Lonomia obliqua (Lepidoptera, Saturniidae) caterpillar bristle extract induces direct lysis by cleaving erythrocyte membrane glycoproteins.
Lonomia obliqua caterpillar bristle extract induces hemolysis in vitro on washed human and rat erythrocytes, in either the absence or presence of exogenous lecithin. In the former condition, phospholipases A(2) are key enzymes involved in hemolysis. However, the mechanism whereby this extract causes direct hemolysis is not known. Thus, the aim of this study was to investigate the hemolytic mechanism of the crude extract of the caterpillar L. obliqua on human erythrocytes in the absence of lecithin. The extract significantly increased the erythrocyte osmotic fragility and promoted the removal of glycophorins A and C, and band 3 from the erythrocyte membrane. The use of Ca2+ and Mg2+ ions significantly potentiated glycoprotein removal, remarkably of erythrocyte band 3. The composition of fatty acids was analyzed by HPLC in both L. obliqua caterpillar bristle extract and human erythrocyte membranes incubated with the extract. The levels of unsaturated fatty acids were remarkably augmented in erythrocytes incubated with the extract than in control erythrocytes, modifying thereby the saturated/unsaturated fatty acid ratio. Altogether, evidence is provided here that the interplay of at least three mechanisms of action accounts for the direct activity of the bristle extract on erythrocyte membrane, leading to hemolysis: the removal of glycoproteins and band 3; the insertion of fatty acids; and the action of phospholipases. Such mechanisms might affect erythrocyte flexibility and deformability, which may induce hemolysis by increasing erythrocyte fragility. However, whether the direct hemolytic activity of L. obliqua caterpillar is the major cause of intravascular hemolysis during envenomation still needs further investigation.
Seibert
Carla Simone
CS
Núcleo de Estudos Ambientais, Universidade Federal do Tocantins, Campus de Porto Nacional, TO, Brazil.
Santoro
Marcelo L
ML
Tambourgi
Denise Vilarinho
DV
Sampaio
Sandra Coccuzzo
SC
Takahashi
Hilton Kenji
HK
Peres
Carmem Maldonado
CM
Curi
Rui
R
Sano-Martins
Ida S
IS
D000818:Animals; D000906:Antibodies / Q000737:chemistry; D002784:Cholesterol / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004910:Erythrocyte Membrane / Q000737:chemistry*; D005231:Fatty Acids, Unsaturated / Q000097:blood; D005434:Flow Cytometry; D006021:Glycophorins / Q000737:chemistry; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D007120:Immunochemistry; D066298:In Vitro Techniques; D007814:Larva / Q000737:chemistry*; D007915:Lepidoptera / Q000737:chemistry*; D008055:Lipids / Q000097:blood; D008562:Membrane Glycoproteins / Q000737:chemistry*; D009996:Osmotic Fragility / Q000187:drug effects; D010718:Phosphatidylserines / Q000633:toxicity; D054467:Phospholipases A2 / Q000737:chemistry / Q000633:toxicity; D051381:Rats; D014020:Tissue Extracts / Q000633:toxicity*; D014280:Triglycerides / Q000097:blood
D000906:Antibodies; D005231:Fatty Acids, Unsaturated; D006021:Glycophorins; D008055:Lipids; D008562:Membrane Glycoproteins; D010718:Phosphatidylserines; D014020:Tissue Extracts; D014280:Triglycerides; D002784:Cholesterol; D054467:Phospholipases A2