Apoptosis in HepG2 cells exposed to high glucose.
Hyperglycemia which characterizes diabetes, leads to several abnormalities in the cellular pathways. We examined the toxicity of glucose in human hepatoma HepG2 cells. HepG2 cells when incubated with 50mM glucose for 72h showed altered morphology i.e. presence of detached and shrunken rounded cells. Glucose treated HepG2 cells also exhibited a significant decrease in viability. Caspase-3 activity and Annexin V staining were significantly increased in glucose treated HepG2 cells, suggesting an apoptotic mode of cell death. Glucose induced apoptosis in HepG2 cells was a consequence of increased oxidative stress as evidenced by the increased reactive oxygen species (ROS) level, lipid peroxidation, protein carbonyl and 3-nitrotyrosine adduct formation. The intracellular antioxidant glutathione was found to be increased in HepG2 cells treated with glucose, possibly to aid the cells to overcome the persistent oxidative stress elicited by glucose in HepG2 cells. N-Acetyl cysteine, a precursor of glutathione and an antioxidant was effective in reversing the morphological changes, increasing the viability, decreasing the ROS level and 4-hydroxynonenal and 3-nitrotyrosine adduct formation, thus validating the role of oxidative stress as a major mechanism for glucose induced apoptosis in HepG2 cells. These results suggest that glucose induces apoptosis in liver cells through increased oxidative stress.
Chandrasekaran
Karthikeyan
K
Life Science Division, AU-KBC Research Centre, MIT Campus of Anna University, Chromepet, Chennai 600 044, India.
Swaminathan
Kavitha
K
Chatterjee
Suvro
S
Dey
Aparajita
A
D017302:Annexins / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005947:Glucose / Q000008:administration & dosage / Q000633:toxicity*; D006801:Humans; D007770:L-Lactate Dehydrogenase; D008113:Liver Neoplasms; D008353:Mannitol / Q000494:pharmacology; D013194:Staining and Labeling; D013997:Time Factors
D017302:Annexins; D008353:Mannitol; D007770:L-Lactate Dehydrogenase; D053148:Caspase 3; D005947:Glucose