High Glucose Alters Proteoglycan Expression and the Glycosaminoglycan Composition in Placentas of Women with Gestational Diabetes Mellitus and in Cultured Trophoblasts

Abstract 

Impaired glucose metabolism with diabetes may alter the expressions of proteoglycans (PGs), which may impair the biological functions of placenta. In this study, we investigated the expression of PGs and their conjugated glycosaminoglycan (GAG) composition in the placentas of mothers with gestational diabetes mellitus (GDM) and trophoblasts cultured in a high-glucose condition. The PGs by guanidine/HCl extraction and DEAE Sepharose fractionation followed by GAG degradation enzyme digestion analyses showed that the expression of chondroitin sulfate and/or dermatan sulfate (CS/DS) PGs was increased whereas the heparan sulfate (HS) PG was decreased in GDM placentas compared to controls. Western blot analyses demonstrated that the increased CS/DS PGs in GDM placentas were predominantly the small leucine-rich proteoglycans (SLRPs), decorin and biglycan. Increased mRNA expression level was consistently shown by quantitative real-time PCR. Immunohistochemistry indicated intensive staining of decorin and biglycan in the diabetic placenta with different localizations. Additionally, the basement membrane HSPG, perlecan was found to contain both CS/DS and HS in GDM placentas and plain HS in controls. Similar findings of PG alterations induced by hyperglycemia were observed in cultured trophoblast in a high-glucose condition. This study demonstrated that hyperglycemia induced not only the gene expressions of PGs but also alterations in the carried GAG type and composition.

Keywords: Biglycan, Decorin, Gestational diabetes mellitus, Glycosaminoglycan, Perlecan, Proteoglycan

Abbreviations: PG, proteoglycan, ECM, extracellular matrix, GDM, gestational diabetes mellitus, CS, chondroitin sulfate, DS, dermatan sulfate, SLRP, small leucine-rich proteoglycan, HS, heparan sulfate, GAG, glycosaminoglycan, KS, keratan sulfate, Hepase, heparintinase, Chase ABC, chondroitinase ABC, Chase B, chondroitinase B, CHAPS, 3-[(3)-cholamidopropyl-dimethyl-ammonio]-1-propanesulfonate, PMSF, phenylmethylsulfonyl fluoride, PBS, phosphate-buffered saline, CPC, 1-hexadecyl pyridinium chloride, SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis