Preliminary molecular characterization of a proinflammatory and nociceptive molecule from the Echinometra lucunter spines extracts

 

Juliana Mozer Sciani¹, Bianca Zychar², Luis Roberto Gonçalves², Renata Giorgi², Thiago Nogueira², Daniel Carvalho Pimenta¹ 

 

1 Laboratory of Biochemistry and Biophysics, Butantan Institute

2 Laboratory of Pathophysiology, Butantan Institute

 

ABSTRACT

Background

Sea urchins are animals commonly found on the Brazilian shoreline, being Echinometra lucunter the most abundant species. Accidents caused by E. lucunter have been reported as one of the most frequent in Brazil, and are characterized by intense pain and inflammation, consequence of spine puncture in the skin. In order to characterize such toxic effects, we isolated one molecule that caused inflammatory and nociceptive effects.

Methods

E. lucunter specimens were collected without gender distinction. Spines were removed and molecules were extracted, fractionated by RP-HPLC and assayed for inflammatory and nociceptive activity, in a biological-driven fractionation way, until the obtainment of one active molecule and its subsequent analysis by mass spectrometry (MS and MS/MS). For inflammation, intravital microscopy was performed on the mouse cremaster muscle, in order to evaluate rolled, adherent and migrating leukocytes. Paw edema was also evaluated. For the nociceptive activity, the paw pressure test was performed in rats.

Results

One molecule could be isolated and related to the inflammatory and nociceptive activity. Regarding inflammation, increase in adherent and migrating cells was observed in the cremaster muscle after the administration of the molecule. Corroborating the inflammatory response, paw edema was also observed, although only in 20% of controls and 20 min after injection. Additionally, this molecule was able to decrease significantly the pain threshold, characterizing hyperalgesia. This molecule was analyzed by mass spectrometry, and according to the exact molecular mass, isotopic distribution and fragmentation profile, it was possible to propose the molecular formula C₂₉H₄₈N₃O₁₀.

Conclusions

One isolated molecule from the spine extract of E. lucunter is able to elicit inflammation and hypernociception in animal models, which is in agreement with the effects observed in sea urchin accidents.

 

Key words: Toxins; Sea urchin; Echinometra lucunter; Spines; Inflammation

 

Funding

The authors acknowledge the financial support of FINEP (Convênio FINEP — 01.09.0278.00). DCP is a fellow CNPq researcher (303792/2016–7).

 

Received: May 12, 2017.

Revised: September 15, 2017.

Accepted: October 3, 2017.

 

Correspondence: jmsciani@butantan.gov.br

 

Authors’ contributions

JMS and DCP performed the collection of animals, purification and analysis of molecules. BCZ and LRG designed and conducted intravital microcirculation and edema studies. RG and TON planned and conducted hyperalgesic assays. All authors contributed to the writing of the text, and read and approved the final manuscript.

 

Competing interests

The authors declare that there are no competing interests.

 

Ethics approval and consent to participate

Specimens of E. lucunter were collected under license number 13852–1 from the Brazilian Environmental Agency (IBAMA). All procedures involving mice and rats were in accordance with the guidelines for animal experimentation and were approved by the Institutional Animal Care Committee of the Butantan Institute (CEUAIB, protocol number 438/07).

 

Consent for publication

Not applicable

 

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.