Impaired retinal angiogenesis in diabetes: role of advanced glycation end products and galectin-3

Suppression of angiogenesis during diabetes is a recognized phenomenon but is less appreciated within the context of diabetic retinopathy. The current study has investigated regulation of retinal angiogenesis by diabetic serum and determined if advanced glycation end products (AGEs) could modulate this response, possibly via AGE-receptor interactions. A novel in vitro model of retinal angiogenesis was developed and the ability of diabetic sera to regulate this process was quantified. AGE-modified serum albumin was prepared according to a range of protocols, and these were also analyzed along with neutralization of the AGE receptors galectin-3 and RAGE. Retinal ischemia and neovascularization were also studied in a murine model of oxygen-induced proliferative retinopathy (OIR) in wild-type and galectin-3 knockout mice (gal3(-/-)) after perfusion of preformed AGEs. Serum from nondiabetic patients showed significantly more angiogenic potential than diabetic serum (P <0.0001) and within the diabetic group, poor glycemic control resulted in more AGEs but less angiogenic potential than tight control (P <0.01). AGE-modified albumin caused a dose-dependent inhibition of angiogenesis (P <0.001), and AGE receptor neutralization significantly reversed the AGE-mediated suppression of angiogenesis (P <0.01). AGE-treated wild-type mice showed a significant increase in inner retinal ischemia and a reduction in neovascularization compared with non-AGE controls (P <0.001). However, ablation of galectin-3 abolished the AGE-mediated increase in retinal ischemia and restored the neovascular response to that seen in controls. The data suggest a significant suppression of angiogenesis by the retinal microvasculature during diabetes and implicate AGEs and AGE-receptor interactions in its causation.

Advanced Glycation End Products (AGEs) accumulate in the Reproductive Tract of Men with Diabetes.

Light microscopic studies comparing sperm parameters show little association between diabetes and male fertility. However, with the introduction of new analytical techniques, evidence is now emerging of previously undetectable affects of diabetes on sperm function. Specifically, a recent study has found significantly higher sperm nuclear DNA (nDNA) fragmentation in diabetic men. As advanced glycation end products (AGEs) are important instigators of oxidative stress and cell dysfunction in numerous diabetic complications, we hypothesized that these compounds could also be present in the male reproductive tract. The presence and localization of the most prominent AGE, carboxymethyl-lysine (CML), in the human testis, epididymis and sperm was determined by immunohistochemistry. Parallel ELISA and Western blot analyses were performed to ascertain the amount of CML in seminal plasma and sperm from 13 diabetic and 9 non-diabetic subjects. CML immunoreactivity was found through out the seminiferous epithelium, the nuclei of spermatogonia and spermatocytes, in the basal and principle cells (cytoplasm and nuclei) of the caput epididymis and on most sperm tails, mid pieces and all cytoplasmic droplets. The acrosomal cap, especially the equatorial band, was prominently stained in diabetic samples only. The amount of CML was significantly higher (p = 0.004) in sperm from non diabetic men. Considering the known detrimental actions of AGEs in other organs, the presence, location and quantity of CML, particularly the increased expression found in diabetic men, suggests that these compounds may play a hitherto unrecognized role in male infertility.

Homocysteine is lower in the Third Trimester of Pregnancy in Women with Enhanced Folate status from continued Folic Acid Supplementation.

Background: In many countries current recommendations are that women take a daily 400ug folic acid supplement, from before conception until the end of the 12th week of gestation, for the prevention of neural tube defects. Low folate status is associated with an elevated concentration of plasma total homocysteine (tHcy), a risk factor that is associated with pregnancy complications such as pre-eclampsia. Methods: In a longitudinal study, tHcy and corresponding folate status were determined in 101 pregnant women at 12, 20 and 35 weeks of gestation, in 35 non-pregnant control subjects sampled conconcurrently, and in a subgroup (n=21 pregnant, 19 non-pregnant women) at 3 days post-partum. Results: Plasma tHcy concentrations were significantly lower throughout pregnancy compared with control subjects, with values lowest in the 2nd trimester before increasing toward non-pregnant values in the 3rd trimester. Importantly, tHcy concentrations were lower in pregnant women taking folic acid supplements compared to those not, an effect which reached significance in the 3rd trimester (5.25 umol/l v 6.89 umol/l, P <0.05). Furthermore, during the 3rd trimester, tHcy concentrations were significantly higher in pregnant women with a history of miscarriage compared to those with no previous history (7.32 umol/l v 5.62 u­mol/l, P <0.01). Conclusion: This is the first longitudinal study to show that homocysteine levels rise in late pregnancy towards non-pregnant levels; a rise which can be limited by enhancing folate status through continued folic acid supplementation. These results indicate a potential role for continued folic acid supplementation in reducing pregnancy complications associated with hyperhomocysteinaemia.