RESUMO
Bumelia sartorum (Sapotaceae) is used ethnomedicinally for treatment of several diseases, including diabetes mellitus. The aim of this work was to investigate the hypoglycemic effect of B. sartorum extracts, rich in polyphenolic compounds, and the possible mechanisms of action. Assessment of B. sartorum hypoglycemic activity was performed from the blood glucose level in normoglycemic mice after administration of the extract by oral gavage. The hypothesis that sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibition could prolong the increase in cytoplasmic Ca2+ concentration, thus leading to an increase of insulin release was evaluated. The enzyme inhibition was measured by ATP hydrolysis using SERCA1 isolated from rabbit skeletal muscle. The total content of phenolic compounds was determined by the Folin-Ciocalteau method. The ethyl acetate (EtOAc) partition and F5 fraction obtained from B. sartorum, both of them rich in polyphenolics, were shown to have a hypoglycemic effect on normoglycemic mice, more significant than that of the known antidiabetic drug, glibenclamide used as a standard comparable compound. Both samples significantly inhibited SERCA activity. Different extracts of B. sartorum, rich in polyphenolic compounds, were able to reduce blood glucose in normoglycemic mice and inhibit SERCA activity. SERCA inhibition may be one of the possible mechanisms involved in glucose decrease.
Assuntos
Hipoglicemiantes/farmacologia , Extratos Vegetais/farmacologia , Polifenóis/farmacologia , Sapotaceae/química , Animais , Glicemia/análise , Cálcio/metabolismo , Feminino , Camundongos , Coelhos , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/antagonistas & inibidoresRESUMO
Excessive activation of NMDA glutamate receptors and the resulting loss of intracellular Ca(2+) homeostasis may be lethal (excitotoxic) to neurons. Such excitotoxicity can be induced in vivo by intrastriatal infusion of quinolinate, as this substance selectively activates NMDA receptors. The aim of the present research was to investigate whether the in vivo treatment of striatal tissue with quinolinate would lead to an early impairment of sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) activity or mitochondrial Ca(2+) sequestration, two intracellular mechanisms involved in Ca(2+) homeostasis and signaling. Sodium quinolinate was infused intrastriatally into adult rats, and 6 h later the brains were removed and the corpora striata dissected. At this time point, striatal sections stained with Fluoro-Jade, a cellular marker of cell death, showed initial signs of neuronal degeneration. In addition, SERCA activity decreased 39% in relation to the activity observed in the control striata. A corresponding decrease of the same magnitude in (45)Ca(2+) uptake by striatal microsomes was also found in the treated striata. Western blot analysis did not indicate any decrease in SERCA levels in striatal tissue after quinolinate infusion. Mitochondrial Ca(2+) sequestration was still preserved in quinolinate-treated striatal tissue when the assay was carried out in the presence of physiological concentrations of ATP and Mg(2+). These results suggest that impairment of the SERCA function may be an early event in excitotoxicity.
Assuntos
Corpo Estriado/efeitos dos fármacos , Retículo Endoplasmático/enzimologia , Agonistas de Aminoácidos Excitatórios/farmacologia , Neurotoxinas/farmacologia , Ácido Quinolínico/farmacologia , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/metabolismo , Animais , Cálcio/metabolismo , Feminino , Homeostase , Mitocôndrias/metabolismo , Ratos , Ratos Wistar , Receptores de N-Metil-D-Aspartato/metabolismoRESUMO
Previous data from our laboratory showed that the reticulum of the sea cucumber smooth muscle body wall retains both a sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) and a sulfated polysaccharide. In this invertebrate, the transport of Ca(2+) by the SERCA is naturally inhibited by these endogenous sulfated polysaccharides. The inhibition is reverted by K(+) leading to an enhancement of the Ca(2+) transport rate. We now show that vesicles derived from the endoplasmic reticulum of unfertilized eggs from the sea urchin Arbacia lixula retain a SERCA that is able to transport Ca(2+) at the expense of ATP hydrolysis. As described for the sea cucumber SERCA isoform, the enzyme from the sea urchin is activated by K(+) but not by Li(+) and is inhibited by thapsigargin, a specific inhibitor of SERCA. A new sulfated polysaccharide was identified in the sea urchin eggs reticulum composed mainly by galactose, glucose, hexosamine and manose. After extraction and purification, this sulfated polysaccharide was able to inhibit the mammal SERCA isoform found in rabbit skeletal muscle and the inhibition is reversed by K(+). These data suggest that the regulation of the SERCA pump by K(+) and sulfated polysaccharides is not restricted to few marine invertebrates but is widespread.
Assuntos
Óvulo/metabolismo , Polissacarídeos/fisiologia , Potássio/farmacologia , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/metabolismo , Animais , Cálcio/metabolismo , Indóis/farmacologia , Cloreto de Lítio/farmacologia , Músculos/efeitos dos fármacos , Músculos/metabolismo , Oxalatos/farmacologia , Fosfatos/farmacologia , Coelhos , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/antagonistas & inibidores , Ouriços-do-Mar , Ésteres do Ácido Sulfúrico/farmacologia , Tapsigargina/farmacologiaRESUMO
We report for the first time that marine angiosperms (seagrasses) possess sulfated polysaccharides, which are absent in terrestrial and freshwater plants. The structure of the sulfated polysaccharide from the seagrass Ruppia maritima was determined. It is a sulfated D-galactan composed of the following regular tetrasaccharide repeating unit: [3-beta-D-Gal-2(OSO3)-1-->4-alpha-D-Gal-1-->4-alpha-D-Gal-1-->3-beta-D-Gal-4(OSO3)-1-->]. Sulfated galactans have been described previously in red algae and in marine invertebrates (ascidians and sea urchins). The sulfated galactan from the marine angiosperm has an intermediate structure when compared with the polysaccharides from these two other groups of organisms. Like marine invertebrate galactan, it expresses a regular repeating unit with a homogenous sulfation pattern. However, seagrass galactan contains the D-enantiomer of galactose instead of the L-isomer found in marine invertebrates. Like red algae, the marine angiosperm polysaccharide contains both alpha and beta units of D-galactose; however, these units are not distributed in an alternating order, as in algal galactan. Sulfated galactan is localized in the plant cell walls, mostly in rhizomes and roots, indicative of a relationship with the absorption of nutrients and of a possible structural function. The occurrence of sulfated galactans in marine organisms may be the result of physiological adaptations, which are not correlated with phylogenetic proximity. We suggest that convergent adaptation, due to environment pressure, may explain the occurrence of sulfated galactans in many marine organisms.
Assuntos
Alismatales/química , Evolução Biológica , Galactanos/análise , Galactanos/química , Hydrocharitaceae/química , Enxofre/química , Sequência de Carboidratos , Parede Celular/química , Galactanos/metabolismo , Transferência Genética Horizontal , Espectroscopia de Ressonância Magnética , Dados de Sequência MolecularRESUMO
We compared the disaccharide composition of dermatan sulfate (DS) purified from the ventral skin of three species of rays from the Brazilian seacoast, Dasyatis americana, Dasyatis gutatta, Aetobatus narinari and of Potamotrygon motoro, a fresh water species that habits the Amazon River. DS obtained from the four species were composed of non-sulfated, mono-sulfated disaccharides bearing esterified sulfate groups at positions C-4 or C-6 of N-acetyl galactosamine (GalNAc), and disulfated disaccharides bearing esterified sulfate groups at positions C-2 of the uronic acid and at position C-4 or C-6 of GalNAc. However, DS from the skin of P. motoro presented a very low content of the disulfated disaccharides. The anticoagulant actions of ray skin DS, measured by both APTT clotting and HCII-mediated inhibition of thrombin assays, were compared to that of mammalian DS. DS from D. americana had both high APTT and HCII activities, whereas DS from D. gutatta showed activity profiles similar to those of mammalian DS. In contrast, DS from both A. narinari and P. motoro had no measurable activity in the APTT assay. Thus, the anticoagulant activity of ray skin DS is not merely a consequence of their charge density. We speculate that the differences among the anticoagulant activities of these three DS may be related to both different composition and arrangements of the disulfated disaccharide units within their polysaccharide chains.