Bindhu, J (2019) Anthraquinone from Edible Fungi Pleurotus ostreatus Protects Human SH-SY5Y Neuroblastoma Cells Against 6-Hydroxydopamine-Induced Cell Death—Preclinical Validation of Gene Knockout Possibilities of PARK7, PINK1, and SNCA1 Using CRISPR SpCas9. Applied Biochemistry and Biotechnology.

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Abstract

Parkinson’s disease (PD) results from the degeneration of the nervous tissue brought
about by ecological and hereditary components which affects nerve cells in the brain. It is
the world’s second most normal neurodegenerative issue, which can essentially weaken
the personal satisfaction, make reliance, and trigger untimely mortality of affected people.
The commonness pace of PD is 0.5–1% among individuals in the age group of 65–69
years and 1–3% among those 80 or more. Clinical appearances incorporate bradykinesia,
tremors, unbending nature, and postural unsteadiness; spectrums of non-motor symptoms
include psychological hindrance and passionate and behavioral brokenness. In this study,
6-OHDA-induced neurotoxicity was analyzed for various cytotoxicity analyses. The
genes identified were PINK1 (PTEN-induced kinase 1), PARK7 (Parkinsonismassociated deglycase) and SNCA 1 (alpha synuclein1) validated using CRISPR spcas9
genome editing tool. In this study, Anthraquinone isolated from Pleurotus ostreatus was
treated against a dopaminergic neurotoxin, 6-hydroxydopamine (6-OHDA), which induced neurotoxicity in SH-SY5Y cells. Experimental groups in SH-SY5Y neuroblastoma
cells were treated with anthraquinone (50 nM) and 6-OHDA (100 nM). MTT and ROS
assays were performed to assess the cell viability and oxidative stress within the cells,
followed by mixed-member proportional (Mitochondrial membrane potential), dual
staining, and immunoblotting. 6-OHDA-induced cell death in SH-SY5Y cells was
dose-dependently attenuated by treatment with anthraquinone. The genes responsible
for mutation were studied and the mutated RNAs knockout possibilities was studied
using CRISPR spcas9 genome editing tool. Treatment with anthraquinone attenuated the
level of oxidative stress and reduced the mitochondrial dysfunction associated with 6-
OHDA treatment. Immunoblot analysis carried out with apoptotic markers showed that
cytochrome C and caspase-3 expression increased significantly in anthraquinone-treated
cells, whereas 6-OHDA-treated group showed a significant decrease when compared with
an experimental control group. The mutated genes PARK7, PINK1, and SNCA1 were
analyzed and found to exhibit four gene knock possibilities to treat PD. Reports demonstrate that other than following up on the biosynthesis of dopamine and its metabolites,these mixes counteract D2 receptors’ extreme touchiness. It is proposed that further
examinations need be directed to better understand the activity of the bioactive mixes
circulated in these edible fungi Pleurotus ostreatus. The gene knockout possibilities
identified by CRISPR SpCas9 will pave a way for better research for PD treatment.

Item Type: Article
Uncontrolled Keywords: Anthraquinone . SH-SY5Y. neuroblastoma . Dopamine . 6-OHDA . CRISPR
Divisions: PSG College of Arts and Science > Department of Biotechnology
Depositing User: Mr Team Mosys
Date Deposited: 03 Aug 2022 08:30
Last Modified: 03 Aug 2022 08:30
URI: http://ir.psgcas.ac.in/id/eprint/1403

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