Effects of Inhibiting Iron-Dependent Lipid Peroxidation on Aging
Van Cise, Michelle Lynn
MetadataShow full item record
Aging is an unavoideble process end the mechanisms involved in the cell's demise are still unknown. This study attempted to alter the level of iron-dependent lipid peroxidation in the tissue culture environment by adding known entioxidants to the in vitro cellular environment. Compounds used to accomplish th1s were desferroxamine, vitamin E, U74500A end U74006F. It is proposed that these compounds may act by chelating iron which is required for catalyzation of the ininitiation of lipid peroxidation, except for vitamin E end 74006F which are anti-oxidants whose protective mechanisms involve the scavenging of detrimental species such as the superoxide anion, the hydroxl radical and hydrogen peroxide. Experimentation occurred on three human diploid fibroblastic cell lines: MRC-5, WI 1003, and primary prostatic stroma. Two of these cell types, MRC-5 and WI 1003, were selected because of their derivation from lung tissue, resembling the WI-38 cells used in previous cytogerontological studies. These cells were subjected to concentretions of each drug which were determined to be optimal for the maintenance of cell morphology and fiability. Parallel cultures of each cell type were treated with one of the four drug solutions over their lifespan. The results indicate that MRC-5 and WI 1003 cells treated with desferroxamine increase cell number with each subcultivation. The prostatic stromal cells exhibited a higher cell count with the 74006 drug treatment. In all cases, cells treated w1th drug had higher cell counts than drug-free controls. Experiments designed to reveal differences in DNA synthesis rates and DNA content through triated thymidine pulses and flow cytomentry respectively, showed no correlation with drug treatment. Experiments using epithelial cells to assay for levels of secretion of a marker protein, PSP-15, revealed that cells treated with U74500A in particular, secreted more of the protein than control cells when grown on an extracellular matrix. The results of this study suggest that the addition of an iron chelator to media used in mammalian cell culture may improve cell viability and expression of cellular differentiated function. The tissue culture setting represents an ischemic environment where generation of lipid peroxidation catalyzed by iron can occur. The protective effects of the drugs tested here would warrant similar experiments at the organ culture level to find optimal protective concentrations to use in organ perfusion prior to reimplantation.