , Zea mays , Diuréticos/efectos adversos , Diuréticos/química , Diuréticos/uso terapéutico , Plantas Medicinales/efectos adversos , Plantas Medicinales/toxicidadRESUMEN
Nuclear magnetic resonance is an important tool for high-resolution structural studies of proteins. It demands high protein concentration and high purity; however, the expression of proteins at high levels often leads to protein aggregation and the protein purification step can correspond to a high percentage of the overall time in the structural determination process. In the present article we show that the step of sample optimization can be simplified by selective labeling the heterologous protein expressed in Escherichia coli by the use of rifampicin. Yeast thioredoxin and a coix transcription factor Opaque 2 leucine zipper (LZ) were used to show the effectiveness of the protocol. The (1)H/(15)N heteronuclear correlation two-dimensional NMR spectrum (HMQC) of the selective (15)N-labeled thioredoxin without any purification is remarkably similar to the spectrum of the purified protein. The method has high yields and a good (1)H/(15)N HMQC spectrum can be obtained with 50 ml of M9 growth medium. Opaque 2 LZ, a difficult protein due to the lower expression level and high hydrophobicity, was also probed. The (15)N-edited spectrum of Opaque 2 LZ showed only the resonances of the protein of heterologous expression (Opaque 2 LZ) while the (1)H spectrum shows several other resonances from other proteins of the cell lysate. The demand for a fast methodology for structural determination is increasing with the advent of genome/proteome projects. Selective labeling the heterologous protein can speed up NMR structural studies as well as NMR-based drug screening. This methodology is especially effective for difficult proteins such as hydrophobic transcription factors, membrane proteins, and others.