二乙酰吗啡致大鼠小脑颗粒神经元细胞凋亡过程中C-Jun、 Cytc 和Caspase-9 的作用.

BACKGROUND: Diacetylmorphine drugs can cause neuronal injury, but whether diacetylmorphine induces neuronal apoptosis or whether c-Jun, cytc and caspase-9 factors are involved in this process have not been reported. OBJECTIVE: To investigate whether C-jun, Cytc, and Caspase-9 are involved in apoptosis induced by diacetylmorphine. METHODS: The cerebellar granule neurons of SD suckling mice were cultured in vitro for 7 days. Different concentrations of diacetylmorphine (0, 10, 40, 80, 100, 120 mg/L) were used to intervene neurons for 24 hours. The morphological changes of neurons were observed by inverted fluorescence microscope, and the apoptosis rate was detected by flow cytometry. The cells were treated with 100 mg/L diacetylmorphine and 20 μmol/L JNK inhibitor SP600125 for 24 hours. The expressions of c-jun, cytc and caspase-9 protein were detected by immunofluorescence and western blot assay. RESULTS AND CONCLUSION: (1) With the increase dosages of diacetylmorphine, the neuronal cell body of neurons shrank, the brightness decreased, some of them were gray black, the cells were fragmented, the neuron network structure disappeared in different degrees and the number of cell changes increased gradually. (2) With the increase of the concentration of the drug, the number of morphological changes increased obviously (P < 0.01), and the apoptosis rate also increased significantly (P < 0.01). (3) Compared with the control group, c-jun, cytc and caspase-9 proteins were highly expressed in the diacetylmorphine group when treated with 100 mg/L diacetylmorphine and the difference was statistically significant (P < 0.05). Compared with the diacetylmorphine group, c-jun, cytc, and caspase-9 protein expression was significantly down-regulated in the diacetylmorphine+SP600125 group (P < 0.05). (4) The results indicate that diacetylmorphine can induce apoptosis of cerebellar granule neurons. The c-jun, cytc, and caspase-9 are involved in neuronal apoptosis induced by diacetylmorphine. [ABSTRACT FROM AUTHOR]