Studying the effect of LPS on cytotoxicity and apoptosis in PC12 neuronal cells: role of Bax, Bcl-2, and Caspase-3 protein expression.
Gram-negative bacteria and their endotoxins may be a causal or complicating factor in many serious diseases. Bacterial lipopolysaccharides (LPS) could potentially be human pathogens. Among the many disorders induced by LPS, neurodegenerative diseases such as Parkinson's are reported and are of great interest. Despite the evidence on LPS-induced neurodegeneration, the exact mechanism is unknown. The purpose of this study was to investigate the cytotoxic effect of LPS and also to examine the involvement of Bax, pro-apoptotic, Bcl-2, anti-apoptotic, and caspase-3, the executioner of apoptosis, protein expression, during LPS-induced apoptosis in neuronal PC12 cells. The cell viability was evaluated by MTT assy. The expression of pro-apoptotic Bax and anti-apoptotic Bcl2 and caspase -3 protein expressions were measured by immunoblotting. The results showed that LPS could reduce cell viability after 72 h in a dose-dependent manner which was statistically significant in concentration of 200 microg/ml. In western blotting analysis, LPS (200 microg/ml) also enhanced expression of pro-apoptotic Bax and pro-caspase-3 proteins compared to controls, while the expression of Bcl-2 protein was not changed significantly. The Bax/Bcl-2 ratio was significantly increased in LPS-treated cells compared to controls. From the present results, it might be concluded that LPS can cause PC12 cell death, in which apoptosis plays an important role, possibly by the mitochondrial pathway through higher expression of the Bax as well as caspase 3 protein.
Razi Institute for Drug Research, Iran University of Medical Sciences, Tehran, Iran. sharifal@yahoo.com
Ali M
AM
Sharifi
Fotovat Eskandari
FE
Hoda
Amir Mozaffari
AM
Noor
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D053148:Caspase 3 / Q000096:biosynthesis*; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D008070:Lipopolysaccharides / Q000633:toxicity*; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D016716:PC12 Cells; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis*; D051381:Rats; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis*
C494828:Bax protein, rat; D008070:Lipopolysaccharides; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; C440499:lipopolysaccharide, Escherichia coli O111 B4; C505410:Casp3 protein, rat; D053148:Caspase 3