Ganoderma lucidum extract regulates autophagy induced by MPP+ in Neuro-2a cells

doi:10.3969/j.issn.1006-7795.2019.04.019

Abstract

Abstract: Objective To investigate the protective effect of Ganoderma lucidum extract (GLE) on autophagy of Neuro-2a cells induced by 1-methyl-4-phenylpyridinium (MPP⁺). Methods Western blotting was used to detect the conversion of autophagy marker LC3 (LC3-Ⅱ/LC3-I), and fluorescence microscopy was employed to detect the formation of LC3 punctate aggregates. Western blot was used to detect the expression levels of AMPK/mTOR/ULK1, PINK1/Parkin and NIX/LC3 proteins. Results The laser confocal microscopy showed that LC3-Ⅱ cells in the MPP⁺ treatment group were clustered on the autophagosome membrane, with a point-like aggregation state observed, while no obvious point aggregation was observed in the GLE intervention group. The level of autophagy was evaluated as the ratio of point-aggregated positive cells of LC3 over the total cells. According to statistics, the percentage of LC3 positive cells with green dot distribution in the MPP⁺ group was significantly increased. This value was significantly reduced in the GLE group. Western blotting results showed that the ratio of LC3-Ⅱ/LC3-I in GLE group was significantly decreased, and the expression level of NIX protein was significantly increased. After GLE treatment, the abnormal expression of each protein in AMPKα/mTOR/ULK1 and PINK1/Parkin pathway caused by MPP⁺ injury could be significantly improved.Conclusion GLE inhibited the autophagy response of MPP⁺-induced Neuro-2a cells, regulated the abnormal expression of autophagy regulatory proteins AMPK/mTOR/ULK1 and PINK1/Parkin, suggesting that GLE may play a neuroprotective role by regulating autophagy.

Key words: Ganoderma lucidum, Parkinson's disease, autophagy, neuroprotection

CLC Number:

R592

Ding Hui, Cai Yanning, Chen Biao. Ganoderma lucidum extract regulates autophagy induced by MPP⁺ in Neuro-2a cells[J]. Journal of Capital Medical University, 2019, 40(4): 596-601.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2019.04.019

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2019/V40/I4/596

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Ganoderma lucidum extract regulates autophagy induced by MPP⁺ in Neuro-2a cells

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