Publications (2)6.88 Total impact
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ABSTRACT: Proteomic analysis is defined as the characterization of the entire set of proteins encoded by a genome. Two-dimensional (2D) electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) are key technologies used in proteomic analysis to gain information about protein expression profiles and post-translational modifications. Knowledge about aging processes can be gained by recognizing changes in protein expression. Thus, to better understand the aging process through protein profiling, post-mitochondrial (PM) fractions of young (13-month) and old (31-month) male Fischer 344 rat kidney were differentially analyzed by 2D. We detected a total number of 380 spots on 2D gel images. Among them, 167 spots showed 2-fold significant alterations (p<0.05) between young and old PM fractions. Further, 103 proteins were identified by MALDI-TOF MS. The PM fraction of aged rat kidney showed increases in antioxidative and proteolytic proteins and decreases in cytoskeletal proteins. In addition, we found age-related changes in transport and homeostasis proteins. Thus, our results demonstrated that proteomic analysis can be effectively applied to the assessment of the age status of protein expression, and thereby provide valuable information on age-related changes of proteome.
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ABSTRACT: Oxidative stress is thought to be a causative factor for age-related damage in a wide variety of cellular constituents that can lead to dysfunction and various pathological conditions, including the inflammatory process. At the molecular level, the redox-sensitive transcription factor, NF-κB plays a key role in the regulation of the inflammatory process, along with cytokines, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS). We studied the mechanism underlying the modulation of the inflammatory reaction with age by investigating NF-κB activation and the expression of COX-2, iNOS, and cytokines genes in hepatic tissues isolated from young and old rats. We expanded our investigation of these factors in rats injected with the inflammatory activator, lipopolysaccharide (LPS). Data showed that NF-κB activity was up-regulated with age and was further enhanced by LPS injection, indicating an increased susceptibility and sensitivity to the inflammatory stimulus with age. To explore further the molecular events leading to NF-κB activation, we investigated the inhibitory component of NF-κB complex, IκB. Cytosolic IκBα, but not IκBβ, was significantly decreased in both old and LPS-treated rats, signifying the enhanced migration of cytosolic NF-κB complex into the nucleus following dissociation from the inhibitor. The appearance of the polypeptide, p65, as determined in the nucleus, corresponded with the change in IκBα, providing further supporting evidence for the molecular process involved in NF-κB activation. Our additional investigation of two proinflammatory-related enzymes, COX-2 and iNOS, and three cytokines, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α, clearly showed aged-related increases, in corroboration with the NF-κB activation. Our results demonstrated that LPS injection caused the enhanced gene expression of inducible proinflammatory proteins, COX-2 and iNOS through NF-κB activation.
Pusan National University
Pusan, Busan, South Korea
- • Division of Pharmacy
- • College of Pharmacy