© 2020 John and Chandrapragasam. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial-
ShareAlike Unported License (http://creativecommons.org/licenses/by-nc-sa/3.0/).
In vitro antioxidant activity of Lactobacillus plantarum against
hydrogen peroxide-induced neuronal damage on PC12 cells
Shani Kunjamma John, Vani Chandrapragasam*
Department of Biotechnology, Karunya Institute of Technology and Sciences, Karunya Nagar, Coimbatore, India.
ABSTRACT
Alzheimer’s disease (AD) is a progressive neurodegenerative disease of the central nervous system. Inflammatory
injury and oxidative stress have been a hallmark for AD due to free radical production. The main objective of
the present study is to identify the compounds having good antioxidant properties from Lactobacillus plantarum
and its effect on neuronal cells. L. plantarum strain was cultured in De Man, Rogosa and Sharpe (MRS) medium
supplemented with cinnamon and pantothenic acid (40 µg/ml and 4 µg/ml). Solvent extraction (ethyl acetate,
chloroform, petroleum ether, and hexane) was done in the supernatant and tested for in vitro 2,2-diphenyl-1-2-
picrylhydrazyl (DPPH), H
2
O
2
scavenging activity, and H
2
O
2
-induced cytotoxicity assessment in PC12 cell lines. The
results obtained from the study showed that all the extracts have good antioxidant activity. It was observed that the
stress-induced PC12 cell lines showed maximum protection of cells in the Media D (chloroform) extract, the IC
50
value was recorded as 23.71 µg/ml. Similarly, all solvent extracts showed significant antioxidant activity in DPPH
assay with IC
50
value ranging from 23.59 to 106.8 µg/ml and IC
50
value in H
2
O
2
scavenging assay ranging from
37.39 to 107.7 µg/ml. Referring to the complex multifactorial etiology of AD, the findings from our work exhibited
remarkable potentials of antioxidants activity.
1. INTRODUCTION
Alzheimer’s disease (AD), a significant health complication in the
civilized world, is a standard form of dotage-related neurodegenerative
dementia. It is a progressive, neurodegenerative disease, the genesis
of neurological decline and stands to be an ultimate and eventually
a life-threatening disease unless the death is interceded by another
cause [1]. AD affects mainly the parts of the brain associated with higher
mental functions, specifically the neocortex and hippocampus [2]. AD is
mainly characterized by abnormal deposition of extracellular amyloid-
beta proteins (Aβ) and intracellular neurofibrillary tangles (tau proteins)
and by comprehensive loss of neurons [3]. Etiology of AD is not fully
understood because of the multifactorial mechanisms underlying the
disease, and various studies suggested that free radicals were involved
in the inflammatory injury and oxidative stress in AD [4,3]. The
neuronal cell death is caused by the elevated production of reactive
oxygen species (ROS) and nitrogen species such as hydrogen peroxide
(H
2
O
2
) and nitric oxide (NO) generated by Aβ in microglial cells [5,6].
Reactive oxygen species (ROS) play a key role in the pathogenesis
of several diseases, including AD [7]. The human brain consumes
*Corresponding Author:
Vani Chandrapragasam,
Department of Biotechnology,
Karunya Institute of Technology and Sciences,
Karunya Nagar, Coimbatore, India.
E-mail: vani@karunya.edu
~20% of oxygen supply and is more susceptible to oxidative stress
due to its high-energy consumption and increased oxidative stress has
been reported in AD patients’ brain [8]. Oxidative stress is a condition
caused by the imbalance between the ROS production and antioxidants
levels causing damage to the cells by excessive production of ROS.
ROS modulate the function of biomolecules and may target the cell
substrates resulting in causing protein, DNA, RNA oxidation, or lipid
peroxidation [9].
Lactobacillus plantarum MTCC 1325 strain was reported to have
the competence of producing acetylcholine (Ach) neurotransmitter
through both external and internal pathways and also holds
significant antioxidant activity [10,11]. The studies on gut–brain axis
communication depict that the bacteria (microbiome) present in the
gastrointestinal tract possibly communicate with the brain and nervous
system in different ways [12].
Pantothenic acid (Vitamin B5) is a water-soluble vitamin, with the
main position for metabolic reactions due to its incorporation into
coenzyme A (CoA) and intake of pantothenic acid is reported for the
reduction of oxidative stress and improved brain damage caused in
gamma-irradiated rats [13,14]. Cinnamon (Cinnamomum zeylanicum
and Cinnamon cassia), known as the eternal tree of tropical medicine,
belongs to the family Lauraceae, besides its antioxidant properties,
they are also recorded to have anti-inflammatory, antimicrobial,
antidiabetic, anticancer, and lipid-lowering and cardiovascular
disease-lowering properties. They are also reported to have activity
Journal of Applied Biology & Biotechnology Vol. 8(05), pp. 84-87, Sep-Oct, 2020
Available online at http://www.jabonline.in
DOI: 10.7324/JABB.2020.80511
ARTICLE INFO
Article history:
Received on: April 23, 2020
Accepted on: August 09, 2020
Available online: September 12, 2020
Key words:
Alzheimer’s disease,
Lactobacillus plantarum,
DPPH scavenging assay,
H
2
O
2
scavenging assay,
PC12 cell line