The study of mitochondrial pathway that mediated by oxidative stress involved cardiomyocytes apoptosis induced by N, N-dimethylformamide mitochondrial pathway

Objective To explore whether N, N-dimethylformamide (DMF) can induce apoptosis in H9c2 cardiomyocytes through mitochondrial pathway and its potential mechanism preliminarily. Methods H9c2 Cardiomyocytes were cultured in vitro and divided into groups with different treatments:control group, NAC group (4 mmol/L), DMF group (200 mmol/L), NAC (4 mmol/L)+DMF (200 mmol/L) group. After treating with 24 h, the location of Cyt-c was detected by immunofluorescence cytochemistry (IFC) and the levels of Cyt-c, Bcl-2, Cleaved caspase-3 were detected by western blotting. Results IFC showed that the control and NAC group of Cyt-c were distributed in spotted or patchy shape in the cytoplasm, and the DMF group of Cyt-c was distributed in the cytoplasm diffusely, while NAC could inhibit the effect induced by DMF. Western blotting showed that the levels of Cyt-c different groups were significantly different (F=26.14, P<0.001). Comparing with the control group, as for the level of Cyt-c, the DMF group was significantly increased and the NAC+DMF group was significantly decreased than DMF group (all P<0.001); as for the level of Bcl-2 among different groups were significantly different (F=28.92,P<0.001). Compare with control group, the DMF group was significantly decreased and the NAC+DMF group was significantly increased than DMF group (all P<0.001); as for the level of Cleaved caspase-3, there were significantly different (F=28.98,P<0.001) among different groups. Compare with control group, the DMF group was significantly increased and the NAC+DMF group was significantly decreased than DMF group (all P<0.001). Conclusions DMF can induce apoptosis in H9c2 Cardiomyocytes through mitochondrial pathway. Oxidative stress may be involved in the pathogenesis.