Nandini D. Chilkunda

Our earlier study demonstrated that metabolic disorders increase the expression of Syndecan‐3 (Sdc‐3), a heparan sulfate proteoglycan (HSPG) in erythrocytes, contributing to adhesion through the glycosaminoglycan chain. Reactive oxygen species (ROS) could be one of the factors for increased expression. This study aimed to determine whether quercetin, a bioactive antioxidant commonly found in food, could modulate Sdc‐3 expression. Male Wistar rats were made dyslipidemic and diabetic, after which they were treated with quercetin at doses of 50 and 100 mg/kg body weight for a duration of 2 months. Sdc‐3 expression in erythrocytes was assessed by Western blot, and erythrocyte adhesion to fibronectin was evaluated in‐vitro. The quercetin‐supplemented diet reduced circulating lipids, blood glucose, and MDA levels, mitigated changes in Sdc‐3 expression, and decreased erythrocyte adhesion to fibronectin. These effects may be attributed to quercetin′s antioxidant properties, as was seen by the positive correlation between MDA levels and Sdc‐3 expression. Sdc‐3 levels in erythrocytes could serve as a prognostic marker for assessing the impact of dietary compounds on complications linked to metabolic disorders.

Glycosaminoglycans (GAGs) are essential bone extracellular matrix molecules that regulate osteoblast differentiation. Numerous studies have explored endogenous and exogenous GAG osteoanabolic activities using appropriate in vitro and in vivo models. However, GAGs’ underlying the mechanism of action and structure-function relationships need to be elucidated in detail. Earlier, we showed that exogenous GAG can bring about osteogenesis in pre-osteoblast cells. In the present study, we have elucidated the mechanism of action of exogenous GAGs, especially of the chondroitin sulfate/dermatan sulfate (CS/DS) class on osteogenesis. GAGs were immobilized, and osteoblast differentiation was evaluated in MC3T3-E1 cells. Results indicated that GAGs supported osteoblast differentiation by promoting collagen production, extracellular matrix formation, and subsequent mineralization. We elucidated the mechanisms underlying these effects by assessing the key signaling molecules involved in osteogenesis in response to exogenous CS/DS with/without BMP2. CS/DS alone significantly increased pERK1/2 and ATF4 expression levels differentially in a time-dependent manner without significant effects on BMP2, RUNX2, and pSMAD5 protein expression. On the other hand, CS/DS, in the presence of BMP2, differentially increased BMP2, pSMAD5, pERK1/2, RUNX2, and ATF4 expression levels at various time points. Collectively, these results strongly suggest that CS/DS can promote osteogenesis, and in the presence of BMP2, it could promote SMAD-mediated ERK-dependent osteogenesis.

O-GlcNAcylation, a nutritionally driven, post-translational modification of proteins, is gaining importance because of its health implications. Changes in O-GlcNAcylation are observed in various disease conditions. Changes in O-GlcNAcylation by diet that causes hypercholesterolemia are not critically looked into in the liver. To address it, both in vitro and in vivo approaches were employed. Hypercholesterolemia was induced individually by feeding cholesterol (H)/high-fat (HF) diet. Global O-GlcNAcylation levels and modulation of AMPK activation in both preventive and curative approaches were looked into. Diet-induced hypercholesterolemia resulted in decreased O-GlcNAcylation of liver proteins which was associated with decreased O-linked N-acetylglucosaminyltransferase (OGT) and Glutamine fructose-6-phosphate amidotransferase-1 (GFAT1). Activation of AMPK by metformin in preventive mode restored the O-GlcNAcylation levels; however, metformin treatment of HepG2 cells in curative mode restored O-GlcNAcylation levels in HF but failed to in H condition (at 24 h). Further, maternal faulty diet resulted in decreased O-GlcNAcylation in pup liver despite feeding normal diet till adulthood. A faulty diet modulates global O-GlcNAcylation of liver proteins which is accompanied by decreased AMPK activation which could exacerbate metabolic syndromes through fat accumulation in the liver. Graphical abstract

In utero hyperglycemia has consequences on future outcomes in the offsprings. We had earlier shown that in utero hyperglycemia impacts proteoglycans/glycosaminoglycans, one of the key molecules involved in brain development. Hypothalamic HSPGs such as syndecan-1 and syndecan-3 are well known for their involvement in feeding behavior. Therefore, studies were carried out to determine the effect of maternal hyperglycemia on the expression of HSPGs in the hypothalamus of offspring brain. Results revealed increased protein abundance of Syndecan-1 and -3 as well as glypican-1 in postnatal adults from hyperglycemic mothers. This was associated with increased hyperphagia and increased expression of Neuropeptide Y. These results indicate the likely consequences on offsprings exposed to in utero hyperglycemia on its growth. Graphical abstract

Glycosaminoglycans (GAGs) and AMP-activated protein kinase (AMPK) are two critical molecular players involved in cellular homeostasis. Both of them are altered due to hyperglycaemia in the kidney, leading to the pathogenesis of diabetic nephropathy. Here, we have looked into the effect of AMPK modulation on sulphated GAG (sGAG) levels of tubular cells of proximal and distal origin to understand the mechanism of hyperglycaemia-mediated pathogenesis of the diabetic nephropathy. In MDCK cells (distal tubular cell) and NRK-52E (proximal tubular cell), AMPK inhibition resulted in increased sGAG levels under normal glucose conditions characteristically of heparan sulphate class, whereas AMPK activation did not have any effect. High glucose (HG) condition did not alter sGAG levels in MDCK cell despite a decrease in AMPK phosphorylation. Subjecting NRK-52E cells to HG milieu significantly decreased sGAG levels more so of chondroitin/dermatan sulphate, which is significantly prevented when HG is co-treated with AMPK activator. Interestingly, knockdown of AMPK by AMPKα1/α2 siRNA showed increased sGAG levels in NRK-52E. Our results suggest that changes in sGAG level, in particular, as a result of AMPK modulation is differentially regulated and is dependent on cell type as well as its physiological status. Furthermore, activation of AMPK is beneficial in preventing the HG-mediated decrease in sGAGs in proximal tubular cells.