Therefore, we aim to investigate the

role of Sirt1 in diabetic nephropathy (DN). Methods and Results: We found that Sirt1 in proximal tubules (PTs) was downregulated before albuminuria, and, thereafter, Sirt1 in podocytes (Pods) was downregulated in DN mice including both streptozotocin-induced and obese (db/db) mice. Then, we created PT-specific Sirt1 transgenic (Tg) and conditional knockout (CKO) mice to examine the role of PT’s Sirt1. Sirt1 Tg prevented and CKO aggravated glomerular changes and albuminuria that occured in diabetes, respectively. Non-diabetic CKO mice exhibited albuminuria, suggesting that Sirt1 in PTs affects glomerular function. We also observed that reduced PT’s Sirt1 in DN decreased selleck chemical NMN (Nicotinamide Mono Nucleotide, a key intermediate of Sirt1-related nicotinic acid metabolism) led to decreasing Pod’s Sirt1. Reduced Sirt1 increased Claudin-1, a tight junction protein, in Pods by an epigenetic mechanism whereby decreased Pod’s Sirt1 inactivated

Dnmt1 leading to reduced CpG methylation of Claudin-1 gene, which contributed to increased Claudin-1 expression and albuminuria. Intriguingly, Claudins are generally known to strengthen the epithelial barrier, but we novely showed that overexpression of Claudin-1 in Pods increased glomerular permeability by activating β-catenin–Snail pathway. We also demonstrated retrograde interplay from PTs to Pods mediated by NMN by analyzing Hydroxychloroquine conditioned RG7420 medium experiments, measurement of renal endogenous NMN and injection of fluorescence-labeled exogenous NMN. In human renal biopsy with DN, the levels of decreased Sirt1 in PT or Pods and increased Claudin-1 in Pods were correlated

with proteinuria levels. Conclusion: Our results (Hasegawa K, Nature Medicine 2013) suggest that Sirt1 in PTs protects against diabetic albuminuria by maintaining NMN around Pods, thus influencing glomerular function. Although tubulo-glomerular feedback has been previously reported, ours is the first description of a proximal tubular substance (NMN) that communicates with podocytes as a key mediator of intracellular crosstalk. GALLO LINDA A.1, WARD MICHEAL S.1, HARCOURT BROOKE E.1, FOTHERINGHAM AMELIA K.1, MCCARTHY DOMENICA A.1, PENFOLD SALLY A.2, FORBES JOSEPHINE M.1,3 1Glycation and Diabetes, Mater Research Institute-UQ, Australia; 2Diabetes Complications Division, Baker IDI Heart and Diabetes Institute, Australia; 3School of Medicine, Mater Clinical School, University of Queensland, Australia Introduction: The plasma concentration of the reactive carbonyl, methylglyoxal (MGO), is elevated in diabetes. Increased accumulation of MGO may contribute to insulin resistance at peripheral sites of glucose uptake. A deficiency in podocyte insulin signalling impairs podocyte function resulting in kidney disease. Glyoxalase-1 (GLO-1) is an enzyme considered to detoxify MGO. Hence, we examined the effects of inhibiting GLO-1 on podocyte insulin signalling and renal function under diabetic conditions.