Análise estrutural e avaliação do efeito condroitim sulfato extraído de tilápia (Oreochromis niloticus) em modelo de peritonite aguda
Chondroitin sulfate (CS) is a naturally glycosaminoglycan found in the extracellular matrix of connective tissues and it may be extracted and purified those tissues. CS is involved in various biological functions, which may be related to the having structural variability, despite the simplicity o...
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| Formato: | Dissertação |
| Idioma: | por |
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Universidade Federal do Rio Grande do Norte
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| Endereço do item: | https://repositorio.ufrn.br/jspui/handle/123456789/19791 |
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| Resumo: | Chondroitin sulfate (CS) is a naturally glycosaminoglycan found in the extracellular
matrix of connective tissues and it may be extracted and purified those tissues. CS is
involved in various biological functions, which may be related to the having structural
variability, despite the simplicity of the linear chain structure from this molecule.
Researches in biotechnology and pharmaceutical field with wastes from aquaculture
has been developed in Brazil. In recent decades, tilapia (Oreochromis niloticus),
native fish from Africa, has been one of the most cultivated species in various regions
of the world, including Brazil. The tilapia farming is a cost-effective activity, however,
it generates large amount of wastes that are discarded by producers. It is understood
that waste from tilapia can be used in research as a source of molecules with
important biotechnological applications, which also helps in reducing environmental
impacts and promote the development of an ecofriendly activity. Thus, nile tilapia
viscera were subjected to proteolysis, then the glycosaminoglycans were complexed
with ion exchange resin (Lewatit), it was fractionated with increasing volumes of
acetone and purified by ion exchange chromatography DEAE-Sephacel. Further, the
fraction was analyzed by agarose gel electrophoresis and nuclear magnetic
resonance (NMR). The electrophoretic profile of the compound together the analysis
of 1H NMR spectra and the HSQC correlation allow to affirm that the compound
corresponds to a molecule like chondroitin sulfate. MTT assay was used to assess
cell viability in the presence of CS tilapia isolated and showed that the compound is
not cytotoxic to normal cells such as cells from the mouse embryo fibroblast (3T3).
Then, this compound was tested for the ability to reduce the influx of leukocytes in
model of acute peritonitis (in vivo) induced by sodium thioglycolate. In this context, it
was done total and differential leukocytes counting in the blood and peritoneal fluid
collected respectively from vena cava and the peritoneal cavity of the animals
subjected to the experiment. The chondroitin sulfate for the first time isolated from
tilapia (CST
) was able to reduce the migration of leukocytes to the peritoneal cavity of
inflamed mice until 80.4 per cent at a dose 10µg/kg. The results also show that there
was a significant reduction (p<0.001) of the population of polymorphonuclear
leukocytes from peritoneal cavity in the three tested doses (0.1µg/kg; 1µg/kg and
10µg/kg) when it was compared to the positive control (just thioglycolate). Therefore,
since the CST structure and mechanism of action has been completely elucidated,
this compound may have potential for therapeutic use in inflammatory diseases |
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