Résumé:
Although the field of biotechnology has developed extraordinarily in recent years, therapies
remain ineffective and can cause side effects in patients, which is why the laboratories have turned to the search for new natural products. Helix aspersa is a terrestrial mollusk that is widespread in Mediterranean countries and is part of the food habitude. It is an important source of protein and mineral salts and has several therapeutic effects known for centuries.
One of the main prospects of our study is the evaluation of the anti-inflammatory, anti-tumor and anti-angiogenic effects. Three extracts were prepared using solvents with different
polarity: an aqueous extract, an hydroalcoholic extract and an organic extract. Then, a model
of acute pulmonary inflammation was developed to evaluate in vivo the anti-inflammatory
activity of extracts by measuring total white blood cells and white blood cells in
bronchoalveolar fluid, myeloperoxydase activity and analyzing the histology of pulmonary
tissue. Analysis of the results showed that the extracts had anti-inflammatory activity by
decreasing the total leukocyte count in blood and in bronchoalveolar fluid, protecting the
pulmonary tissue by modulating neutrophil activity and decreasing protein concentration in
bronchoalveolar fluid. The antiproliferative activity of the extracts was evaluated by trypan
blue and the MTT test on several cell lines: MK2, A549, HUH7, K562, HUT-78 and SeAx.
The chromatin condensation and nuclear fragmentation were evaluated by the Hoechst test in
the HUT-78 and SeAx tumor cell line stimulated by the aqueous extract. The aqueous and
hydroalcoholic extracts had antiproliferative activity against the HUT-78 and SeAx lines. The
aqueous extract induced apoptosis in HUT-78 cells. The hydroalcoholic and organic extracts
had an antiproliferative activity on the HUH7 line. The three extracts were not toxic on the
MK2 normal cell line. The antiangiogenic effect was evaluated by measuring the expression
of MMP-9 in the two HUT-78 and SeAx lines stimulated in the presence of the aqueous and
hydroalcoholic extracts using was the zymography method. The aqueous and hydroalcoholic
extracts had no effect on the production of MMP-9. At the end, we extracted the soluble
proteins from meat lyophilized and we fractionated the extracts by anions exchange
chromatography. The proteolytic activity using synthetic substrates and protease inhibitors as
well as the antiproliferative activity of the different fractions obtained were measured. One of
the fractions contains proteases and has antiproliferative activity against the K562 liquid line.
Nevertheless, other contained no enzymes and had a toxic effect on the matrix tumor lines
A549 and HUH7. It was concluded that the aqueous, hydroalcoholic and organic extracts of
Helix aspersa had an anti-inflammatory and an anti-tumor effects, but did not have an antiangiogenic effect (on the expression of MMP-9)
