蛋白酶体抑制剂乳胞素对大鼠黑质多巴胺能神经元氧化损伤的作用

doi:10.13481/j.1671-587X.20220322

Abstract

Abstract: Objective

To observe the effect of proteasome inhibitor lactacystin on the death and oxidative damage of dopaminergic neurons in substantia nigra of the rats， and to explore the effect of oxidative damage on the death of dopaminergic neurons induced by proteasome inhibitor.

Methods

Seventy SD rats were randomly divided into lactacystin group and normal saline group，and there were 35 rats in each group. The proteasome inhibitor lactacystin was injected into the right substantia nigra of the rats in lactacystin group， and the rats in control group were injected with the same volume of normal saline. The shuttle distance， the number of lower limb standing， and the total running time of the rats in two groups were detected by open field experiment； the forelimb asymmetry indexes of the rats in two groups were detected by forelimb asymmetry experiment； apomorphine induced rotation experiment was used to test the rotation behavior of the rats in two groups. The levels of hydroxyl free radical， reduced glutathione（GSH）， malondialdehyde（MDA）， protein carbonyl and 8-hydroxydeoxyguanosine（8-OHdG） in substantia nigra of the rats in two groups were detected by microplate method； the damage of tyrosine hydroxylase（TH） immunopositive neurons in substantia nigra of the rats in various groups was detected by immunohistochemistry.

Results

Compared with normal saline group， the shuttle distance， the number of standing， and the total running time of the rats in lactacystin group were significantly decreased（P<0.05）. The forelimb asymmetry index of the rats in lactacystin group was significantly higher than that in normal saline group （P<0.05）.After injected with apomorphine， the rats in lactacystin group showed constant rotation to the healthy side.After administrated for 7 and 21 d， compared with normal saline group， the levels of hydroxyl radical， MDA， protein carbonyl， and 8-OHdG in right substantia nigra of the rats in lactacystin group were increased significantly （P<0.05），while the levels of GSH were decreased significantly （P<0.05）.After administrated for 21 d， the TH immunopositive neurons in right substantia nigra of the rats in lactacystin group were significantly lost. Compared with normal saline group， the survival rate of TH immunopositive neurons in substantia nigra of the rats in lactacystin group was significantly decreased （P<0.05）.

Conclusion

Injection of proteasome inhibitor lactosin can induce the oxidative damage of dopaminergic neurons in substantia nigra of the rats， and eventually lead to the death of neurons.

Key words: Parkinson’s disease, Proteasome inhibitor, Oxidative damage, Dopaminergic neuron, Lactacystin

CLC Number:

R742.5

Qi PAN,Zizhen WANG,Fuyue YE,Wei XING,Jiahan LIN,Jianyue LU,Kun YANG. Effect of proteasome inhibitor lactacystin on oxidative damage of dopaminergic neurons in substantia nigra of rats[J].Journal of Jilin University(Medicine Edition), 2022, 48(3): 728-733.

Figures/Tables 4

Tab. 1

Tab. 2

Tab. 3

Fig. 1

References 21

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Group	Shuttle distance (Number of units)	Number of standing	Total running time (t/s)
Normal saline	277.10±30.05	44.50±11.62	416.50±56.13
Lactacystin	80.50±14.86^*	17.40±6.48^*	183.80±30.32^*
^P*<0.05 compared with normal saline group.

Group	Hydroxyl radical [λ_B/(U·mg^－1)]			GSH [m_B/(μmol·g^－1)]
Group	(t/d) 3	7	21	(t/d) 3	7	21
Normal saline	13.67±2.87	14.48±3.87	13.16±3.72	1.26±0.37	1.35±0.40	1.30±0.33
Lactacystin	14.26±2.92	40.35±5.78 ^*	104.41±5.20 ^*	1.31±0.42	0.83±0.23 ^*	0.68±0.14 ^*
^P*<0.05 compared with normal saline group.

Group	MDA[m_B /(μmol·g^－1)]			Protein carbonyl [m_B/(μmol·g^－1)]			8-OHdG[w_B/(ng·g^－1)]
Group	(t/d) 3	7	21	(t/d) 3	7	21	(t/d) 3	7	21
Normal saline	6.16±2.20	6.00±2.18	5.74±2.02	8.53±1.29	7.88±1.37	9.20±1.33	20.29±3.81	17.91±3.57	22.31±3.46
Lactacystin	6.30±2.17	10.03±3.45^*	12.38±3.59^*	9.46±1.48	15.09±2.15^*	18.13±2.77^*	22.32±3.64	40.08±7.39^*	61.41±8.49^*
^P*<0.05 compared with normal saline group.

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Effect of proteasome inhibitor lactacystin on oxidative damage of dopaminergic neurons in substantia nigra of rats

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