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Background: The presence of accelerated atherosclerosis in patients with kidney disease cannot be entirely explained by traditional cardiovascular risk factors. Exposure to urea, which is normally present in human blood plasma and elevated in patients with kidney disease, leads to the carbamylation of proteins. We postulated that low-density lipoprotein (LDL) carbamylated by urea has biologic effects relevant to atherosclerosis., Methods: To produce carbamylated LDL (cLDL), human native LDL (nLDL) was chemically modified in vitro by exposure to potassium cyanate. Human coronary artery endothelial cells (HCAECs) and human coronary artery smooth muscle cells (CASMCs) were treated in vitro with cLDL or nLDL. Irreversible cell death was measured using the lactate dehydrogenase (LDH) assay, apoptosis was assessed by annexin V binding, and proliferation was determined using bromodeoxyuridine (BrdU) incorporation. Total plasma protein carbamylation and plasma cLDL were measured in hemodialysis patients using the homocitrulline assay and enzyme-linked immunosorbent assay (ELISA)., Results: Our studies demonstrated that cLDL but not nLDL induced dose-dependent vascular cell injuries relevant to atherosclerosis, which included the proliferation of vascular smooth muscle cells and endothelial cell death. Under light microscopy, endothelial cells treated with cLDL showed signs of morphologic alterations. The injury to endothelial cells measured by LDH release was time-dependent and correlated with the degree of LDL carbamylation. At least a part of the endothelial cell population treated with cLDL died by apoptosis. In patients with advanced renal disease on hemodialysis, total plasma protein carbamylation and plasma cLDL were several times higher than in control healthy individuals., Conclusion: Collectively these data suggest the potential role of carbamylated LDL in accelerated atherosclerosis in patients with chronic renal disease and, possibly, in healthy individuals.

Atherogenic lipoproteins and reactive oxygen species stimulate renin release from isolated juxtaglomerular (JG) cells. Here we assessed whether stimulation of renin release is mediated by formation of superoxide anion (O2-), and whether the effects of oxidized lipoproteins, like in many other biological systems, can be prevented by the antiatherogenic high density lipoprotein (HDL). Lipoproteins were prepared from human plasma, and JG cells from mouse and rat kidneys. Basal renal activity of JG cells was measured in culture supernatants and cells, and was dose-dependently and significantly stimulated by oxidized LDL (50 and 300 micrograms/ml) and by oxidized Lp(a) (1, 10 and 30 micrograms/ml). Administration of HDL alone had no effect on renin release. However, coincubation with 100 micrograms/ml HDL significantly suppressed oxidized LDL- and oxidized Lp(a)-stimulated renin release. O2- production of JG cells was directly measured using a chemiluminescence assay. Stimulation with 10 micrograms/ml oxidized LDL and oxidized Lp(a) significantly increased the O2- generation of JG cells. In the presence of 5 micrograms/mL HDL, O2- production was reduced to control levels. These data indicate that stimulation of JG cells with oxidized LDL and Lp(a) induces formation of O2-, which may stimulate renin release in an autocrine fashion. Renin release can be prevented by HDL, presumably by preventing the formation of O2-.

Abnormal lipid accumulation in glomeruli is a recognized early event in the development of glomerulosclerosis. The presence of LDL and scavenger receptors has recently been demonstrated in glomerular cells, including the visceral epithelial cells. To explore the possible molecular mechanisms of lipid-induced glomerular injury, the present investigation was conducted to examine the effects of oxidized LDL (ox-LDL) on the expression of transforming growth factor (TGF)-beta and fibronectin by cultured human glomerular epithelial cells (GEC). Cultured GEC were exposed to human ox-LDL (0 to 100 micrograms/ml) for various time points. Ox-LDL induced a dose- and time-dependent increase in the expression of TGF-beta mRNA. Actinomycin D, a transcriptional inhibitor, but not cycloheximide, a protein synthesis inhibitor, inhibited the response. GEC exposed to ox-LDL also demonstrated elevated levels of fibronectin mRNA. In addition, treatment of GEC with ox-LDL resulted in increased TGF-beta and fibronectin protein expression as detected by immunocytochemistry. Addition of anti-TGF-beta antibody significantly inhibited the increase in fibronectin message level induced by ox-LDL. These data suggest that ox-LDL stimulates matrix protein fibronectin in GEC by a mechanism involving expression of TGF-beta. Thus, accumulation of lipids in human glomerular epithelial cells may contribute to the pathogenesis of glomerulosclerosis through TGF-beta mediated mechanism(s).