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Investigating oxidative modification of anti‐oxidant enzymes and protein carbonylation as mechanisms of cigarette smoke‐induced cardiac stem cell damage (732.6)
Author(s) -
Sumanasekera Wasana,
Dao Halle,
Gyamfi Felix,
Shultz Kadi,
Le Uyen,
Rokosh Gregg
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.732.6
Subject(s) - chemistry , catalase , oxidative stress , glutathione , protein carbonylation , superoxide dismutase , antioxidant , biochemistry , nitric oxide , pharmacology , oxidative phosphorylation , western blot , enzyme , medicine , organic chemistry , gene
Rationale and Objective: Cigarette smoking is a major health crisis across the globe resulting in many diseases including cardiovascular injuries. This study mainly focused on delineating mechanisms of CS induced cardiac stem cell (CSC) damage, which was observed earlier by our research group. We hypothesized that cigarette smoke induced reactive oxygen intermediates modify anti oxidant enzymes and proteins present in CSC via carbonylation leading to CSC damage, which can be attenuated by ascorbate treatment. Methods: C‐kit positive CSCs have been used as the experimental model. Following treatment of CSCs with cigarette smoke extract (CSE) for 24 hours, CSE induced oxidative modification of CSC proteins, total glutathione level, catalase activity, inhibition of super oxide dismutase (SOD) activity and ascorbate mediated prevention were assessed. The employed methods were; oxi‐ELISA, oxi‐blot / SDS polyacrylamide electrophoresis followed by western blotting, catalase, SOD activity, and total glutathione detection assays. Results: CSE induced oxidative modification / carbonylation of CSC proteins, inhibition of SOD activity, and alleviation of inhibition by ascorbate was observed. CSE caused increase in both catalase activity and total glutathione level in CSCs, which are indicative of oxidative stress. Ascorbate treatment decreased catalase activity indicating squelching of free radicals. Ascorbate increased total glutathione level in CSCs. Conclusions: Oxidative stress induced modulation of CSC proteins and antioxidant enzyme activity is the main mechanism for CSE induced attenuation of CSC function. Ascorbate served as a preventative measure. Implications: As CS attenuates CSC functions, it is crucial to find a way to optimize CSCs prior to transplant. Ascorbate pre‐conditioning serves as a promising target. Grant Funding Source : Supportyed by Sullivan University Faculty Development Grant