Occurrence of sulfated fucose branches in fucosylated chondroitin sulfate are essential for the polysaccharide effect preventing muscle damage induced by toxins and crude venom from Bothrops jararacussu snake

Abstract

Snake envenoming is an important public health problem around the world, particularly in tropics. Beyond deaths, morbidity induced by snake venoms, such as myotoxicity, is of pivotal consequence to population. Bothrops jararacussu is the main venomous snake in southeast region of Brazil, and particularly presents strong myotoxic effect. The only available therapy, antibothropic antivenom, poorly affects venom-induced myotoxicity. The aim of this study is to assess the ability of fucosylated chondroitin sulfate (fucCS), a glycosaminoglycan with anticoagulant and antithrombotic properties, and its derivatives to inhibit toxic activities of B. jararacussu crude venom and its isolated toxins, named bothropstoxins (BthTX-I and BthTX-II). The in vitro myotoxic activities induced by crude venom, by BthTX-I alone and by toxins together were abolished by fucCS. Carboxyl reduction (fucCS-CR) kept this ability whereas defucosilation (defucCS) abrogates myoprotection. We observed the same pattern in the response of these polysaccharides in antagonizing the increase in plasma creatine kinase (CK) levels, the reduction of skeletal muscle CK content and the rise of myeloperoxidase (MPO) activity induced by crude venom and isolated toxins. FucCS inhibited edematogenic activity and partially prevented the reduction of total leukocytes in blood when pre-incubated with crude venom. Furthermore, the venom procoagulant effect was completely antagonized by increasing concentrations of fucCS, although this polyanion could stop neither the tail bleeding nor the skin hemorrhage induced by Bothrops jararaca venom. The B. jararacussu phospholipase, hyaluronidase, proteolytic and collagenase activities were inhibited in vitro. The results suggest that fucCS could be able to interact with both toxins, and it is able to inhibit BthTX-II phospholipase activity. Light microscopy of extensor digitorum longus muscle (EDL) muscle showed myoprotection by fucCS, once necrotic areas, edema and inflammatory cells were all decreased as compared to venom injection alone. Altogether, data show that fucCS was able to inhibit myotoxicity and inflammation induced by B. jararacussu venom and its phospholipase toxins, BthTX-I and BthTX-II. Thus, fucosylated chondroitin sulfate is a new polyanion with potential to be used as an adjuvant in the treatment of snakebites in the future.

Involvement of Nitric Oxide on Bothropoides insularis Venom Biological Effects on Murine Macrophages In Vitro

“…Moreover, it is known that inflammation plays an important role in the development of snake venom toxicity. It is closely associated to the onset of local and systemic toxicity induced by bothropic venom and its fractions, as described by several authors [ 11 – 13 ]. Additionally, some proteins in these venoms have shown effects on cells associated to inflammation, such as macrophages [ 14 – 17 ] and neutrophils [ 15 , 18 ].…”

A Study for the Access to a Semi-synthetic Regioisomer of Natural Fucosylated Chondroitin Sulfate with Fucosyl Branches on N-acetyl-Galactosamine Units

“…These features make fCS a very promising anticoagulant drug candidate for heparin replacement [6].From a structural point of view, fCS shares the same linear core backbone as chondroitin sulfate (CS) polysaccharide, with alternating N-acetyl-d-galactosamine (GalNAc) and d-glucuronic acid (GlcA) residues linked together through alternating β-1→3 and β-1→4 glycosidic bonds and sulfated to a different extent on their hydroxyls. The unique structural peculiarity of fCSs is the additional presence of variously sulfated fucose (Fuc) branches [7,8], which are essential for the observed biological activities [9][10][11]. Very often the branches are constituted of a single Fuc unit α-glycosidically linked at O-3 site of GlcA residues.…”

Sulfated Galactan from Palisada flagellifera Inhibits Toxic Effects of Lachesis muta Snake Venom

“…Crude extracts of plants or their isolated molecules with antivenom property have been reported [ 33 , 34 ], but little investigation has been performed with seaweeds [ 35 ] and, even less with the polysaccharides from such source [ 24 , 25 ]. Anticoagulant and antiviral effects of sulfated polysaccharides from seaweeds are well established and, in some cases a possible correlation between chemical structure and mechanism of action has been proposed [ 36 , 37 , 38 , 39 , 40 ]. Furthermore, some reports describe the neutralizing capacity displayed by sulfated fucoidans from brown seaweeds against snake venoms [ 25 ].…”