中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2017, Vol. 12 ›› Issue (7): 1145-1151.doi: 10.4103/1673-5374.211195

• 原著:退行性病与再生 • 上一篇    下一篇

保守性多巴胺神经营养因子如何保护和拯救变性的PC12细胞？

  

  • 收稿日期:2017-05-06 出版日期:2017-07-15 发布日期:2017-07-15
  • 基金资助:

    安徽省自然科学基金（11040606Q11）; 国家自然科学基金（81100960）

How does conserved dopamine neurotrophic factor protect against and rescue neurodegeneration of PC12 cells?

Jia-ming Mei1, 2, Chao-shi Niu1, 2   

  1. 1 Shandong University, Jinan, Shandong Province, China; 2 Department of Neurosurgery, Anhui Provincial Hospital, Hefei, Anhui Province, China
  • Received:2017-05-06 Online:2017-07-15 Published:2017-07-15
  • Contact: Chao-shi Niu, Ph.D., doctorstar@126.com.
  • Supported by:

    This study was supported by the Natural Science Foundation of Anhui Province of China, No. 11040606Q11; the National Natural Science Foundation of China, No. 81100960.

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摘要：

保守性多巴胺神经营养因子已经在帕金森病动物模型中证实对多巴胺能神经元具有保护作用，但具体机制不明。我们的实验希望通过不可逆性蛋白酶体抑制剂Lactacystin和可逆性蛋白酶体抑制MG132分别诱导PC12细胞变性，采用保守性多巴胺神经营养因子蛋白分别进行预处理和后处理发现，（1）PC12细胞活性均提高、细胞形态改善，细胞中alpha-共核蛋白表达均减少；（2）ELISA分析显示，保守性多巴胺神经营养因子预处理和后处理均能上调MG132诱导的PC12细胞中26s蛋白酶体的表达，而在Lactacystin诱导的PC12细胞中，保守性多巴胺神经营养因子预处理可上调26s蛋白酶体表达；（3）对于3种蛋白水解酶(谷氨酰肽水解酶、糜蛋白酶和胰蛋白酶)而言，保守性多巴胺神经营养因子的上述两种干预方法均能上调谷氨酰肽水解酶的表达，而糜蛋白酶仅在保守性多巴胺神经营养因子预处理的MG132诱导PC12细胞中表达上调，胰蛋白酶表达无显著性变化；（4）结果证实，保守性多巴胺神经营养因子可通过调控蛋白酶体活性，保护和拯救PC12细胞的神经变性。 

ORCID:0000-0002-5429-1443(Chao-shi Niu)

关键词: 神经再生, 保守性多巴胺能神经营养因子, 帕金森病, 蛋白酶体抑制剂, 26 s蛋白酶, α-突触核蛋白, 动物模型, MG-132, 谷氨酰肽水解酶, 糜蛋白酶, 胰蛋白酶

Abstract:

Conserved dopamine neurotrophic factor protects and rescues dopaminergic neurodegeneration induced by 6-hydroxydopamine in vivo, but its potential value in treating Parkinson’s disease remains controversial. Here, we used the proteasome inhibitors lactacystin and MG132 to induce neurodegeneration of PC12 cells. Afterwards, conserved dopamine neurotrophic factor was administrated as a therapeutic factor, both pretreatment and posttreatment. Our results showed that (1) conserved dopamine neurotrophic factor enhanced lactacystin/MG132-induced cell viability and morphology, and attenuated alpha-synuclein accumulation in differentiated PC12 cells. (2) Enzyme linked immunosorbent assay showed up-regulated 26S proteasomal activity in MG132-induced PC12 cells after pre- and posttreatment with conserved dopamine neurotrophic factor. Similarly, 26S proteasome activity was upregulated in lactacystin-induced PC12 cells pretreated with conserved dopamine neurotrophic factor. (3) With regard proteolytic enzymes (specifically, glutamyl peptide hydrolase, chymotrypsin, and trypsin), glutamyl peptide hydrolase activity was up-regulated in lactacystin/MG132-administered PC12 cells after pre- and posttreatment with conserved dopamine neurotrophic factor. However, upregulation of chymotrypsin activity was only observed in MG132-administered PC12 cells pretreated with conserved dopamine neurotrophic factor. There was no change in trypsin expression. We conclude that conserved dopamine neurotrophic factor develops its neurotrophic effects by modulating proteasomal activities, and thereby protects and rescues PC12 cells against neurodegeneration.

Key words: nerve regeneration, conserved dopamine neurotrophic factor, Parkinson’s disease, proteasomal inhibitor, 26S proteasome, alpha- synuclein, lactacystin, MG-132, glutamyl peptide hydrolase, chymotrypsin, trypsin, neural regeneration