Protective Effect and Mechanism of Velvet Antler Peptide on Rotenone-induced SH-SY5Y Cell Damage

Xu-fan ZHANG; Jun-ling CAO; Jin-tao HU; Wei LIU; Tian-quan WANG; Jing-feng OUYANG

doi:10.13422/j.cnki.syfjx.20201974

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Protective Effect and Mechanism of Velvet Antler Peptide on Rotenone-induced SH-SY5Y Cell Damage

Pharmacology | 更新时间：2020-12-10

- Protective Effect and Mechanism of Velvet Antler Peptide on Rotenone-induced SH-SY5Y Cell Damage
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 24, Pages: 82-88(2020)
- 作者机构：
  
  1.北京中医药大学东直门医院，北京 100700
  2.北京中医药大学东方医院，北京 100078
  3.中国中医科学院医学实验中心，北京 100700
  4.北京中医药大学中药学院，北京 102488
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20201974
  CLC： R22;R242;R2-031;R285.5
- Received：24 February 2020，
  
  Published Online：27 July 2020，
  
  Published：20 December 2020
- 稿件说明：
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张旭帆,曹俊岭,胡金涛等.鹿茸多肽对鱼藤酮致SH-SY5Y细胞损伤的保护作用及其机制[J].中国实验方剂学杂志,2020,26(24):82-88.

ZHANG Xu-fan,CAO Jun-ling,HU Jin-tao,et al.Protective Effect and Mechanism of Velvet Antler Peptide on Rotenone-induced SH-SY5Y Cell Damage[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(24):82-88.
张旭帆,曹俊岭,胡金涛等.鹿茸多肽对鱼藤酮致SH-SY5Y细胞损伤的保护作用及其机制[J].中国实验方剂学杂志,2020,26(24):82-88. DOI： 10.13422/j.cnki.syfjx.20201974.

ZHANG Xu-fan,CAO Jun-ling,HU Jin-tao,et al.Protective Effect and Mechanism of Velvet Antler Peptide on Rotenone-induced SH-SY5Y Cell Damage[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(24):82-88. DOI： 10.13422/j.cnki.syfjx.20201974.

摘要

目的

探讨鹿茸多肽对鱼藤酮诱导的神经母细胞瘤（SH-SY5Y）细胞损伤的干预作用及相关机制。

方法

用0.5 μmol·L

-1

的鱼藤酮作用于SH-SY5Y细胞建立帕金森病（PD）体外模型，设置空白组、模型组、鹿茸多肽高、中、低剂量（150，100，50 g·L

-1

）组及雷帕霉素组。通过苏木精-伊红（HE）染色，罗丹明123（Rh123）染色，2'，7'-二氯荧光黄二乙酸酯（DCFH-DA）染色及免疫组织化学染色观察各组细胞中路易小体的数量、线粒体膜电位的变化、活性氧簇（ROS）含量及

-突触核蛋白（

-syn），蛋白激酶B（Akt），哺乳动物雷帕霉素靶蛋白（mTOR）的表达情况。

结果

HE染色结果显示，与空白组比较，模型组中细胞数目减少、触角结构变钝，形态变圆，胞质内可见嗜酸性路易小体。与模型组比较，雷帕霉素组及鹿茸多肽高、中、低剂量组中细胞形态并无明显差异，但是胞质中路易小体较少；罗丹明123染色结果显示，与空白组比较，模型组中细胞线粒体膜电位明显升高（

0.05）。与模型组比较，雷帕霉素组及鹿茸多肽高、中、低剂量组中细胞线粒体膜电位均有所下降（

0.05）；DCFH-DA染色结果显示，与空白组比较，模型组细胞中活性氧（ROS）含量明显增多（

0.05）；与模型组比较，雷帕霉素组及鹿茸多肽高、中、低剂量组的活性氧（ROS）含量均下降（

0.05）。免疫组织化学结果显示，与空白组比较，模型组

-syn，Akt，mTOR蛋白表达明显增多（

0.05）；与模型组比较，雷帕霉素组及鹿茸多肽高、中剂量组

-syn，mTOR蛋白表达均明显减少（

0.05），雷帕霉素组及鹿茸多肽高剂量组中Akt表达明显减少（

0.05）。

结论

鹿茸多肽可能是通过抑制Akt/mTOR信号通路促进

-syn的降解，从而发挥神经元保护作用。

Abstract

Objective

To investigate the intervening effect of velvet antler peptide （VAP） on rotenone-induced neuroblastoma （SH-SY5Y） cell damage and explore its related mechanism.

Method

0.5 μmol·L

-1

rotenone was used to SH-SY5Y cells to establish an

in vitro

model of Parkinson's disease （PD）. A blank control group， a model group， high， medium and low dose VAP groups （150，100，50 mg·L

-1

， respectively） and a rapamycin group were established. The number of lewy bodies， changes in mitochondrial membrane potential， content of reactive oxygen species （ROS） and

α-

synuclein （

-syn）， protein kinase B （Akt）， and mammalian target of rapamycin （mTOR） were observed by hematoxylin-eosin（HE） staining， rhodamine 123 staining， DCFH-DA staining and immunohistochemical staining expression respectively.

Result

The results of HE staining showed that as compared with the blank group， the number of cells in model group was reduced， the tentacle structure became dull， the shape became round， and eosinophilic Lewy bodies were visible in cytoplasm. As compared with model group， there was no significant difference in cell morphology from rapamycin group and VAP high， medium and low dose groups， but there were fewer Lewy bodies in cytoplasm in these four groups. Rhodamine 123 staining showed that as compared with blank group， the mitochondrial membrane potential was increased significantly in model group （

0.05）. As compared with the model group， the mitochondrial membrane potential was decreased in rapamycin group and VAP high， medium and low dose groups （

0.05）. DCFH-DA staining results showed that as compared with blank group， the content of ROS was increased significantly in cells of model group （

0.05）. As compared with model group， the content of ROS was decreased in rapamycin group and VAP high， medium and low dose groups （

0.05）. Immunohistochemical staining showed that as compared with blank group， the protein expression levels of

-syn，Akt，and mTOR were increased significantly in model group （

0.05）. As compared with model group， the protein expression levels of

-syn and mTOR were significantly reduced in rapamycin group and VAP high and medium dose groups （

0.05）， and the expression levels of Akt were significantly reduced in rapamycin group and VAP high-dose group （

0.05）.

Conclusion

Velvet antler peptides may play a neuroprotective role by regulating the Akt/mTOR signaling pathway and promoting the degradation of

-syn in SH-SY5Y cells.

关键词

Keywords

references

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