Up-regulation of 14-3-3β plays a role in intimal hyperplasia following carotid artery injury in diabetic Sprague Dawley rats by promoting endothelial cell migration and proliferation.

PURPOSE

The objective of this study was to determine whether diabetes mellitus (DM)-induced up-regulation of 14-3-3β (YWHAB) in endothelial cells enhances intimal hyperplasia in carotid artery-injured DM Sprague-Dawley (SD) rats.

METHODS

YWHAB expression and rat aortic endothelial cell (RAOEC) vitality were examined using Cell Counting Kit-8 (CCK-8), quantitative reverse transcription PCR (qRT-PCR), and western blot analysis in cells treated with different glucose concentrations (5.6, 10, 15, 25, or 35 mM). For in vivo experiments, a YWHAB small interfering (si) RNA recombinant lentiviral vector (YWHAB-LV) or Mock siRNA recombinant lentiviral vector (Mock-LV) were injected into streptozotocin-induced DM SD rats via the tail vein. YWHAB expression and carotid artery morphology were assessed 7 days post injury using immunofluorescence (IF) and hematoxylin-eosin (HE) staining. The proliferation and migration of Mock-LV and YWHAB-LV-infected RAOECs treated with 25 mM glucose were examined using cell scratch tests and flow cytometry. BCL2-Associated X (BAX) distribution in RAOECs treated with 25 mM glucose was examined using IF staining and western blot analysis.

RESULTS

Western blot, qRT-PCR, and CCK-8 analyses demonstrated that both YWHAB expression and cell vitality increased with increasing glucose concentration (p <0.05). YWHAB IF staining was increased in DM rats compared with the normal group (p <0.05). HE staining showed that intimal hyperplasia is alleviated in YWHAB-silenced DM rats (p <0.05). YWHAB silencing suppressed the proliferation and migration of RAOECs treated with 25 mM glucose (p <0.05). Moreover, western blot analyses and IF staining demonstrated that YWHAB silencing increased the translocation of BAX from the cytoplasm to mitochondria in RAOECs treated with 25 mM glucose (p <0.05).

CONCLUSIONS

Our results indicate that hyperglycemia-induced up-regulation of YWHAB in endothelial cell plays a significant role in intimal hyperplasia following carotid artery injury by enhancing endothelial cell proliferation and migration. YWHAB inhibition in hyperglycemic patients may constitute a potential target for therapeutic interventions via restenosis prevention.