As a result we examined irrespective of whether CrVI-induced apoptosis of granulosa cells is mediated by way of release of cytochrome c and activation of caspase-3. Final results indicated that CrVI induced release of cytochrome c in the mitochondria into cytosol within a time-dependent method. The increase of cytochrome c protein while in the cytosol was negatively correlated which has a concurrent decrease in the mitochondria whereas vitamin C prevented this translocation . Down-stream of cytochrome c, CrVI elevated cleavage of caspase-3 and PARP proteins. Vitamin C mitigated the effect of CrVI on cleavage of caspase-3 and PARP . CrVI-induced apoptosis of granulosa cells was inhibited by a cytochrome c inhibitor or caspase-3 inhibitor . These benefits together indicate that CrVI induces apoptosis of granulosa cells through cytochrome c and caspase-3 dependent intrinsic apoptotic pathways.
CrVI altered expression of Bcl-2, Bcl-XL, Bax, Poor, HSP70 and HSP90 proteins in granulosa cells The Bcl-2 loved ones proteins constitute crucial parts to regulate activation of intrinsic apoptotic pathways by governing mitochondrialmembrane permeabilization order Nutlin-3 and subsequent release of cytochromec . Moreover to Bcl-2 relatives, heat shock proteins HSP70 or HSP90 are anti-apoptotic by preventing mitochondrial membrane permeabilization . Down-regulation or inhibition of HSP70 or HSP90 protein is sufficient to sensitize a cell for apoptosis . So,we studied the results of CrVI on expression of antiapoptotic proteins Bcl-2 and Bcl-XL, proapoptotic proteins Bax and Lousy, at the same time as HSP70 and HSP90 proteins.
Effects indicated that CrVI decreased expression of Bcl-2 and Bcl-XL proteins, greater complete Negative and Bax proteins, and decreased HSP70 and HSP90 proteins. Vitamin C mitigated results of CrVI on the ranges of Bcl-2, Bcl-XL, Bax and Honokiol Negative, HSP70 and 90 proteins temporally at twelve and 24 h. . From the absence of apoptotic stimuli, Lousy protein is phosphorylated at serine 112 and 136 by MAPK and AKT pathways. Phosphorylated Terrible proteins bind with 14- 3-3 proteins and are sequestered inside the cytosol . Dephosphorylation of Poor is significant for its translocation into antiapoptotic proteins and increases expression of pro-apoptotic proteins in granulosa cells. CrVI altered mitochondrial translocation of BAX, Terrible, HSP70 and HSP90 in granulosa cells Translocation of Bax and Terrible proteins from cytoplasm into mitochondria is critical to execute apoptotic cell death in response to oxidative pressure and DNA injury .
So the effect of CrVI about the translocation of Bax and Lousy was studied. In handle cells, Bcl-2 and Bcl-XL proteins were abundantly localized during the mitochondria, CrVI decreased mitochondrial levels of Bcl-2 and Bcl-XL proteins and vitamin C partially mitigated the effects of CrVI on Bcl-2 but not Bcl-XL protein.