Effects of phospholipids on sphingomyelin hydrolysis induced by intestinal alkaline sphingomyelinase: An in vitro study Jian-Jun Liu, Åke Nilsson, and Rui-Dong Duan Department of Cell Biology 1, Department of Medicine, University Hospital of Lund, Lund, Sweden Digestion of dietary sphingomyelin (SM) is catalyzed by intestinal alkaline sphingomyelinase (SMase) and may have important implications in colonic tumorigenesis. Previous studies demonstrated that the digestion and absorption of dietary SM was slow and incomplete and that the colon was exposed to SM and its hydrolytic products including ceramide. In the present work, we studied the influences of glycerophospholipids and hydrolytic products of phosphatidylcholine (PC; i.e., lyso-PC, fatty acid, diacylglycerol, and phosphorylcholine) on SM hydrolysis induced by purified rat intestinal alkaline SMase in the presence of 10 mM taurocholate. It was found that various phospholipids including PC, phosphatidylserine (PS), phosphatidylinositol (PI), phosphati- dylethanolamine (PE), and phosphatidic acid (PA) inhibit alkaline SMase activity in a dose-dependent manner, with the degree of inhibition being in the order PA PS PI PC PE. Similar inhibition was also seen in a buffer of pH 7.4, which is close to the physiologic pH in the middle of the small intestine. When the effects of hydrolytic products of PC were studied, lyso-PC, oleic acid, and 1,2-dioleoyl glycerol also inhibited alkaline SMase activity, whereas phosphorylcholine enhanced SMase activity. However, in the absence of bile salt, acid phospholipids including PA, PS, and PI mildly stimulated alkaline SMase activity whereas PC and PE had no effect. It is concluded that in the presence of bile salts, glycerophospholipids and their hydrolytic products inhibit intestinal alkaline SMase activity. This may contribute to the slow rate of SM digestion in the upper small intestine. (J. Nutr. Biochem. 11:192–197, 2000) © Elsevier Science Inc. 2000. All rights reserved. Keywords: sphingomyelin digestion; alkaline sphingomyelinase; phosphatidylcholine; phosphatidylinositol; phosphatidylethanolamine; phosphatidylserine; phosphatidic acid; diacylglycerol; fatty acid; phosphorylcholine Introduction Sphingomyelin (SM) is not only a constituent of cell membranes but also a dietary component. In milk and most dairy products, SM is the major phospholipid, the amount being equal to or higher than that of phosphatidylcholine (PC). 1–3 SM is also present to a considerable extent in eggs, meat, and fish. 4 Recent studies have indicated that digestion of SM may have important implications in colonic tumori- genesis. 5 Administration of SM has been shown to reduce the number of aberrant colonic crypt foci and to decrease the proportion of carcinomas to adenomas induced by 1,2-dimethylhydrazine. 6,7 A significant decrease in sphin- gomyelinase (SMase) activity has been recently demon- strated to be associated with human colonic adenoma, carcinoma, and adenomatous polyposis. 8,9 SM is hydrolyzed by SMase and at least three types of SMase have been identified based on their optimal pH values. Acid SMase is a lysosomal enzyme with an optimal pH of 4.5, and neutral SMase is a membrane-bound enzyme with an optimal pH of 7.5. 10 Both acid and neutral SMases are common intracellular enzymes and it is unlikely that their role in SM digestion in the intestinal lumen are important. Nilsson 11 identified in both rat intestinal tract and human intestinal content a unique SMase, which was a brush-border enzyme and preferred an alkaline pH of 9.0. Address correspondence to Dr. Rui-Dong Duan, Department of Cell Biology 1, EB-Blocket, University Hospital of Lund, S-221 85 Lund, Sweden Received September 22, 1999; accepted January 5, 2000. J. Nutr. Biochem. 11:192–197, 2000 © Elsevier Science Inc. 2000. All rights reserved. 0955-2863/00/$–see front matter 655 Avenue of the Americas, New York, NY 10010 PII S0955-2863(00)00064-4