Ablation of the gene encoding p66Shc protects mice against AGE-induced glomerulopathy by preventing oxidant-dependent tissue injury and further AGE accumulation.
OBJECTIVE
AGEs have been implicated in renal disease associated with ageing, diabetes and other age-related disorders. Reactive oxygen species (ROS) promote formation of AGEs, which cause AGE-receptor-mediated ROS generation with activation of signalling pathways leading to tissue injury and further AGE accumulation. ROS generation is regulated by the Src homology 2 domain-containing transforming protein C1 (Shc1) isoform p66(Shc), whose deletion has been shown to protect from tissue injury induced by ageing, diabetes, hyperlipidaemia and ischaemia-reperfusion by preventing oxidative stress. This study was aimed at assessing the role of p66(Shc) in the modulation of oxidative stress and oxidant-dependent renal injury induced by AGEs.


METHODS
For 10 weeks, male p66 (shc) knockout (KO) and wild-type (WT) mice were injected with 60 microg/day albumin modified or unmodified by N epsilon-(carboxymethyl) lysine (CML). Mice were then killed for the assessment of renal function and structure, as well as systemic and renal tissue oxidative stress.


RESULTS
Upon CML injection, KO mice, in contrast to WT mice, showed no or only mild forms of proteinuria, glomerular hypertrophy, mesangial expansion, glomerular sclerosis, renal/glomerular cell apoptosis and extracellular matrix upregulation. Moreover, KO mice had lower circulating and tissue AGEs than WT mice and unchanged plasma isoprostane 8-epi-prostaglandin-F(2alpha) levels, renal/glomerular CML, 4-hydroxy-2-nonenal, AGE receptor and NAD(P)H oxidase 4 (NOX4) content (and expression of the corresponding genes), and nuclear factor kappaB activation (NFkappaB). Mesangial cells from KO mice exposed to CML showed no or slight increase in ROS levels and NFkappaB activation, again at variance with WT cells.


CONCLUSIONS
These data indicate that p66(Shc) participates in the pathogenesis of AGE-dependent glomerulopathy by mediating AGE-induced tissue injury and further AGE formation through ROS-dependent mechanisms involving NFkappaB activation and upregulation of Nox4 expression and NOX4 production.
Menini
S
S
Department of Clinical Sciences, 'La Sapienza' University, Viale del Policlinico, 155-00161, Rome, Italy.
Iacobini
C
C
Department of Clinical Sciences, 'La Sapienza' University, Viale del Policlinico, 155-00161, Rome, Italy.
Ricci
C
C
Department of Clinical Sciences, 'La Sapienza' University, Viale del Policlinico, 155-00161, Rome, Italy.
Oddi
G
G
Department of Clinical Sciences, 'La Sapienza' University, Viale del Policlinico, 155-00161, Rome, Italy.
Pesce
C
C
DISTBIMO, University of Genoa Medical School, Genoa, Italy.
Pugliese
F
F
Department of Clinical Sciences, 'La Sapienza' University, Viale del Policlinico, 155-00161, Rome, Italy.
Block
K
K
Department of Medicine, Division of Nephrology, University of Texas Health Science Center, San Antonio, TX, USA.
Abboud
H E
HE
Department of Medicine, Division of Nephrology, University of Texas Health Science Center, San Antonio, TX, USA.
Giorgio
M
M
Department of Experimental Oncology, European Institute of Oncology, Milan, Italy.
Migliaccio
E
E
Department of Experimental Oncology, European Institute of Oncology, Milan, Italy.
Pelicci
P G
PG
Department of Experimental Oncology, European Institute of Oncology, Milan, Italy.
Pugliese
G
G
Department of Clinical Sciences, 'La Sapienza' University, Viale del Policlinico, 155-00161, Rome, Italy. giuseppe.pugliese@uniroma1.it.
D048868:Adaptor Proteins, Signal Transducing / Q000172:deficiency* / Q000235:genetics*; D000818:Animals; D017931:DNA Primers; D005838:Genotype; D005920:Glomerular Mesangium / Q000473:pathology*; D007150:Immunohistochemistry; D007674:Kidney Diseases / Q000473:pathology* / Q000517:prevention & control*; D007678:Kidney Glomerulus / Q000473:pathology* / Q000503:physiopathology; D051379:Mice; D008815:Mice, Inbred Strains; D016328:NF-kappa B / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000502:physiology*; D055765:Shc Signaling Adaptor Proteins; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1
D048868:Adaptor Proteins, Signal Transducing; D017931:DNA Primers; D016328:NF-kappa B; D017382:Reactive Oxygen Species; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D055765:Shc Signaling Adaptor Proteins; C528034:Shc1 protein, mouse; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1