Protective effect of Humanin on rotenone-induced dopamine neuron toxicity

Yaohui SHAN; Qifu ZHANG; Jin CHENG; Feng YE; Xi ZHANG; Wenpei YU; Xiaogang WANG; Yuanpeng ZHAO; Guorong DAN; Mingliang CHEN; Yan SAI

doi:10.16016/j.2097-0927.202309083

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Protective effect of Humanin on rotenone-induced dopamine neuron toxicity

Neuroscience | 更新时间：2026-01-09

- Protective effect of Humanin on rotenone-induced dopamine neuron toxicity
- Journal of Army Medical University Vol. 46, Issue 7, Pages: 670-677(2024)
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)军事预防医学系防化医学教研室
- 作者简介：
- 基金信息：
  
  Supported the General Program of National Natural Science Foundation of China(81973090)
- DOI：10.16016/j.2097-0927.202309083
  CLC： R394.2;R742.5;R977.6
- Received：15 September 2023，
  
  Revised：2023-12-21，
  
  Published：15 April 2024
- 稿件说明：
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Yaohui SHAN, Qifu ZHANG, Jin CHENG, et al. Protective effect of Humanin on rotenone-induced dopamine neuron toxicity[J]. Journal of Army Medical University, 2024, 46(7): 670-677.
DOI：

Yaohui SHAN, Qifu ZHANG, Jin CHENG, et al. Protective effect of Humanin on rotenone-induced dopamine neuron toxicity[J]. Journal of Army Medical University, 2024, 46(7): 670-677. DOI： 10.16016/j.2097-0927.202309083.

摘要

目的

探讨Humanin(HN)对鱼藤酮(rotenone，Rot)诱导的多巴胺神经元毒性损伤的保护作用和机制。

方法

构建Rot染毒PC12细胞模型，实验设对照组、Rot染毒组、HN预处理Rot染毒组、单独HN处理组。采用ELISA检测Rot染毒细胞内外HN的含量，CCK-8法检测细胞活性，ATP检测试剂盒检测细胞内ATP含量，DCFH-DA探针检测细胞内ROS水平，Western blot分别检测线粒体自噬调控蛋白Pink1、Parkin、p-Parkin、p62、LC3，线粒体生物发生调控蛋白PGC1α和分裂/融合调控蛋白OPA1、MFN2、DRP1、p-DRP1以及抗氧化应激调控蛋白Keap1、Nrf2的表达。采用HBAD-mcherry-EGFP-LC3腺病毒转染细胞观察自噬体和自噬溶酶体的数量。

结果

Rot染毒组PC12细胞内HN浓度显著高于对照组(

0.05)；与对照组比较，Rot染毒组PC12细胞活性下降，ATP含量下降，ROS的生成增加，Rot染毒后PC12细胞内Pink1、p-Parkin表达升高，LC3Ⅱ/LC3Ⅰ比值升高，p62表达下降，细胞线粒体生物发生蛋白PGC1α和线粒体融合蛋白MFN2、OPA1表达下降，而线粒体分裂蛋白p-DRP1表达升高

抗氧化应激蛋白Keap1和Nrf2表达下降(

均

0.05)；与对照组比较，Rot染毒组PC12细胞中自噬体和自噬溶酶体数量增多(

0.05)，而20 μmol/L HN预处理可以改善Rot染毒引起的上述变化(

0.05)。

结论

HN通过抑制线粒体自噬和线粒体分裂、促进线粒体生物发生和融合以及抗氧化应激，改善Rot诱导的多巴胺神经元毒性损伤。

Abstract

Objective

To investigate the mechanism and protective effect of Humanin (HN) on rotenone (Rot)-induced toxic damage for dopamine neurons.

Methods

The Rot-poisened PC12 cell model was constructed

and the control group

the Rot poisening group

the HN pretreated Rot poisening group

and the HN treatment group were set up. ELISA was used to detect the content of HN inside and outside of Rot-infected cells

CCK-8 assay was used to detect cell viability

and ATP detection kit was used to detect the intracellular ATP content. Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay was used to detect the level of reactive oxygen species (ROS) in cells. Western blotting was performed to detect the expression level of mitochondrial autophagy regulatory proteins Pink1

Parkin

p62

LC3

mitochondrial biogenesis regulatory protein PGC1α

division/fusion regulatory proteins OPA1

MFN2

DRP1

p-DRP1 and antioxidant stress regulatory proteins Keap1 and Nrf2. HBAD-mcherry-EGFP-LC3 adenovirus transfected cells was used to observed the number of autophagosomes and autophagolysosomes.

Results

The results showed that the intracellular concentration of HN in PC12 in the Rot poisening group was significantly higher than that in the control group (

0.05);Compared with the control group

the Rot poisening group had significantly decreased activity of PC12 cells

decreased ATP content and increased production of ROS. After the poisen of Rot in PC12 cells

the expression of Pink1 and p-Parkin

the ratio of LC3Ⅱ/LC3Ⅰ and the expression of p-DRP1 in mitochondrial fusion protein was increased

while the expression of p62

the expression of mitochondrial biogenesis protein PGC1α

mitochondrial fusion proteins MFN2 and OPA1

and antioxidant stress proteins Keap1 and Nrf2 were decreased (al

0.05). The number of autophagosomes and autophagolysosomes in PC12 cells in the Rot poisening group was higher than that in the control group (

0.05)

and HN pretreatment (20 μmol/L) could significantly improve the changes mentioned above caused by Rot poisening (

0.05).

Conclusion

HN ameliorates Rot-induced toxic damage for dopamine neurons by inhibiting mitophagy and mitochondrial division and promoting mitochondrial biogenesis and fusion

and anti-oxidative stress.

关键词

Keywords

references

BISBAL M, SANCHEZ M. Neurotoxicity of the pesticide rotenone on neuronal polarization: a mechanistic approach[J]. Neural Regen Res, 2019, 14(5): 762-766. DOI: 10.4103/1673-5374.249847.

RADAD K, AL-SHRAIM M, AL-EMAM A, et al . Rotenone: from modelling to implication in Parkinson's disease[J ] . Folia Neuropathol, 2019, 57(4): 317-326. DOI: 10.5114/fn.2019.89857.

OLUBODUN-OBADUN T G, ISHOLA I O, ADEYEMI O O. Impact of environmental toxicants exposure on gut-brain axis in Parkinson disease[J]. Drug Metab Pers Ther, 2022, 37(4): 329-336. DOI: 10.1515/dmpt-2021-0144.

PENG K G, XIAO J S, YANG L K, et al . Mutual antagonism of PINK1/parkin and PGC-1α contributes to maintenance of mitochondrial homeostasis in rotenone-induced neurotoxicity[J ] . Neurotox Res, 2019, 35(2): 331-343. DOI: 10.1007/s12640-018-9957-4.

SHARMA S, PATEL F, ARA H, et al . Rotenone-induced 4-HNE aggresome formation and degradation in HL-1 cardiomyocytes: role of autophagy flux[J ] . Int J Mol Sci, 2022, 23(9): 4675. DOI: 10.3390/ijms23094675.

CORADDUZZA D, CONGIARGIU A, CHEN Z C, et al . Humanin and its pathophysiological roles in aging: a systematic review[J ] . Biology (Basel), 2023, 12(4): 558. DOI: 10.3390/biology12040558.

ROCHETTE L, MELOUX A, ZELLER M, et al . Role of humanin, a mitochondrial-derived peptide, in cardiovascular disorders[J ] . Arch Cardiovasc Dis, 2020, 113(8/9): 564-571. DOI: 10.1016/j.acvd.2020.03.020.

THIANKHAW K, CHATTIPAKORN K, CHATTIPAKORN S C, et al . Roles of humanin and derivatives on the pathology of neurodegenerative diseases and cognition[J ] . Biochim Biophys Acta Gen Subj, 2022, 1866(4): 130097. DOI: 10.1016/j.bbage n.2022.130097.

ZAPAŁA B, KACZYNSKI Ł, KIEC-WILK B, et al . Humanins, the neuroprotective and cytoprotective peptides with antiapoptotic and anti-inflammatory properties[J ] . Pharmacol Rep, 2010, 62(5): 767-777. DOI: 10.1016/s1734-1140(10)70337-6.

WANG Y Y, LI N Y, ZENG Z Y, et al . Humanin regulates oxidative stress in the ovaries of polycystic ovary syndrome patients via the Keap1/Nrf2 pathway[J ] . Mol Hum Reprod, 2021, 27(2): gaaa081. DOI: 10.1093/molehr/gaaa081.

WANG T, ZHANG L, ZHANG M, et al . Gly14 ] -Humanin reduces histopathology and improves functional outcome after traumatic brain injury in mice[J ] . Neuroscience, 2013, 231: 70-81. DOI: 10.1016/j.neuroscience.2012.11.019.

YANG X R, ZHANG H M, WU J Z, et al . Humanin attenuates NMDA-induced excitotoxicity by inhibiting ROS-dependent JNK/p38 MAPK pathway[J ] . Int J Mol Sci, 2018, 19(10): 2982. DOI: 10.3390/ijms19102982.

XU X S, CHUA C C, GAO J P, et al . Neuroprotective effect of humanin on cerebral ischemia/reperfusion injury is mediated by a PI3K/Akt pathway[J ] . Brain Res, 2008, 1227: 12-18. DOI: 10.1016/j.brainres.2008.06.018.

SIMON D K, TANNER C M, BRUNDIN P. Parkinson disease epidemiology, pathology, genetics, and pathophysiology[J]. Clin Geriatr Med, 2020, 36(1): 1-12. DOI: 10.1016/j.cger.2019.08.002.

MALPARTIDA A B, WILLIAMSON M, NARENDRA D P, et al . Mitochondrial dysfunction and mitophagy in parkinson's disease: from mechanism to therapy[J ] . Trends Biochem Sci, 2021, 46(4): 329-343. DOI: 10.1016/j.tibs.2020.11.007.

DORSZEWSKA J, KOWALSKA M, PRENDECKI M, et al . Oxidative stress factors in Parkinson's disease[J ] . Neural Regen Res, 2021, 16(7): 1383. DOI: 10.4103/1673-5374.300980.

ELDEEB M A, THOMAS R A, RAGHEB M A, et al . Mitochondrial quality control in health and in Parkinson's disease[J ] . Physiol Rev, 2022, 102(4): 1721-1755. DOI: 10.1152/physrev.00041.2021.

QIN Q Q, JIN J Q, HE F, et al . Humanin promotes mitochondrial biogenesis in pancreatic MIN 6 β-cells[J ] . Biochem Biophys Res Commun, 2018, 497(1): 292-297. DOI: 10.1016/j.bbrc.2018.02.071..

HSU H T, YANG Y L, CHANG W H, et al . Hyperbaric oxygen therapy improves Parkinson's disease by promoting mitochondrial biogenesis via the SIRT-1/PGC-1α pathway[J ] . Biomolecules, 2022, 12(5): 661. DOI: 10.3390/biom12050661..

ZHANG X L, HUANG W M, SHAO Q H, et al . Drp1, a potential therapeutic target for Parkinson's disease, is involved in olfactory bulb pathological alteration in the Rotenone-induced rat model[J ] . Toxicol Lett, 2020, 325: 1-13. DOI: 10.1016/j.toxlet.2020.02.009.

LI J, YANG D M, LI Z P, et al . PINK1/parkin-mediated mitophagy in neurodegenerative diseases[J ] . Ageing Res Rev, 2023, 84: 101817. DOI: 10.1016/j.arr.2022.101817.

RAKOVIC A, ZIEGLER J, MÅRTENSSON C U, et al . PINK1-dependent mitophagy is driven by the UPS and can occur independently of LC3 conversion[J ] . Cell Death Differ, 2019, 26(8): 1428-1441. DOI: 10.1038/s41418-018-0219-z.

ZHOU Q, ZHANG Y, LU L, et al . Copper induces microglia-mediated neuroinflammation through ROS/NF-κB pathway and mitophagy disorder[J ] . Food Chem Toxicol, 2022, 168: 113369. DOI: 10.1016/j.fct.2022.113369.

FAN H J, LI Y R, SUN M Y, et al . Hyperoside reduces rotenone-induced neuronal injury by suppressing autophagy[J ] . Neurochem Res, 2021, 46(12): 3149-3158. DOI: 10.1007/s11064-021-03404-z.

ARAB H H, SAFAR M M, SHAHIN N N. Targeting ROS-dependent AKT/GSK-3β/NF-κB and DJ-1/Nrf2 pathways by dapagliflozin attenuates neuronal injury and motor dysfunction in rotenone-induced Parkinson's disease rat model[J]. ACS Chem Neurosci, 2021, 12(4): 689-703. DOI: 10.1021/acschemneuro.0c00722.

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