Inherent differences in morphology, proliferation, and migration in saphenous vein smooth muscle cells cultured from nondiabetic and Type 2 diabetic patients

Abstract

Madi HA, Riches K, Warburton P, O'Regan DJ, Turner NA, Porter KE. Inherent differences in morphology, proliferation, and migration in saphenous vein smooth muscle cells cultured from nondiabetic and Type 2 diabetic patients. Am J Physiol Cell Physiol 297: C1307-C1317, 2009. First published September 9, 2009 doi:10.1152/ajpcell.00608.2008.-Individuals with Type 2 diabetes mellitus (T2DM) are at increased risk of saphenous vein (SV) graft stenosis following coronary artery bypass. Graft stenosis is caused by intimal hyperplasia, a pathology characterized by smooth muscle cell (SMC) proliferation and migration. We hypothesized that SV-SMC from T2DM patients were intrinsically more proliferative and migratory than those from nondiabetic individuals. SV-SMC were cultured from nondiabetic and T2DM patients. Cell morphology (light microscopy, immunocytochemistry), S100A4 expression (real-time RT-PCR, immunoblotting), proliferation (cell counting), migration (Boyden chamber assay), and cell signaling (immunoblotting with phosphorylation state-specific antibodies) were studied. SV-SMC from T2DM patients were morphologically distinct from nondiabetic patients and exhibited a predominantly rhomboid phenotype, accompanied by disrupted F-actin cytoskeleton, disorganized ␣-smooth muscle actin network, and increased focal adhesion formation. However, no differences were observed in expression of the calcium-binding protein S100A4, a marker of rhomboid SMC phenotype, between the two cell populations. T2DM cells were less proliferative in response to fetal calf serum than nondiabetic cells, but both populations had similar proliferative responses to insulin plus PDGF. Under high glucose concentration conditions in the presence of insulin, migration of diabetic SV-SMC was greater than nondiabetic cells. Glucose concentration did not affect SV-SMC proliferation. No differences in insulin or PDGF-induced phosphorylation of ERK-1/2 or components of the Akt pathway (Akt-Ser473, Akt-Thr308, and GSK-3␤) were apparent between the two populations. In conclusion, SV-SMC from T2DM patients differ from nondiabetic SV-SMC in that they exhibit a rhomboid phenotype and are more migratory, but less proliferative, in response to serum. diabetes mellitus; vein graft stenosis; metabolic memory DIABETES MELLITUS IS A SERIOUS and escalating health problem worldwide (3) that currently affects more than 2

Deficient leukemia inhibitory factor signaling in muscle precursor cells from patients with type 2 diabetes

“…A reduced proliferative capacity has been demonstrated in smooth muscle cells isolated from patients with type 2 diabetes (24). In this study, we show that LIF-induced cell proliferation is impaired in myoblasts from patients with type 2 diabetes.…”

Progressive Development of Aberrant Smooth Muscle Cell Phenotype in Abdominal Aortic Aneurysm Disease

“…In the current study, we have extended our morphological characterisation by noting marked aberrations in the F-actin cytoskeleton. Similar F-actin features - i.e., truncated and disorganised fibres - have also been described in SV-SMC from patients with type 2 diabetes (T2DM) [20,21,22]. As both aneurysm disease and T2DM are considered to be conditions characterised by premature aging [23,24], it is feasible that disorganisation of F-actin in SMC is a feature of aged cells rather than representative of a particular disease pathology per se.…”

Cellular and molecular mechanisms of endothelial dysfunction in diabetes

“…For 88 Liu et al, 89 Li et al, 90 Dentelli et al 93 ↓ this reason, it is apparent that identifying the presence of a diabetic metabolic memory might be most accurately observed in EC derived from human diabetic vessels. In such a way, we have previously identified inherent functional differences between vascular SMC cultured from patients with and without T2DM, 107 such data offering an explanation for increased venous bypass graft stenosis after revascularisation surgery in these patients. Identification of any sustained functional differences in T2DM-EC may provide further justification for the aforementioned graft failure.…”