In this context, it had been recommended that mitochondrial membrane permeability transition is usually a possible mechanism of diclofenac induced apoptosis in hepatocytes, and it had been observed that diclofenac induced MPT in isolated rat liver mitochondria and in key cultured hepatocytes . Lately, it’s been reported that reactive oxygen species are concerned in diclofenac induced apoptosis of cultured gastric cells and nephrotoxicity in vivo and that oxidative damage in the mitochondrial degree is involved in MPT induction in diclofenac treated hepatocytes . The mechanisms underlying the induction of apoptosis by diclofenac in hepatocytes, gastric cells, and kidney are reported, but the causal sequence and molecular mechanism of diclofenac induced apoptosis in leukocytes haven’t been entirely elucidated. The elucidation with the diclofenac induced apoptotic mechanism might be significant for avoiding its unwanted effects or applying diclofenac for the treatment method of human cancer.
Within the basis of those data, we studied the mechanism of diclofenac induced apoptosis of promyelocytic leukemia, HL cells, and the usefulness of diclofenac for antileukemic therapy. Diclofenac inhibited the growth of HL cells within a concentration dependent method. Total suppression was observed at concentrations higher than AM at h . Diclofenac induced characteristic morphological modifications Maraviroc price of apoptosis but not necrosis, this kind of as cell shrinkage and nuclear fragmentation as proven by staining cells with Hoechst and propidium iodide . The DNA content material of cells was quantified by flow cytometry of propidium iodide stained cells, which also permitted evaluation of which phase from the cell cycle the cells had been in along with the quantity of apoptotic cells. We noticed that diclofenac was able to induce cell cycle arrest at the G M checkpoint, thereby preventing cell division and triggering cells to get a G DNA written content . Diclofenac induces DNA fragmentation, which can be suppressed by z VAD fmk and Ac IETD CHO, but not by CsA Diclofenac induced DNA fragmentation of HL cells within a time and concentration dependent method .
Obvious DNA fragmentation was observed at concentrations higher than AM at h, TGF-beta inhibitor and time course examination exposed that DNA fragmentation grew to become obvious h following incubation with AM diclofenac. As MPT and caspases perform important roles inside the operation of apoptosis, we investigated the effects of CsA, a specific inhibitor of MPT, and z VAD fmk, a universal caspase inhibitor, on diclofenac induced apoptosis of HL cells. The diclofenac induced DNA fragmentation was strongly suppressed by AM z VAD fmk, but not by AM CsA. Moreover, diclofenac induced DNA fragmentation was suppressed by pretreatment with AM Ac IETD CHO, a caspase inhibitor, exhibiting the involvement of caspase in the system of apoptosis .