title: Lysophosphatidylcholine-stimulated protein and glycoprotein production by human gallbladder mucosal cells.
abstract: It has been demonstrated in experimental cholecystitis in cats produced by lysophosphatidylcholine that the development of inflammation is associated with the exsorption of a large amount of protein into the gallbladder lumen. It was subsequently demonstrated that in feline experimental cholecystitis the protein produced was albumin and that its production was decreased by vesicular transport inhibitors, suggesting an active secretory process. In the present study, the effect of lysophosphatidylcholine on protein production by fresh, isolated human gallbladder mucosal cells was evaluated. Isolated gallbladder mucosal cells were incubated with [14C]leucine for 24 hr in tissue culture medium. The cells readily incorporated the radioactive label into cellular protein, a process inhibited by cycloheximide. Exposure of the cells to lysophosphatidylcholine for 1 hr in buffer solution resulted in loss of intracellular protein into the buffer solution. Exposure of the cells for 1 hr prior to lysophosphatidylcholine administration to vesicular transport inhibitors, colchicine, and cytochalasin B and to 4 degrees C culture conditions failed to alter the lysophosphatidylcholine-produced passage of the 14C label extracellularly. SDS-PAGE evaluation of the protein produced demonstrated that human gallbladder mucosal cells continuously produced a 66-kDa protein that was not increased by increasing concentration of lysophosphatidylcholine and a 14-kDa protein that increased with increasing concentrations of lysophosphatidylcholine. Employing Western blotting with specific antibodies, the 66-kDa protein was demonstrated to not be albumin but a 66-kDa glycoprotein, and the 14-kDa protein was demonstrated to contain phospholipase A2. Human gallbladder mucosal cells produced a protein and glycoprotein in response to lysophosphatidylcholine by a mechanism not related to vesicular transport.
authors.affiliation: Department of Surgery, St. Louis University Health Sciences Center, Missouri 63110-0250, USA.
authors.forename: M K
authors.initials: MK
authors.lastname: Nag
authors.forename: Y G
authors.initials: YG
authors.lastname: Deshpande
authors.forename: A
authors.initials: A
authors.lastname: Li
authors.forename: D
authors.initials: D
authors.lastname: Beck
authors.forename: D L
authors.initials: DL
authors.lastname: Kaminski
mesh_terms: D002250:Carbon Radioisotopes; D002478:Cells, Cultured; D005260:Female; D005704:Gallbladder / Q000166:cytology / Q000378:metabolism*; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007930:Leucine; D008244:Lysophosphatidylcholines / Q000494:pharmacology*; D008297:Male; D008875:Middle Aged; D009092:Mucous Membrane / Q000166:cytology / Q000378:metabolism; D010741:Phospholipases A / Q000032:analysis / Q000378:metabolism*; D054467:Phospholipases A2; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002250:Carbon Radioisotopes; D006023:Glycoproteins; D008244:Lysophosphatidylcholines; D011506:Proteins; D010741:Phospholipases A; D054467:Phospholipases A2; D007930:Leucine