Role of the mitochondrial Omi/HtrA2 signaling pathway in neuronal apoptosis after craniocerebral injury

doi:10.3877/cma.j.issn.1674-6880.2018.02.007

Abstract

Abstract:

Objective

To investigate the role of the mitochondrial Omi/HtrA2 signaling pathway in neuronal apoptosis after craniocerebral injury in rats.

Methods

A total of 90 rats were randomly divided into the sham operation group, craniocerebral injury group and UCF-101 intervention group, 30 rats in each group; at 12, 24 and 48 h, each group was further divided into three subgroups at random, 10 rats in each subgroup. The animal model of free-fall brain injury in rats was used. Rats in the UCF-101 intervention group were intraperitoneally injected with UCF-101 at a dose of 1.5 μmol/kg 0.5 h before the formation of the model, and rats in the other two groups were intraperitoneally injected with 1 mL sodium chloride solution. At 12 h, 24 h and 48 h, 10 rats in each group were sacrificed to isolate the hippocampal tissues. Terminal deoxynucleotidyl transferase-mediated dUTP Nick end labeling (TUNEL) assay was used to detect neuronal apoptosis in hippocampal tissues of rats in each group, the Western-blotting method was used to detect the expressions of Omi/HtrA2, X-linked inhibitor of apoptosis protein (XIAP), pro-caspase-3, pro-caspase-9 and shearing poly-ADP-ribose polymerase (PARP), and the tetrapeptide fluorescence substrate method was used to detect the activity of caspase-3 and caspase-9.

Results

The ratio of apoptotic hippocampal neurons, the expressions of Omi/HtrA2, pro-caspase-3, pro-caspase-9, shearing PARP and XIAP, and the activity of caspase-3 and caspase-9 were all statistically significantly different in the three groups (F=217.742, 107.139, 145.748, 133.970, 103.029, 65.112, 142.772, 96.187; all P < 0.05). At 12 h, 24 h and 48 h, the ratio of apoptotic hippocampal neurons, the expressions of Omi/HtrA2, pro-caspase-3, pro-caspase-9 and shearing PARP, and the activity of caspase-3 and caspase-9 were significantly higher in the craniocerebral injury group than in the sham operation group (all P < 0.05), while they were significantly lower in the UCF-101 intervention group than in the craniocerebral injury group (all P < 0.05). The expressions of XIAP in hippocampi in the craniocerebral injury group at different time points were significantly lower as compared to the sham operation group (all P < 0.05), while the expressions of XIAP in the UCF-101 intervention group were significantly higher as compared to the craniocerebral injury group (all P < 0.05).

Conclusion

The mitochondrial pathway mediated by Omi/HtrA2 may be involved in the process of neuronal apoptosis after craniocerebral injury in rats, and UCF-101 can effectively inhibit the apoptosis of nerve cells.

Key words: Rat, Craniocerebral injury, Neuronal apoptosis, Omi/HtraA2, UCF-101

Fanghui Chen, Yi Wang, Xiaoqiao Dong, Yun Xie, Zixi Chen, Chen Xiao, Xue Zhao, Weihong Zhu. Role of the mitochondrial Omi/HtrA2 signaling pathway in neuronal apoptosis after craniocerebral injury[J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2018, 11(02): 104-109.

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References 27

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组别	例数	12 h	24 h	48 h
假手术组	30	10.1 ± 0.6^a	9.7 ± 0.6^a	10.5 ± 0.8^a
颅脑损伤组	30	32.2 ± 1.5	50.0 ± 2.0	45.4 ± 2.5
UCF-101干预组	30	25.7 ± 1.1^a	39.0 ± 1.4^a	33.5 ± 1.5^a

组别	例数	12 h	24 h	48 h
假手术组	30	10.1 ± 0.6^a	9.7 ± 0.6^a	10.5 ± 0.8^a
颅脑损伤组	30	32.2 ± 1.5	50.0 ± 2.0	45.4 ± 2.5
UCF-101干预组	30	25.7 ± 1.1^a	39.0 ± 1.4^a	33.5 ± 1.5^a

组别	例数	Omi/HtrA2			pro-caspase-3			pro-caspase-9
组别	例数	12 h	24 h	48 h	12 h	24 h	48 h	12 h	24 h	48 h
假手术组	30	100.0 ± 1.6^a	101.1 ± 1.5^a	101.5 ± 1.8^a	100 ± 6^a	100 ± 5^a	109 ± 5^a	100 ± 5^a	105 ± 4^a	107 ± 5^a
颅脑损伤组	30	165.6 ± 8.0	218.8 ± 8.8	204.0 ± 8.1	177 ± 4	239 ± 10	203 ± 8	182 ± 10	267 ± 13	204 ± 9
UCF-101干预组	30	129.9 ± 4.7^a	170.3 ± 6.8^a	151.7 ± 8.4^a	157 ± 5^a	193 ± 6^a	175 ± 6^a	155 ± 6^a	209 ± 9^a	176 ± 10^a
组别	例数	剪切PARP			XIAP
组别	例数	12 h	24 h	48 h	12 h	24 h	48 h
假手术组	30	100 ± 7^a	101 ± 6^a	100 ± 7^a	100 ± 4^a	101 ± 4^a	102 ± 3^a
颅脑损伤组	30	278 ± 23	401 ± 32	354 ± 38	65 ± 5	43 ± 7	58 ± 5
UCF-101干预组	30	163 ± 7^a	218 ± 12^a	176 ± 12^a	74 ± 3^a	58 ± 6^a	70 ± 4^a

组别	例数	Omi/HtrA2			pro-caspase-3			pro-caspase-9
组别	例数	12 h	24 h	48 h	12 h	24 h	48 h	12 h	24 h	48 h
假手术组	30	100.0 ± 1.6^a	101.1 ± 1.5^a	101.5 ± 1.8^a	100 ± 6^a	100 ± 5^a	109 ± 5^a	100 ± 5^a	105 ± 4^a	107 ± 5^a
颅脑损伤组	30	165.6 ± 8.0	218.8 ± 8.8	204.0 ± 8.1	177 ± 4	239 ± 10	203 ± 8	182 ± 10	267 ± 13	204 ± 9
UCF-101干预组	30	129.9 ± 4.7^a	170.3 ± 6.8^a	151.7 ± 8.4^a	157 ± 5^a	193 ± 6^a	175 ± 6^a	155 ± 6^a	209 ± 9^a	176 ± 10^a
组别	例数	剪切PARP			XIAP
组别	例数	12 h	24 h	48 h	12 h	24 h	48 h
假手术组	30	100 ± 7^a	101 ± 6^a	100 ± 7^a	100 ± 4^a	101 ± 4^a	102 ± 3^a
颅脑损伤组	30	278 ± 23	401 ± 32	354 ± 38	65 ± 5	43 ± 7	58 ± 5
UCF-101干预组	30	163 ± 7^a	218 ± 12^a	176 ± 12^a	74 ± 3^a	58 ± 6^a	70 ± 4^a

组别	例数	caspase-3			caspase-9
组别	例数	12 h	24 h	48 h	12 h	24 h	48 h
假手术组	30	100 ± 6^a	101 ± 4^a	108 ± 5^a	100 ± 8^a	97 ± 7^a	99 ± 8^a
颅脑损伤组	30	179 ± 8	311 ± 19	215 ± 12	309 ± 26	594 ± 48	446 ± 48
UCF-101干预组	30	149 ± 4^a	228 ± 20^a	173 ± 6^a	210 ± 20^a	403 ± 33^a	289 ± 341^a

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