Journal
NEUROCHEMICAL RESEARCH
Volume 31, Issue 8, Pages 1011-1020Publisher
SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s11064-006-9107-x
Keywords
bone marrow mesenchymal stem cells; heat shock proteins; oxidative stress
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The phenomenon of neuronal transdifferentiation performed on bone marrow mesenchymal stem cells (MSCs) has been criticized by recent studies indicating that acquired neuron-like morphology of induced MSCs is caused by cellular stress. Therefore, to test this hypothesis we have investigated whether exposure of rat MSCs (rMSCs) to chemical inducer 2 mM beta-mercaptoethanol (BME) for 1-3 h followed by 24 h incubation leads to HSP72 synthesis, thus suggesting higher resistance of rMSCs to oxidative damage. Present data from immunohistochemistry clearly indicate development of time-dependent subcellular HSP72 distribution, initially seen in nuclei at 1 h followed by its translocation to surrounding central cytoplasm and processes at 2-3 h after BME stimulation. Western blot (WB) analysis confirmed the expression of HSP72 protein in induced rMSCs at both stimulation periods. Furthermore, preconditioned rMSCs with BME for 1 h expressing HSP72 positivity at 24 h showed higher resistance ( 78 +/- 10% of survival cells) to oxidative stress caused by 1 mM H2O2 when compared to those preconditioned for 3 h ( 59 +/- 8% of survival cells) or control-unconditioned rMSCs exposed to the same stressor conditions ( 56 +/- 6% of survival cells). Thus, the cellular protection was lost if the duration of BME preconditioning was increased as far as possible ( 3 h) ( while still remaining sub-lethal). This suggests that exposure of rMSCs to the optimal concentration of BME ( 2 mM) during optimal induction period ( 1 h) mediate their protection and increases resistance to oxidative injury, while over crossing these limits is in-effective. In addition, our findings confirm that cultured rMSCs remain competent to be preconditioned by BME, through a pathway that may increase the antioxidant balance or involve activation of HSP72 protein induced tolerance.
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