Abstract
Neurodegenerative diseases (NDDs) are chronic conditions that have drawn robust interest from the scientific community. Phytotherapeutic agents are becoming an important source of chemicals for the treatment and management of NDDs. Various secondary metabolites have been isolated from Melicope lunu-ankenda plant leaves, including phenolic acid derivatives. However, their neuroprotective activity remains unclear. Thus, the aim of this study is to elucidate the in vitro neuroprotective activity of 7-geranyloxycinnamic acid isolated from Melicope lunu-ankenda leaves. The neuroprotective activity was evaluated in differentiated human neuroblastoma (SH-SY5Y) cells by monitoring cell viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Moreover, the potential to impair apoptosis in differentiated cells was investigated employing the Annexin V-FITC assay, acridine orange and propidium iodide (AO/PI) staining, and fluorescence microscopy. Morphological assessment and ultrastructural analysis were performed using scanning and transmission electron microscopy to evaluate the effect of 7-geranyloxycinnamic acid on surface morphology and internal features of the differentiated cells. Pre-treatment of neuronal cells with 7-geranyloxycinnamic acid significantly protected the differentiated SH-SY5Y cells against H2O2-induced apoptosis. Cytoskeleton and cytoplasmic inclusion were similarly protected by the 7-geranyloxycinnamic acid treatment. The present findings demonstrate the neuroprotective potential of 7-geranyloxycinnamic acid against H2O2-induced neurotoxicity in neuronal cells, which is an established hallmark of neuronal disorders.
Highlights
Neurodegenerative diseases including Alzheimer’s disease (AD), Huntington’s disease (HD), Parkinson’s disease (PD) and amyotrophic lateral sclerosis (ALS) are distinguished by the slow loss of Molecules 2020, 25, 3724; doi:10.3390/molecules25163724 www.mdpi.com/journal/molecules Molecules25, x FOR PEER REVIEW 2 of 172 of 17
The highest viability was noted when the cells were treated with 2.08 μM 7-geranyloxycinnamic acid, after 72 h of treatment (Figure 5A), cells and its ability to inhibit the cytotoxicity induced by H2O2 were investigated
The cells were incubated for 24 h at 37 ◦ C in a 5% CO2 incubator, and differentiated by addition of 10 μM all trans retinoic acid (RA), dissolved in DMEM/F12 media supplemented with 3% heat-inactivated fetal bovine serum (FBS), and further incubated in the dark at 37 ◦ C in a 5% CO2 incubator for 7 days
Summary
Neurodegenerative diseases including Alzheimer’s disease (AD), Huntington’s disease (HD), Parkinson’s disease (PD) and amyotrophic lateral sclerosis (ALS) are distinguished by the slow loss of Molecules 2020, 25, 3724; doi:10.3390/molecules25163724 www.mdpi.com/journal/molecules. 1. Introduction neuronal function [1,2], and their pathogenesis has been linked to oxidative stress, a condition induced by excessive production of reactive oxygen species (ROS) including hydrogen peroxide (H2 O2 ) [3,4]. Increased oxidative stress elevation is seen in the brain of patients system gives rise to oxidative injury of biological molecules, resulting in the initiation of the processes with neuronal disorders [9,10]. Melicope including lunu-ankenda (Rutaceae), known locally in Malaysia as “tenggek burung” Such asfrom polyphenols hydroxycinnamic acid derivatives and hydroxybenzoic burung”. Polyphenols, including phenolic acids, have been neuroprotective activity through the prevention of protein oxidation, ROS generation and lipid demonstrated to exert neuroprotective activity through the prevention of protein oxidation, ROS peroxidation as well as anti-apoptotic and anti-inflammatory activities [18,19,20].
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