Determinación química de Hemoglobina Glicosilada

La hemoglobina [Hb] es modificada en el organismo al reaccionar con la glucosa formando la hemoglobina glicosilada. Su determinación permite reducir al máximo ciertos trastornos que se manifiestan en los períodos terminales de la diabetes. La Hb, es una proteína conjugada con una porción proteica y un grupo prostético, en los humanos la Hb es de cuatro tipos. Alfa, beta, gama y delta, cromatográficamente se han descubierto otros tipos originados por modificaciones post-traslacionales, este grupo es llamado grupo de las hemoglobinas rápidas. La hiperglucemia permanente es la que produce la glicosilación, en este proceso la glucosa se une a las proteínas sin participación de enzimas si no por enlaces covalentesDoctor en Bioquímica y FarmaciaCuenc

PLoS ONE

Over the past decade, zebrafish (Danio rerio) have emerged as an attractive model for in vivo drug discovery. In this study, we explore the suitability of zebrafish larvae to rapidly evaluate the anti-inflammatory activity of natural products (NPs) and medicinal plants used in traditional medicine for the treatment of inflammatory disorders. First, we optimized a zebrafish assay for leukocyte migration. Inflammation was induced in four days post-fertilization (dpf) zebrafish larvae by tail transection and co-incubation with bacterial lipopolysaccharides (LPS), resulting in a robust recruitment of leukocytes to the zone of injury. Migrating zebrafish leukocytes were detected in situ by myeloperoxidase (MPO) staining, and antiinflammatory activity was semi-quantitatively scored using a standardized scale of relative leukocyte migration (RLM). Pharmacological validation of this optimized assay was performed with a panel of anti-inflammatory drugs, demonstrating a concentration-responsive inhibition of leukocyte migration for both steroidal and non-steroidal anti-inflammatory drugs (SAIDs and NSAIDs). Subsequently, we evaluated the bioactivity of structurally diverse NPs with well-documented antiinflammatory properties. Finally, we further used this zebrafish-based assay to quantify the anti-inflammatory activity in the aqueous and methanolic extracts of several medicinal plants. Our results indicate the suitability of this LPS-enhanced leukocyte migration assay in zebrafish larvae as a front-line screening platform in NP discovery, including for the bioassayguided isolation of anti-inflammatory secondary metabolites from complex NP extracts.Cuencavolumen 8; número 1

LPS-enhanced leukocyte migration assay in 4 dpf zebrafish larvae.

<p>All larvae (<i>nacre</i>) are four days post-fertilization (4 dpf), with anterior to the left, scale bar = 10 µm. <b>A</b>, tail of alive larva without tail cut; <b>B</b>, tail of alive larva with tail cut; <b>C–D</b>, whole-mount MPO staining in uncut tails of zebrafish larvae; <b>C</b>, without lipopolysaccharides (−LPS) and <b>D</b>, with lipopolysaccharides (+LPS); <b>E–F</b>, whole-mount MPO staining in cut tails of zebrafish larvae; <b>E,</b> without the inclusion of LPS; <b>F</b>, with the inclusion of LPS. Dark-spots (marked by arrows) represent the migrating leukocytes, which are semi-quantified in the region to the right of the dashed red arc.</p

Anti-inflammatory activity of natural products.

a<p>Leukocyte migration is expressed as relative values (RLM).</p>***<p>p<0.001,</p>**<p>0.001*</p><p>0.01</p

Inhibitory effects of some natural products in various inflammatory targets.

<p>Inhibitory effects of some natural products in various inflammatory targets.</p

Scoring of leukocyte migration in the transected tails of zebrafish larvae.

<p>All larvae (<i>nacre</i>) are four days post-fertilization (4 dpf), with anterior to the left, scale bar = 10 µm. Dark spots (marked by arrows) represent leukocytes migrating to the injured zone in the transected tails. Migrating leukocytes were counted in the region to the right of the dashed red arc. <b>A</b>, tail of an uncut larva; <b>B</b>, tail-cut with score 0; <b>C</b>, tail-cut with score 1; <b>D</b>, tail-cut with score 2; <b>E</b> tail-cut with score 3; <b>F,</b> tail-cut with score 4. <b>^a</b>Experimental values obtained for each larvae are normalized to a relative value expressed as relative leukocyte migration (RLM); +LPS: with inclusion of lipopolysaccharides. <b>^b</b>Percentage of anti-inflammatory activity is obtained as (1−RLM)x100.</p

Inhibition of the leukocyte migration by known anti-inflammatory drugs and non anti-inflammatory compounds.

<p>After exposing the larvae to mechanical and biological damages, anti-inflammatory drugs and non anti-inflammatory compounds were evaluated for their activity in the LPS-enhanced leukocyte migration assay. Migrating MPO-positive cells were counted in the tail tip and the results expressed as relative leukocyte migration (RLM) values. Results for NSAIDs (upper panel) and SAIDs (middle panel) show a significant inhibition of leukocytes migration in a concentration-dependent manner while non anti-inflammatory drugs (lower panel) indicate no significant effect on leukocyte migration. In each case, values are plotted as RLM (SEM, n = 3 replicates with 10 larvae each) and a value of 0.5 was established as a cut-off for anti-inflammatory activity. ***p<0.001, **0.001</p

Anticonvulsant activity of bisabolene sesquiterpenoids of Curcuma longa in zebrafish and mouse seizure models.

urmeric, obtained from the rhizomes of Curcuma longa, is used in South Asia as a traditional medicine for the treatment of epilepsy. To date, in vivo studies on the anticonvulsant activity of turmeric have focused on its principal curcuminoid, curcumin. However, poor absorption and rapid metabolism have limited the therapeutic application of curcumin in humans. To explore the therapeutic potential of turmeric for epilepsy further, we analyzed its anticonvulsant activity in a larval zebrafish seizure assay. Initial experiments revealed that the anticonvulsant activity of turmeric in zebrafish larvae cannot be explained solely by the effects of curcumin. Zebrafish bioassay-guided fractionation of turmeric identified bisabolene sesquiterpenoids as additional anticonvulsants that inhibit PTZ-induced seizures in both zebrafish and mice. Here, we present the first report of the anticonvulsant properties of bisabolene sesquiterpenoids and provide evidence which warrants further investigation toward the mechanistic understanding of their neuromodulatory activity