Role of Bax Protein and Caspase-3 at High Glucose-Induced Apoptosis in Human Embryonic Kidney (HEK) 293 Cells

Abstract

Background : Hyperglycemia is the principal factor responsible for microvascular complications of diabetes. Diabetic nephropathy is a serious and common complication leading to end stage renal disease. The exact molecular mechanisms of high glucose-induced toxicity on renal cells are still incompletely understood. Therefore in the present study, glucose-induced toxicity was studied in HEK (human embryonic kidney) cells as an in vitro model for diabetic nephropathy.Materials and Methods : First, the viability of HEK 293 cells exposed to glucose was measured by MTT (Methyl Thiazolyl Tetra-zolium) assay. Caspase-3 activity was determined spectrophotometrically using enzyme specific substrate. Moreover, the alteration in expression of Bax, Bcl2 and caspase-3 were measured by Western blotting.Results : The results showed that high glucose significantly reduced cell viability after 48 h (p