Larrea tridentata Extract Mitigates Oxidative Stress-Induced Cytotoxicity in Human Neuroblastoma SH-SY5Y Cells.

Karla Morán-Santibañez,Rachid Skouta,Karine Fenelon,Abimael H Vasquez,Veronica Henderson,Janae Sweeney,Valerie Odero-Marah,Armando Varela-Ramirez

doi:10.3390/antiox8100427

Abstract

Creosote bush (Larrea tridentata; LT) leaves extracts were tested for their potential efficacy to mitigate cellular oxidative stress on human SH-SY5Y cells. Here, the differential nuclear staining assay, a bioimager system, and flow cytometric protocols, concurrently with several specific chemicals, were used to measure the percentage of cell viability and several facets implicated in the cytoprotective mechanism of LT extracts. Initially, three LT extracts, prepared with different solvents, ethanol, ethanol:water (e/w), and water, were tested for their capacity to rescue the viability of cells undergoing aggressive H2O2-induced oxidative stress. Results indicate that the LT extract prepared with a mixture of ethanol:water (LT-e/w; 60:40% v/v) displayed the most effective cytoprotection rescue activity. Interestingly, by investigating the LT-e/w mechanism of action, it was found that LT-e/w extract decreases the levels of H2O2-provoked reactive oxidative species (ROS) accumulation, mitochondrial depolarization, phosphatidylserine externalization, caspase-3/7 activation, and poly (ADP-ribose) polymerase (PARP) cleavage significantly, which are hallmarks of apoptosis. Thus, out of the three LT extracts tested, our findings highlight that the LT-e/w extract was the most effective protective reagent on SH-SY5Y cells undergoing oxidative stress in vitro, functioning as a natural anti-apoptotic extract. These findings warrant further LT-e/w extract examination in a holistic context.

Highlights

In a previously published study, we evaluated the antioxidant activity of extracts of Larrea tridentata, a plant commonly used in traditional medicine for its antiviral, antiparasitic, anti-inflammatory, and antioxidant properties [18,19,20]
To optimize the incubation time and concentrations utilized in the cytotoxicity rescue experiments, either Larrea tridentata (LT) extracts or H2 O2 several were tested independently on SH-SY5Y cells, measuring their viability using differential nuclear staining (DNS) assay and a bioimager system
From these series of experiments, cells treated for 12 h with 600 μM of H2 O2 were selected for subsequent rescue experiments, as the percentage of its cytotoxicity observed was around 50%, as compared with solvent-treated cells (Figure 1B)

Summary

Introduction

Neurodegenerative diseases (NDs) are characterized by progressive deterioration of the structure and function of the central nervous system that leads to compromise motor or cognitive function [1,2].Oxidative stress due to the excessive production of reactive species, especially reactive oxygen (ROS)and nitrogen (RNS) species besides the failure of balance by antioxidant enzyme systems, have been involved in the development of Alzheimer’s disease (AD), Parkinson’s disease (PD), and other NDs [3,4].Antioxidants 2019, 8, 427; doi:10.3390/antiox8100427 www.mdpi.com/journal/antioxidantsOxidative damage in the cell manages the destruction of biomolecules such as lipids, proteins, and DNA, so counteracting the effects produced by this oxidative stress is one of the approaches to neuroprotection [5]. Neurodegenerative diseases (NDs) are characterized by progressive deterioration of the structure and function of the central nervous system that leads to compromise motor or cognitive function [1,2]. Oxidative stress due to the excessive production of reactive species, especially reactive oxygen (ROS). Neuroprotection can be defined as the relative strategies and mechanisms that defend the central nervous system for neuronal injuries due to both acute (e.g., stroke or trauma) and chronic neurodegenerative diseases [6]. The lack of successful therapies to overcome oxidative stress without side effects has prompted the research of new treatments. The main approaches of new therapies have focused on three main sources: Synthesis of new drugs, drug repurposing, and natural products research [7,8]. Plant extracts have been specially studied and proven to be effective for memory enhancement, anti-aging, and neuroprotective purposes [9,10,11]

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