吉林大学学报(医学版) ›› 2022, Vol. 48 ›› Issue (3): 728-733.doi: 10.13481/j.1671-587X.20220322

• 基础研究 • 上一篇    

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

潘琪1,王子珍1,叶富跃1,邢伟1,林家汉1,卢剪月2,杨堃1()   

  1. 1.海南医学院第一附属医院神经外科,海南 海口 570102
    2.海南医学院第一附属医院康复医学科,海南 海口 570102
  • 收稿日期:2021-11-14 出版日期:2022-05-28 发布日期:2022-06-21
  • 通讯作者: 杨堃 E-mail:chbyk1379@163.com
  • 作者简介:潘 琪(1981-),男,安徽省桐城市人,副主任医师,医学博士,主要从事功能神经外科疾病方面的研究。
  • 基金资助:
    海南省卫健委卫生计生行业科研项目(20A200319);海南省教育厅高等学校科学研究项目(Hnky-2021-38);海南省科技厅自然科学基金高层次人才项目(821RC698);海南医学院科研培育基金项目(HY2015-19)

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

Qi PAN1,Zizhen WANG1,Fuyue YE1,Wei XING1,Jiahan LIN1,Jianyue LU2,Kun YANG1()   

  1. 1.Department of Neurosurgery,First Affiliated Hospital,Hainan Medical University,Haikou 570102,China
    2.Department of Rehabilitation Medicine,First Affiliated Hospital,Hainan Medical University,Haikou 570102,China
  • Received:2021-11-14 Online:2022-05-28 Published:2022-06-21
  • Contact: Kun YANG E-mail:chbyk1379@163.com

摘要: 目的

观察蛋白酶体抑制剂乳胞素对大鼠黑质多巴胺能神经元死亡和氧化损伤的作用,并探讨氧化损伤在蛋白酶体抑制剂诱导多巴胺能神经元死亡中的作用。

方法

将70只SD大鼠随机分为乳胞素组和生理盐水组,每组35只。将蛋白酶体抑制剂乳胞素立体定向注射入乳胞素组大鼠右侧黑质中,生理盐水组大鼠注射等体积的生理盐水。旷场实验检测2组大鼠的穿梭距离、下肢站立次数和总跑动时间,前肢使用不对称实验检测2组大鼠前肢使用不对称指数,阿扑吗啡诱导旋转实验检测2组大鼠旋转行为,微量酶标法检测2组大鼠中脑黑质中羟自由基、还原型谷胱甘肽(GSH)、丙二醛(MDA)、蛋白质羰基和8-羟基脱氧鸟苷(8-OHdG)水平,免疫组织化学染色检测2组大鼠黑质中酪氨酸羟化酶(TH)免疫阳性神经元毁损程度。

结果

与生理盐水组比较,乳胞素组大鼠的穿梭距离、下肢站立次数和总跑动时间明显减少(P<0.05)。与生理盐水组比较,乳胞素组大鼠前肢使用不对称指数明显升高(P<0.05)。乳胞素组大鼠注射阿扑吗啡后出现恒向健侧旋转。给药7和21 d后,与生理盐水组比较,乳胞素组大鼠注射侧黑质中羟自由基、MDA、蛋白质羰基和8-OHdG水平明显升高(P<0.05),GSH水平明显降低(P<0.05)。给药21 d后,乳胞素组大鼠注射侧黑质中TH免疫阳性神经元显著丧失。与生理盐水组比较,乳胞素组大鼠黑质中TH免疫阳性神经元残存率明显降低(P<0.05)。

结论

大鼠黑质内注射蛋白酶体抑制剂乳胞素能够诱导大鼠黑质多巴胺能神经元氧化损伤,并最终导致神经元死亡。

关键词: 帕金森病, 蛋白酶体抑制剂, 氧化损伤, 多巴胺能神经元, 乳胞素

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

中图分类号: 

  • R742.5