Phosphorylated glucosamine inhibits adipogenesis in 3T3-L1 adipocytes.
Phosphorylated glucosamine (glucosamine-6-phosphate, PGlc) was synthesized using methanesulfonic acid, phosphorus pentoxide (P(2)O(5)), NH(2)NH(2) and DMF. Its inhibitory effect on lipid accumulation in cultured 3T3-L1 adipocytes was investigated by measuring triglyceride contents and Oil Red O staining. In order to understand the mechanism by which lipid accumulation in adipocytes is decreased by PGlc, we examined the expression levels of several genes and proteins associated with adipogenesis and lipolysis using reverse transcription polymerase chain reaction, real-time polymerase chain reaction and Western blot analysis. Treatment with PGlc significantly reduced lipid accumulation during adipocyte differentiation and induced down-regulation of peroxisome proliferator-activated receptor-gamma, sterol regulatory element binding protein 1 and CCAAT/enhancer binding protein-alpha in a dose-dependent manner. Moreover, treatment with PGlc during adipocyte differentiation induced significant up-regulation of preadipocyte factor 1 mRNA and down-regulation of such adipocyte-specific gene promoters as adipocyte fatty acid binding protein, fatty acid synthase, lipoprotein lipase and leptin. According to the lipolytic response, PGlc up-regulated hormone-sensitive lipase mRNA expression and suppressed the expression levels of tumor necrosis factor-alpha mRNA compared with fully differentiated adipose tissue. These results suggest that the inhibitory effect of PGlc on adipocyte differentiation might be mediated through the down-regulation of adipogenic transcription factors, such as peroxisome proliferator-activated receptor-gamma, sterol regulatory element binding protein 1 and CCAAT/enhancer binding protein-alpha, which are related to the downstream adipocyte-specific gene promoters.
Kong
Chang-Suk
CS
Marine Bioprocess Research Center, Pukyong National University, Busan, South Korea.
Kim
Jung-Ae
JA
Eom
Tae Kil
TK
Kim
Se-Kwon
SK
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D050156:Adipogenesis / Q000187:drug effects*; D000818:Animals; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics / Q000378:metabolism; D002135:Calcium-Binding Proteins; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D019298:Glucose-6-Phosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D050356:Lipid Metabolism / Q000187:drug effects; D008066:Lipolysis / Q000187:drug effects; D051379:Mice; D009928:Organ Specificity; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D002787:Sterol Esterase / Q000235:genetics / Q000378:metabolism; D051780:Sterol Regulatory Element Binding Protein 1 / Q000235:genetics / Q000378:metabolism; D014280:Triglycerides / Q000032:analysis; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D022763:CCAAT-Enhancer-Binding Protein-alpha; D002135:Calcium-Binding Proteins; C512925:Dlk1 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D047495:PPAR gamma; D012333:RNA, Messenger; C497388:Srebf1 protein, mouse; D051780:Sterol Regulatory Element Binding Protein 1; D014280:Triglycerides; D014409:Tumor Necrosis Factor-alpha; C001293:glucosamine 6-phosphate; D019298:Glucose-6-Phosphate; D002787:Sterol Esterase; D005944:Glucosamine