Characterization of the de novo biosynthetic enzyme of platelet activating factor, DDT-insensitive cholinephosphotransferase, of human mesangial cells.
Platelet activating factor (PAF), a potent inflammatory mediator, is implicated in several proinflammatory/inflammatory diseases such as glomerulonephritis, glomerulosclerosis, atherosclerosis, cancer, allergy, and diabetes. PAF can be produced by several renal cells under appropriate stimuli and it is thought to be implicated in renal diseases. The aim of this study is the characterization of DTT-insensitive cholinephosphotransferase (PAF-CPT) of human mesangial cell (HMC), the main regulatory enzyme of PAF de novo biosynthetic pathway. Microsomal fractions of mesangial cells were isolated and enzymatic activity and kinetic parameters were determined by TLC and in vitro biological test in rabbit washed platelets. The effect of bovine serum albumin (BSA), dithiothreitol (DTT), divalent cations (Mg2+ and Ca2+), EDTA, and various chemicals on the activity of PAF-CPT of HMC was also studied. Moreover, preliminary in vitro tests have been performed with several anti-inflammatory factors such as drugs (simvastatin, IFNa, rupatadine, tinzaparin, and salicylic acid) and bioactive compounds of Mediterranean diet (resveratrol and lipids of olive oil, olive pomace, sea bass "Dicentrarchus labrax," and gilthead sea bream "Sparus aurata"). The results indicated that the above compounds can influence PAF-CPT activity of HMC.
Tsoupras
Alexandros Basilios
AB
Faculty of Chemistry, School of Sciences, National and Kapodistrian University of Athens, Panepistimioupolis, 15771 Athens, Greece.
Fragopoulou
Elizabeth
E
Nomikos
Tzortzis
T
Iatrou
Christos
C
Antonopoulou
Smaragdi
S
Demopoulos
Constantinos Alexandros
CA
D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D003634:DDT / Q000494:pharmacology*; D002798:Diacylglycerol Cholinephosphotransferase / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D050527:Mesangial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010972:Platelet Activating Factor / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D012969:Sodium Fluoride / Q000494:pharmacology; D013450:Sulfones / Q000494:pharmacology; D013696:Temperature
D010972:Platelet Activating Factor; D013450:Sulfones; D012710:Serum Albumin, Bovine; C002010:4-(2-aminoethyl)benzenesulfonylfluoride; D012969:Sodium Fluoride; D004492:Edetic Acid; D003634:DDT; D002798:Diacylglycerol Cholinephosphotransferase; D002118:Calcium; D004229:Dithiothreitol
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