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Enhancing the Astrocytic Clearance of Extracellular α-Synuclein Aggregates by Ginkgolides Attenuates Neural Cell Injury

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Abstract

The accumulation of aggregated forms of the α-Synuclein (α-Syn) is associated with the pathogenesis of Parkinson’s disease (PD), and the efficient clearance of aggregated α-Syn represents a potential approach in PD therapy. Astrocytes are the most numerous glia cells in the brain and play an essential role in supporting brain functions in PD state. In the present study, we demonstrated that cultured primary astrocytes engulfed and degraded extracellular aggregated recombinant human α-Syn. Meanwhile, we observed that the clearance of α-Syn by astrocytes was abolished by proteasome inhibitor MG132 and autophagy inhibitor 3-methyladenine (3MA). We further showed that intracellular α-Syn was reduced after ginkgolide B (GB) and bilobalide (BB) treatment, and the decrease was reversed by MG132 and 3MA. More importantly, GB and BB reduced indirect neurotoxicity to neurons induced by α-Syn-stimulated astrocytic conditioned medium. Together, we firstly find that astrocytes can engulf and degrade α-Syn aggregates via the proteasome and autophagy pathways, and further show that GB and BB enhance astrocytic clearance of α-Syn, which gives us an insight into the novel therapy for PD in future.

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Abbreviations

α-Syn:

α-Synuclein

ALP:

Autophagy–lysosome pathways

BB:

Bilobalide

BCA:

Bicinchoninic acid

CM:

Conditioned medium

DAPI:

4,6-Linked amidine-2-phenylindole

DLB:

Dementia with LBs

DMSO:

Dimethyl sulfoxide

DTT:

DL-Dithiothreitol

GB:

Ginkgolide B

GBE:

Ginkgo biloba extract

GFAP:

Glial fibrillary acidic protein

LBs:

Lewy bodies

LC3:

Microtubule-associated protein 1A/1B-light chain 3

LDH:

Lactate dehydrogenase

LNs:

Lewy neuritis

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium

PBS:

Phosphate buffered saline

PD:

Parkinson’s disease

SNpc:

Substantia Nigra pars compacta

ThT:

Thioflavin T

3MA:

3-Methyladenine

UPS:

Ubiquitin proteasome system

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Acknowledgements

We are thankful to Prof. Zhuohua Zhang for providing the plasmid pcDNA3.1(−)-α-Syn. This research was supported by the grants from the National Natural Science Foundation of China (Grant Nos. 81673408 and 81703485).

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Author notes

  1. Jun Hua and Nuo Yin have contributed equally to this work.

Authors and Affiliations

  1. Jiangsu Key Laboratory of Neurodegeneration, Department of Pharmacology, Nanjing Medical University, 101 Longmian Road, Nanjing, 211166, Jiangsu, China

    Jun Hua, Nuo Yin, Shi Xu, Qiang Chen, Tingting Tao, Jianhua Ding, Yi Fan & Gang Hu

  2. Department of Neurology & Psychology, The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, 1 Fuhua Road, Shenzhen, 518033, Guangdong, China

    Jun Hua

  3. Neuroprotective Drug Discovery Key Laboratory of Nanjing Medical University, 101 Longmian Road, Nanjing, 211166, Jiangsu, China

    Qiang Chen, Tingting Tao & Yi Fan

  4. Division of Clinical Pharmacy, Department of Pharmacy, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, Jiangsu, China

    Ji Zhang

  5. Nanjing University of Chinese Medicine, 138 Xianlin Avenue, Nanjing, 210023, Jiangsu, China

    Gang Hu

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  1. Jun Hua
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Contributions

YF and GH conceived the study and analyzed the data. JH and NY constructed lentiviral vectors and involved in manuscript preparation. JH and SX designed and performed animal experiment. NY and QC performed biochemical analysis. TT and JZ performed immunohistochemical and immunolabeling analysis. JH and JD performed in vitro cell culture experiments. NY and JD performed in vitro α-synuclein propagation assay. JH and YF wrote the manuscript. All authors reviewed and approved the final manuscript.

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Correspondence to Yi Fan or Gang Hu.

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The authors declare that they have no conflict of interests.

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All study protocols were approved by the Institutional Animal Care and Use Committee of Nanjing Medical University (IACUC).

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Hua, J., Yin, N., Xu, S. et al. Enhancing the Astrocytic Clearance of Extracellular α-Synuclein Aggregates by Ginkgolides Attenuates Neural Cell Injury. Cell Mol Neurobiol 39, 1017–1028 (2019). https://doi.org/10.1007/s10571-019-00696-2

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