Localization Of Advanced Glycation End Products Research Articles

Localization of advanced glycation end-products and their receptor in tendinopathic lesions.

This study was designed to investigate the accumulation of advanced glycation end-products (AGEs) and the expression of the receptor of AGEs (RAGE) in tendinopathic tissues. In this study, tendinopathic posterior tibial tendons (PTT) were collected from patients (n=6). Redundant autografts of flexor digitorum longus tendon (FDL; n=3) were used for controls. The control and tendinopathic tendon tissues were used for extraction of proteins for western blot and sectioned for histology and immunohistochemistry. Tendinopathy of the PTT was confirmed histologically by the presentation of disorderly organized collagen fibers, high cellularity and increased vascularity. By immunohistochemistry, heterogeneous accumulation of AGEs was detected on the PTT sections and concentrated in areas, where collagen fibers were disorderly and tangled. In the PTT, roundish tenocytes were also AGEs-positive. In contrast, AGEs were diffuse, lightly stained in the FDL. A greater number of tenocytes within the tendinopathic lesions in the PTT were RAGE positive, compared to the tenocytes in the FDL. Western blot confirmed the expression of AGEs and RAGE in both tendinopathic PTT and control FDL but their band densities were not significantly different. The spatial relation of the accumulated AGEs and RAGE- positive tenocytes within the tendinopathic lesions indicates their involvement in the molecular pathology of tendinopathy.

The specific localization of advanced glycation end-products (AGEs) in rat pancreatic islets

Advanced glycation end-products (AGEs) are produced by non-enzymatic glycation between protein and reducing sugar such as glucose. Although glyceraldehyde-derived AGEs (Glycer-AGEs), one of the AGEs subspecies, have been reported to be involved in the pathogenesis of various age-relating diseases such as diabetes mellitus or arteriosclerosis, little is known about the pathological and physiological mechanism of AGEs in vivo. In present study, we produced 4 kinds of polyclonal antibodies against AGEs subspecies and investigated the localization of AGEs-modified proteins in rat peripheral tissues, making use of these antibodies. We found that Glycer-AGEs and methylglyoxal-derived AGEs (MGO-AGEs) were present in pancreatic islets of healthy rats, distinguished clearly into the pancreatic α and β cells, respectively. Although streptozotocin-induced diabetic rats suffered from remarkable impairment of pancreatic islets, the localization and deposit levels of the Glycer- and MGO-AGEs were not altered in the remaining α and β cells. Remarkably, the MGO-AGEs in pancreatic β cells were localized into the insulin-secretory granules. These results suggest that the cell-specific localization of AGEs-modified proteins are presence generally in healthy peripheral tissues, involved in physiological intracellular roles, such as a post-translational modulator contributing to the secretory and/or maturational functions of insulin.

Effects of exercise on the nephron of Goto-Kakizaki rats: morphological, and advanced glycation end-products and inducible nitric oxide synthase immunohistochemical analyses

The current study aimed to examine how exercise affects morphology of the nephron, and localization of advanced glycation end-products (AGEs) and inducible nitric oxide synthase (iNOS) immunoreactivity in diabetic Goto-Kakizaki rats. Four groups of male rats were studied. WIS SED (Wistar rats; sedentary) group served as a control. Other groups were WIS EX (Wistar rats; exercise), GK SED (Goto-Kakizaki diabetic rats; sedentary) and GK EX (Goto-Kakizaki diabetic rats; exercise) groups. The rats in EX groups were subjected to 15weeks of treadmill running at a speed of 15m/min for a total of 30minutes, three times a week. Changes in the structure of renal corpuscles and in the distribution of AGEs- and iNOS-immunoreactive cells of the uriniferous tubules were evaluated. Every parameter of GK EX was significantly different from that of GK SED (area of Bowman's capsules: p<0.001, area of glomeruli: p<0.05 and the occupancy of a glomerulus: p<0.05). These findings suggest that exercise may ameliorate glomerular filtration rate (GFR). The localizations of AGEs and iNOS immunostaining in the uriniferous tubules were similar in each group. Immunohistochemical assays revealed that the number of the AGEs and iNOS immunopositive cells of the proximal tubule of cortico-deep layer in EX groups were markedly greater than those in SED groups and that iNOS expression in GK EX was significantly higher than GK SED (p<0.05). Exercise seems to normalize the GFR and glomerular filtrate absorption from the uriniferous tubules in Goto-Kakizaki diabetic rats with the recovered shape of renal corpuscles and may be involved in the absorption and catabolization of AGEs with iNOS-related reactions for reabsorption.

Editorial Comment

This manuscript is the first study that has demonstrated the immunohistochemical localization of advanced glycation endproducts (AGE) in the human bladder. The conclusion is that AGE-structures such as Nε-(carboxymethyl)lysine (CML) and pentosidine were accumulated extracellularly in the human bladder, and then they were endocytosed by tissue macrophages. The successive researcher may cite this report as the first paper that has demonstrated the localization of AGE in the human bladder. The figures (especially1a) are not accepted without some criticism. Although the authors mention that a significant deposition of CML was observed in the connective tissue between the muscles, the localization of CML is not clearly recognizable in figure 1a. In histocytochemical studies, the figures are most important. Furthermore, a strict control study should be performed, especially in the first study.1 In conclusion, further investigation by the authors is widely expected.

Immunohistochemical localization of advanced glycation end-products (AGEs) and their receptor (RAGE) in polycystic and normal ovaries

The aim of the present study was to investigate the localization/immunohistochemical distribution of AGEs and RAGE, as well as their putative signalling mediator NF-kappaB in ovaries of women with polycystic ovary syndrome (PCOS) compared to normal. Archival ovarian-tissue samples from biopsies of six women with PCOS and from six healthy of similar age women, were examined immunohistochemically with monoclonal anti-AGEs, anti-RAGE and anti-NF-kappaB(p50/p65) specific antibodies. In healthy women, AGE immunoreactivity was observed in follicular cell layers (granulosa and theca) and luteinized cells, but not in endothelial cells. PCOS specimens displayed AGE immunoexpression in theca interna and granulosa cells as well as in endothelial cells, but staining of granulosa cells was stronger than in that of normal ovaries. RAGE was highly expressed in normal and PCOS tissues. Normal tissue exhibited no staining differences between granulosa cell layer and theca interna. However, in PCOS ovaries, granulosa cells displayed stronger RAGE expression compared to theca interna cells in comparison to controls. NF-kappaB(p50/p65) was expressed in the cytoplasm of theca interna and granulosa cells of both normal and PCOS ovaries; whereas the NF-kappaB p65 subunit was only observed in granulosa cells nuclei in PCOS tissue. In conclusion, these findings demonstrate for the first time that RAGE and AGE-modified proteins with activated NF-kappaB are expressed in human ovarian tissue. Furthermore, a differential qualitative distribution of AGE, RAGE and NF-kappaB p65 subunit was observed in women with PCOS compared to healthy controls, where a stronger localization of both AGE and RAGE was observed in the granulosa cell layer of PCOS ovaries.

The involvement of advanced glycation endproducts (AGEs) in renal injury of diabetic glomerulosclerosis: association with phenotypic change in renal cells and infiltration of immune cells.

Glycation of proteins is regarded as one of the major causes of the progression of diabetic nephropathy (DN) and of the phenotypic changes in glomerular mesangial cells (MC) and the cellular infiltration that occurs in MC from DN. It thus is very important to study the interrelationships and interaction between these causes. The localization of advanced glycation endproducts (AGEs), cytoskeletal proteins, and immune cell infiltration was evaluated by immunohistochemical study in patients with DN. N(Epsilon)-(carboxymethyl)lysine (CML), Alpha-smooth muscle actin (SMA), and low-molecular-weight caldesmon (L-CLD) were localized in the mesangial area in DN. The intensity of mesangial SMA and L-CLD expression was significantly correlated with the index of diabetic glomerulosclerosis (IDGS), while the expression of pentosidine was correlated with the IDGS and with 24-h urinary protein. Pentosidine was localized in glomerular basement membrane (GBM) only in DN and had a significant correlation with the mesangial expression of SMA and L-CLD. Pyrraline deposition on the tubular basement membrane, and the expression of SMA and L-CLD, and the infiltration of immune cells were observed in interstitial areas in DN. The intensity of L-CLD expression had a close relationship with pyrraline deposition and immune cell infiltration. The expression of SMA and L-CLD in interstitial areas was significantly correlated with the percent interstitial volume. AGEs are involved in renal injury in DN, and their effect is, at least in part, exercised via phenotypic changes in intrinsic renal cells and by the infiltration of immune cells.

Extra- and intracellular localization of advanced glycation end-products in human atherosclerotic lesions.

Extra- and intracellular localization of advanced glycation end-products in human atherosclerotic lesions.

Localization of advanced glycation endproducts in the kidney of experimental diabetic rats

Advanced glycation endproducts (AGE) have been proposed as a major mediator in the development of various diabetic complications. In order to evaluate the involvement of AGE in the development of diabetic nephropathy, we examined the localization of AGE in the kidney of the streptozotocin-induced diabetic rats immunohistochemically using a monoclonal antibody directed to AGE. In the diabetic rats, glomerular hypertrophy, thickening of the glomerular basement membrane, and expansion of mesangial matrix were observed. AGE was detected in expanded mesangial area and glomerular basement membrane in the kidneys of diabetic rats. The present results suggest that AGE may participate in the development of diabetic nephropathy.