Hydrolytic activities of human pancreatic phospholipase A2 and endotoxin-stimulated endogenous phospholipase A2 toward membrane phospholipids of erythrocytes.
The problem of whether human pancreatic phospholipase A2 (PLA2) can really hydrolyze membrane phospholipids in vitro was studied to understand pathophysiology of acute pancreatitis. Total amount of lysophospholipids generated in erythrocytes by exogenously added human pancreatic PLA2 (2 micrograms/ml) was only 12% of the amount of sphingomyelin, which was not decomposed by the enzyme. About fivefold the amount of lysophospholipids was generated in ghost membranes during one-sixth of the incubation time compared to that in intact erythrocyte membranes. Escherichia coli lipopolysaccharide (LPS) (10 micrograms/ml) was able to stimulate membrane-associated PLA2 of erythrocytes, the amount of lysophospholipids generated being 12.5% of that of sphingomyelin without adding the exogenous PLA2. The stimulation of membrane-associated PLA2 in erythrocytes was inhibited by pretreatment of lipopolysaccharide with polymyxin-B sulfate. When intact erythrocytes were incubated with human pancreatic PLA2 and LPS, the amount of generated lysophospholipids was 24% of that of sphingomyelin. These results suggested that the exogenously added human pancreatic PLA2 cannot degrade phospholipids of intact erythrocytes so extensively under physiological conditions, and, in acute pancreatitis, unknown factors may be involved in the hydrolysis of phospholipids. LPS, which activates membrane-associated PLA2, may be one of the factors, and thus membrane phospholipids are hydrolyzed in the disease.
D004731:Endotoxins / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism; D004926:Escherichia coli; D006801:Humans; D006868:Hydrolysis; D066298:In Vitro Techniques; D008070:Lipopolysaccharides / Q000494:pharmacology; D008246:Lysophospholipids; D008563:Membrane Lipids / Q000097:blood*; D010179:Pancreas / Q000378:metabolism*; D010195:Pancreatitis / Q000201:enzymology; D010740:Phospholipases / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D010743:Phospholipids / Q000097:blood*; D011112:Polymyxin B / Q000494:pharmacology
D004731:Endotoxins; D008070:Lipopolysaccharides; D008246:Lysophospholipids; D008563:Membrane Lipids; D010743:Phospholipids; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D011112:Polymyxin B