Chondroitin 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 3 T3-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.

中文翻译：

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3 个 T3-L1 脂肪细胞分化过程中硫酸化糖胺聚糖、硫酸软骨素/硫酸皮肤素的定性和定量分析

硫酸软骨素/硫酸皮肤素 (CS/DS) 是动物组织中发现的糖胺聚糖 (GAG) 的成员。主要的 CS/DS 亚类、O、A、C、D 和 E 单元基于d-葡萄糖醛酸 (GlcA) 和N-乙酰基-d-半乳糖胺重复单元中的硫酸化模式而存在。当 GlcA 差向异构化为L-艾杜糖醛酸时，形成 DS 。我们的研究旨在分析成脂诱导前后 3 个 T3-L1 细胞的 CS/DS 谱。采用高效液相色谱法分析 CS/DS 含量、分子量 (Mw) 和硫酸化模式。通过逆转录实时 PCR 分析 CS/DS 合成和磺基转移酶相关基因。与非分化（ND）组相比，分化（DI）组的CS/DS量显着降低，并且CS生物合成相关基因、硫酸软骨素N-乙酰半乳糖氨基转移酶1和2以及软骨素的表达降低聚合因子。DI 组中的 GAG 的 Mw 也低于 ND 组中的 Mw。此外，A 单位是两组中主要的 CS/DS，DI 组中的 CS-A 比例较高。这与碳水化合物磺基转移酶 12 的表达一致，该酶编码软骨素 4- O-磺基转移酶，用于 CS-A 的形成。脂肪细胞分化前后 CS/DS 和 CS/DS 合酶的这些定性和定量变化揭示了对脂肪细胞发育的有价值的见解。