驱动蛋白家族成员5A介导的溶酶体功能在镉神经毒性中的作用

邓平; 陈梦妍; 谢佳; 田丽; 余争平; 皮会丰

doi:10.16016/j.2097-0927.202108210

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驱动蛋白家族成员5A介导的溶酶体功能在镉神经毒性中的作用

基础医学 | 更新时间：2026-01-09

- 驱动蛋白家族成员5A介导的溶酶体功能在镉神经毒性中的作用
- Effect of KIF5A mediated lysosomal functions in cadmium exposure-induced neurotoxicity in vitro
- 陆军军医大学学报 2022年44卷第4期页码：320-328
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)军事预防医学系军队劳动卫生学教研室，电磁辐射医学防护教育部重点实验室
- 作者简介：
  
  [皮会丰，E-mail: pihuifeng2010@163.com PI Huifeng, E-mail: pihuifeng2010@163.com
- 基金信息：
  
  国家自然科学基金面上项目(82173473);国家自然科学基金面上项目(81872596)
- DOI：10.16016/j.2097-0927.202108210
  中图分类号： R341;R992;R995
- 收稿：2021-08-30，
  
  修回：2021-11-12，
  
  纸质出版：2022-02-28
- 稿件说明：
移动端阅览
邓平, 陈梦妍, 谢佳, 等. 驱动蛋白家族成员5A介导的溶酶体功能在镉神经毒性中的作用[J]. 陆军军医大学学报, 2022,44(4):320-328.

Ping DENG, Menyan CHEN, Jia XIE, et al. Effect of KIF5A mediated lysosomal functions in cadmium exposure-induced neurotoxicity in vitro[J]. Journal of Army Medical University, 2022, 44(4): 320-328.
邓平, 陈梦妍, 谢佳, 等. 驱动蛋白家族成员5A介导的溶酶体功能在镉神经毒性中的作用[J]. 陆军军医大学学报, 2022,44(4):320-328. DOI： 10.16016/j.2097-0927.202108210.

Ping DENG, Menyan CHEN, Jia XIE, et al. Effect of KIF5A mediated lysosomal functions in cadmium exposure-induced neurotoxicity in vitro[J]. Journal of Army Medical University, 2022, 44(4): 320-328. DOI： 10.16016/j.2097-0927.202108210.

摘要

目的

探讨驱动蛋白家族成员5A(kinesin family member 5A，KIF5A)介导的溶酶体功能损伤在镉神经毒性中的作用。

方法

以原代培养的SPF级C57BL/6J小鼠皮层神经元为模型，分为对照组和镉暴露组。检测氯化镉暴露24 h后皮层神经元的半数抑制浓度(IC

)。进一步用1、2、3 μmol/L氯化镉处理神经元24 h，检测细胞存活率、神经元分化和溶酶体功能。RT-qPCR和Western blot检测KIF5A mRNA和蛋白的表达。腺病毒过表达KIF5A检测镉暴露后神经元的溶酶体功能、分化及细胞存活率。

结果

神经元暴露氯化镉24 h的半数抑制浓度IC

为2.9 μmol/L。与对照组相比，2、3 μmol/L镉暴露24 h后神经元细胞活力受到抑制(

＜0.01)，突起生长的总长度和分枝点数目显著减少(

＜0.01)。此外，与对照组相比，2、3 μmol/L氯化镉处理后显著抑制神经元溶酶体组织蛋白酶B(cathepsin B，CTSB)的活性和改变其酸性环境(

＜0.01)，但是对组织蛋白酶D(cathepsin D，CTSD)活性没有影响。镉暴露后KIF5A的mRNA及蛋白表达显著降低(

＜0.01)。过表达KIF5A可以显著改善镉暴露对神经元组织蛋白酶活性CTSB的抑制、稳定镉暴露造成的溶酶体pH值改变，拮抗镉暴露对神经元细胞活力及突起生长的抑制作用(

＜0.01)。

结论

镉暴露显著减少神经元中KIF5A蛋白的表达，从而损伤溶酶体功能，抑制神经元细胞活力及突起生长。

Abstract

Objective

To investigate the effect of kinesin family member 5A (KIF5A) mediated lysosomal functions in cadmium (Cd) exposure-induced neurotoxicity.

Methods

Primary cortical neurons isolated and primarily cultured from specific pa

thogen free (SPF) C57BL/6J mice were used and divided into control and Cd-exposed groups. The 50% inhibitory concentration (IC

) of cortical neurons after 24 h of Cd exposure was measured. Furthermore

the neurons were treated with 1

2 and 3 μmol/L cadmium chloride (CdCl

) for 24 h

and then cell viability

neuronal differentiation and lysosomal function were measured. RT-qPCR and Western blotting were used to detect the changes of KIF5A expression at mRNA and protein levels. The effects of overexpression of KIF5A by adenoviral vector on cell viability

neuronal differentiation and lysosomal function were also investigated in the Cd-exposed cortical neurons.

Results

The IC

value in neurons exposure to CdCl

for 24 h was determined to be 2.9 μmol/L. Compared with the control group

the cell viability was inhibited (

0.01) and the total length of protrusion growth and the number of branching points were obviously reduced (

0.01) after 2 and 3 μmol/L CdCl

exposure for 24 h. In addition

2 and 3 μmol/L CdCl

treatment also resulted in dramatically decreased lysosomal activity of cathepsin B (CTSB) and altered acidic environment when compared with the control group (

0.01). However

the activity cathepsin D(CTSD) was not significantly changed in CdCl

-exposed neuronal cells. More importantly

the mRNA and protein levels of KIF5A were remarkably decreased after Cd exposure (

0.01). Overexpression of KIF5A effectively increased the CTSB activity

stabilized the lysosomal pH value

and antagonized the inhibitory effect on cell viability and protrusion growth induced by Cd exposure (

0.01).

Conclusion

Cd exposure significantly reduces KIF5A protein expression in neurons

and thereby impairs lysosomal function and inhibits neuronal cell viability and protrusion growth.

关键词

Keywords

references

尚二萍, 许尔琪, 张红旗, 等. 中国粮食主产区耕地土壤重金属时空变化与污染源分析[J]. 环境科学, 2018, 39(10): 4670-4683. DOI: 10.13227/j.hjkx.201802139.

SHANG E P, XU E Q, ZHANG H Q, et al . Spatial-temporal trends and pollution source analysis for heavy metal contamination of cultivated soils in five major grain producing regions of China[J ] . Environ Sci, 2018, 39(10): 4670-4683. DOI: 10.13227/j.hjkx.201802139.

邓平, 陈梦妍, 谢佳, 等. 基于基因芯片分析急性镉暴露致大鼠肝损伤及其可能机制[J]. 第三军医大学学报, 2020, 42(16): 1641-1647. DOI: 10.16016/j.1000-5404.202004185.

DENG P, CHEN M Y, XIE J, et al . Microarray analysis of the mechanism of liver injury induced by acute cadmium exposure in rats[J ] . J Third Mil Med Univ, 2020, 42(16): 1641-1647. D OI: 10.16016/j.1000-5404.202004185.

LIU H, SU L, CHEN X, et al . Higher blood cadmium level is associated with greater cognitive decline in rural Chinese adults aged 65 or older[J ] . Sci Total Environ, 2021, 756: 144072. DOI: 10.1016/j.scitotenv.2020.144072.

PENG Y, LI Z, YANG X, et al . Relation between cadmium body burden and cognitive function in older men: a cross-sectional study in China[J ] . Chemosphere, 2020, 250: 126535. DOI: 10.1016/j.chemosphere.2020.126535.

WANG H, ZHANG L, ABEL G M, et al . Cadmium exposure impairs cognition and olfactory memory in male C57BL/6 mice[J ] . Toxicol Sci, 2018, 161(1): 87-102. DOI: 10.1093/toxsci/kfx202.

BALLABIO A, BONIFACINO J S. Lysosomes as dynamic regulators of cell and organismal homeostasis[J]. Nat Rev Mol Cell Biol, 2020, 21(2): 101-118. DOI: 10.1038/s41580-019-0185-4.

YANG Y, KLIONSKY D J. Autophagy and disease: unanswered questions[J]. Cell Death Differ, 2020, 27(3): 858-871. DOI: 10.1038/s41418-019-0480-9.

PI H, LI M, XIE J, et al . Transcription factor E3 protects again st cadmium-induced apoptosis by maintaining the lysosomal-mitochondrial axis but not autophagic flux in Neuro-2a cells[J ] . Toxicol Lett, 2018, 295: 335-350. DOI: 10.1016/j.toxlet.2018.07.015.

LI M, PI H, YANG Z, et al . Melatonin antagonizes cadmium-induced neurotoxicity by activating the transcription factor EB-dependent autophagy-lysosome machinery in mouse neuroblastoma cells[J ] . J Pineal Res, 2016, 61(3): 353-369. DOI: 10.1111/jpi.12353.

DE PACE R, BRITT D J, MERCURIO J, et al . Synaptic vesicle precursors and lysosomes are transportedby different mechanisms in the axon of mammalian neurons[J ] . Cell Rep, 2020, 31(11): 107775. DOI: 10.1016/j.celrep.2020.107775.

LIU M, PI H, XI Y, et al . KIF 5 A-dependent axonal transport deficiency disrupts autophagic flux in trimethyltin chloride-induced neurotoxicity[J ] . Autophagy, 2021, 17(4): 903-924. DOI: 10.1080/15548627.2020.1739444..

ZHOU C, LIU M, MEI X, et al . Histone hypoacetylation contributes to neurotoxicity induced by chronic nickel exposure in vivo and in vitro [J ] . Sci Total Environ, 2021, 783: 147014. DOI: 10.1016/j.scitotenv.2021.147014..

WANG Q W, WANG Y, WANG T, et al . Cadmium-induced autophagy promotes survival of rat cerebral cortical neurons by activating class Ⅲ phosphoinositide 3-kinase/beclin-1/B-cell lymphoma 2 signaling pathways[J ] . Mol Med Rep, 2015, 12(2): 2912-2918. DOI: 10.3892/mmr.2015.3755.

TANG J, DUAN W, DENG P, et al . Cadmium disrupts mitochondrial distribution and activates excessive mitochondrial fission by elevating cytosolic calcium independent of MCU-mediated mitochondrial calcium uptake in its neurotoxicity[J ] . Toxicology, 2021, 453: 152726. DOI: 10.1016/j.tox.2021.152726.

NORDMANN M, CABRERA M, PERZ A, et al . The Mon1-Ccz1 complex is the GEF of the late endosomal Rab7 homolog Ypt7[J ] . Curr Biol, 2010, 20(18): 1654-1659. DOI: 10.1016/j.cub.2010.08.002.

智翠娜, 来丽叶, 窦长松, 等. 溶酶体在锰致SK-N-SH细胞毒性中的作用[J]. 中华劳动卫生职业病杂志, 2019, 37(5): 332-336. DOI: 10.3760/cma.j.issn.1001-9391.2019.05.003.

ZHI C N, LAI L L, DOU C S, et al . The role of lysosomes in manganese-induced toxicity in SK-N-SH cells[J ] . Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi, 2019, 37(5): 332-336. DOI: 10.3760/cma.j.issn.1001-9391.2019.05.003.

GU X, HAN M, DU Y, et al . Pb disrupts autophagic flux through inhibiting the formation and activity of lysosomes in neural cells[J ] . Toxicol in Vitro , 2019, 55: 43-50. DOI: 10.10 16/j.tiv.2018.11.010..

WINDING M, KELLIHER M T, LU W, et al . Role of kinesin-1-based microtubule sliding in Drosophila nervous system development[J ] . Proc Natl Acad Sci U S A, 2016, 113(34): E4985-E4994. DOI: 10.1073/pnas.1522416113.

FERENZ N P, GABLE A, WADSWORTH P. Mitotic functions of kinesin-5[J]. Semin Cell Dev Biol, 2010, 21(3): 255-259. DOI: 10.1016/j.semcdb.2010.01.019.

XIA C H, ROBERTS E A, HER L S, et al . Abnormal neuro- filament transport caused by targeted disruption of neuronal kinesin heavy chain KIF 5 A[J ] . J Cell Biol, 2003, 161(1): 55-66. DOI: 10.1083/jcb.200301026..

WANG Q, TIAN J, CHEN H, et al . Amyloid beta-mediated KIF 5 A deficiency disrupts anterograde axonal mitochondrial movement[J ] . Neurobiol Dis, 2019, 127: 410-418. DOI: 10.1016/j.nbd.2019.03.021..

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