Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (25): 3943-3948.doi: 10.12307/2021.002

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C-jun, Cytc and Caspase-9 in the apoptosis of cerebellar granule neurons induced by diacetylmorphine in rats

Su Liping1, Lu Ziyang2, Liu Li2, Zhang Wei1, Su Tianyuan3, Hu Xiayun4, Pu Hongwei5, 6, Han Dengfeng7   

  1. 1Department of Pathology, 3Telemedicine center, 5Discipline Construction Section, 7Department of Neurology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China; 2School of Basic Medicine, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 4Second Department of Thoracic Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China; 6Key Laboratory of Forensic Medicine of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China 
  • Received:2020-06-01 Revised:2020-06-02 Accepted:2020-07-29 Online:2021-09-08 Published:2021-03-24
  • Contact: Pu Hongwei, MD, Professor, Master’s supervisor, Discipline Construction Section, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China; Key Laboratory of Forensic Medicine of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China Han Dengfeng, MD, Chief physician, Master’s supervisor, Department of Neurology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China
  • About author:Su Liping, Master, Technician-in-charge, Department of Pathology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang Uygur Autonomous Region, China Lu Ziyang, Master candidate, School of Basic Medicine, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    the Open Project of Key Laboratory of Forensic Medicine in Hebei Province, No. KF201603 (PHW); the National Natural Science Foundation of China, No. 81860049, No. 81260464 (to PHW)

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Abstract: 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.

Key words: diacetylmorphine, cerebellum, neuron, c-jun, Cytc, Caspase-9, apoptosis

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