Polydatin protects SH-SY5Y in models of Parkinson's disease by promoting Atg5-mediated but parkin-independent autophagy
Graphical abstract
Schematic diagram underlying the protective effects of polydatin against Parkinson's disease models induced by Rot or Parkin knockdown through recovering redox balance, rescuing autophagy and inhibiting mitochondrial fusion.
Introduction
Parkinson's disease (PD), the second most common neurodegenerative disease, is characterized by a progressive loss of dopaminergic neurons in the substantia nigra, affecting approximately 7–10 million patients worldwide (de Lau and Breteler, 2006). Currently, the pathogenesis of PD is not completely understood. However, recent studies have uncovered some pathogenic mechanisms underlying PD, including oxidative stress, mitochondrial dysfunction, and impaired autophagy (Charvin et al., 2018). Oxidative stress, which is intimately linked to mitochondrial dysfunction and α-synuclein aggregation, is considered to be a potential driver of PD progression (Jenner and Olanow, 1996; Surmeier et al., 2011). Mitochondrial dysfunction via the generation of excessive reactive oxygen species (ROS) has also been implicated in PD pathogenesis (Park et al., 2018). In addition, impairment of autophagy can lead to α-synuclein release and dysfunctional mitochondrial accumulation (Gao et al., 2017). Obviously, the etiology of PD is multifactorial and several putative pathogenetic pathways are intimately linked. New drugs combining the activities of antioxidants, mitochondrial protectors, and autophagy enhancers appear to have great potential in PD treatment (Arduino et al., 2010; Giordano et al., 2014; Filograna et al., 2016).
Recent studies have shown that polyphenols have a great capacity for neuroprotection through diverse potential mechanisms of action—scavenging ROS, affecting mitochondrial functions, and protecting against protein aggregation (Nabavi et al., 2018). Several clinical studies have reported the effects of polyphenols in neurodegenerative pathologies, indicating that these natural molecules can potentially increase memory and cognitive functions (Libro et al., 2016; Vacca et al., 2016; Ajami et al., 2017). The promising pharmacological role of natural polyphenols mainly depends on their bioavailability (Manach et al., 2004). For example, resveratrol, a highly active bioactive compound with multiple beneficial properties, has shown very low bioavailability and a rapid clearance rate (Goldberg et al., 2003; Smoliga and Blanchard, 2014). However, some of its derived metabolites are considered to exert better biological activities. These include polydatin (Pol), a non-glycosylated derivative of resveratrol (Wu et al., 2015). To date, the biological and pharmacological properties of Pol have not been explored in-depth. A study reported that it exhibits potent antioxidant activity and can attenuate dopaminergic neurodegeneration in three commonly used rodent PD models (Chen et al., 2015). Moreover, Pol has been demonstrated to improve learning and memory in rats treated with chronic ethanol exposure (Zhang et al., 2015). This sparked our interest in studying the therapeutic potential of Pol in PD treatment.
In this study, we aimed to evaluate the neuroprotective effect of Pol in human neuroblastoma cells (SH-SY5Y) using both environmental and genetic PD models. Rotenone (Rot), an inhibitor of mitochondrial complex I, has been used to induce PD models by elevating ROS and depleting ATP. In addition, mutations leading to the loss of Parkin protein function account for the most common familial forms of PD with autosomal recessive inheritance (Periquet et al., 2003; Jin and Youle, 2012). Thus far, there have been few reports on the effectiveness of Pol against a Parkin-mediated PD model. We, therefore, focused on investigating the effects of Pol on cell injury and mitochondrial function in both Rot-treated and Parkin knockdown SH-SY5Y cells and elucidating the underlying mechanisms by investigating the involvement of autophagy (mitophagy) signaling pathways. After observing the significant protective effects of Pol in vitro, we used genetic mutant Drosophila to model PD and investigated whether Pol could rescue the phenotypes of parkin-null flies. Collectively, our results suggest that Pol could be developed as a potential candidate for PD treatment.
Section snippets
Chemicals and reagents
Pol (dissolved in distilled water, purity > 98%) was purchased from the Shanghai Yuanye Biotechnology Company (Shanghai, China). Rot and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The dopamine ELISA kit was obtained from the Cloud-Clone Corporation (Wuhan, China). The primary antibodies against SIRT1, DJ1, mTOR, p-mTOR (Ser2488), Ulk, p-Ulk(Ser757), Parkin, PGC1α, and Drp1 were all obtained from Cell Signaling Technology (Beverly, MA, USA). Anti-LC3
Rot-induced injury, oxidative stress, and mitochondrial dysfunction of SH-SY5Y cells
We first evaluated the cytotoxic response of SH-SY5Y cells to Rot. Cells exposed to increasing concentrations of 0.125–0.5 μM Rot for 24 h exhibited progressive cytotoxicity (Fig. 1a and b) without inducing apoptosis (Fig. S1). In the presence of 0.5 μM Rot, only 65.9% viable cells were observed while 195.8% LDH release were detected, compared to the control group. The mitochondria are the target organelles for Rot toxicity. Our data showed that Rot significantly increased ROS levels (Fig. 1c)
Discussion
The evidence for the occurrence of oxidative stress in PD is overwhelming. The central nervous system is very susceptible to ROS because it consumes large amounts of oxygen and is not particularly enriched in antioxidant defenses compared to other tissues (Bhat et al., 2015). . Pol, a natural precursor of resveratrol, has a stronger antioxidative effect compared to resveratrol (Ravagnan et al., 2013). In our study, ROS was increased in Rot-exposed, Parkin knockdown, and Atg5 knockdown SH-SY5Y
Funding
This study was supported by the National Science Foundation of China (81672508), Jiangsu Provincial Foundation for Distinguished Young Scholars (BK20170041), Natural Science Foundation of Shaanxi Province (2019JM-016, 2019JM-130), China-Sweden Joint Mobility Project (51811530018), and Fundamental Research Funds for the Central Universities.
CRediT authorship contribution statement
Hua Bai: Conceptualization, Methodology, Investigation, Writing - original draft. Yaqi Ding: Investigation, Methodology, Writing - original draft. Xin Li: Formal analysis, Visualization, Data curation. Deqin Kong: Investigation, Visualization, Data curation. Chenqi Xin: Software, Validation. Xuekang Yang: Resources. Chengwu Zhang: Project administration. Ziqiang Rong: Resources. Chuanhao Yao: Writing - review & editing. Shenci Lu: Writing - review & editing. Lei Ji: Resources. Lin Li:
Declaration of competing interest
The authors do not have any conflicts to disclose.
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These authors contributed equally.