Qualitative and quantitative analyses in sulfated glycosaminoglycans, chondroitin sulfate/dermatan sulfate, during 3T3‐L1 adipocytes differentiation

Abstract

AbstractChondroitin sulfate/dermatan sulfate (CS/DS) is a member of glycosaminoglycans (GAGs) found in animal tissues. Major CS/DS subclasses, O, A, C, D, and E units, exist based on the sulfation pattern in d‐glucuronic acid (GlcA) and N‐acetyl‐d‐galactosamine repeating units. DS is formed when GlcA is epimerized into l‐iduronic acid. Our study aimed to analyze the CS/DS profile in 3T3‐L1 cells before and after adipogenic induction. CS/DS contents, molecular weight (Mw), and sulfation pattern were analyzed by using high‐performance liquid chromatography. CS/DS synthesis‐ and sulfotransferase‐related genes were analyzed by reverse transcription real‐time PCR. CS/DS amount was significantly decreased in the differentiated (DI) group compared to the non‐differentiated (ND) group, along with a lower expression of CS biosynthesis‐related genes, chondroitin sulfate N‐acetylgalactosaminyltransferase 1 and 2, as well as chondroitin polymerizing factor. GAGs in the DI group also showed lower Mw than those of ND. Furthermore, the A unit was the major CS/DS in both groups, with a proportionally higher CS‐A in the DI group. This was consistent with the expression of carbohydrate sulfotransferase 12 that encodes chondroitin 4‐O‐sulfotransferase, for CS‐A formation. These qualitative and quantitative changes in CS/DS and CS/DS‐synthases before and after adipocyte differentiation reveal valuable insights into adipocyte development.