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Please cite this article in press as Mehri Khatami et al Evaluation of Antioxidant Properties Cherry Fruit
Extract on Prevention the Failure of DNA, Indo Am. J. P. Sci, 2017; 4(07).
INTRODUCTION:
Antioxidants are substances that prevent the synthetic antioxidants have toxic and carcinogenic
formation of free radicals in the cells. Antioxidants effects on human health. Natural antioxidants, such
play an important role in preventing cancer. One of as L-ascorbic acid, are widely used for immunity, but
the most important natural sources of antioxidants is have less antioxidant activity than synthetic
medicinal plants. Medicinal plants are rich in antioxidants. A lot of attention has been drawn in
phenolic compounds. The most important sources of recent decades to finding natural ingredients in foods
natural antioxidants in the human diet are legumes, or herbs to replace synthetic antioxidants. On the
plants and fruits. Natural antioxidants that are present other hand, polyphenols, which are used as natural
in these sources protect the human body from free antioxidants, have become very important because of
radicals and oxidative stress. These antioxidants play their health benefits for humans to reduce the risk of
an important role in human health. Various studies cardiovascular disease and disease-prone illness. [7].
have found that a diet rich in fruits, vegetables, or Antioxidants can purify or inhibit active oxygen
legumes plays an essential role in controlling chronic species by using their alienating properties,
diseases such as heart disease and certain cancers preventing damage by binding to metal ions and
through the freezing of free radicals. Some herbs inhibiting hydroxyl production, as well as lipid
have been reported to contain antioxidant compounds hyperoxidation and DNA damage. There are many
such as bioflavonoids, proanthocyanidins, antioxidants in nature that are different in terms of
phytoestrogens, which have shown protective and physical and chemical structure. They include
anti-cancer properties. It has been shown that these enzymes such as catalase and peroxidases, high
compounds inhibit oxidative stress and molecular weight compounds such as albumin, low
carcinogenesis through several mechanisms. [10]. in molecular weight compounds such as tocopherols,
some plants, it has been shown that phenolic and the latter include plant compounds. [6]
compounds inhibit lipid peroxidation and oxidative
damage to DNA by purifying ROSs such as hydrogen Cherry Extract
peroxide and hydroxyl radicals. There are two main Many herbal compounds and extracts have been used
mechanisms for dealing with free radicals, one of as antioxidants. In addition, there are many edible
which is the non-enzymatic antioxidant system, such fruits that are used as natural and oral antioxidants.
as polyphenols, ascorbic acid and carotenoids.[4] For example, grapevine extract, pomegranate, lemon
Exposure to various chemical and structural and sour cherry are among the most important natural
contaminants such as herbicides, insecticides, toxic antioxidants, with numerous studies on their
chemical waste, cigarette smoke, stress, sunlight and properties. [6] Cherry is one of the fruits that is
diet have similar effects on human health. [14] These predominantly fresh in markets around the world.
contaminations ultimately lead to the oxidation of Cherry is a good source of phenolic compounds that
lipids, proteins, and DNA, which in some cases are considered to be health. Ascorbic acid,
activates carcinogens and leads to cancer, and in flavonoids, and especially anthocyanins are the main
some cases, is associated with aging of the cell and components of the cherry extract, which have a great
other malformations. Oxidative stress plays a role in potential for protecting human health. Anthocyanins
neurodegenerative diseases such as Parkinson, in cherries are found mainly in the skin. Cranberry
Alzheimer and Huntington, as well as multiple products have recently attracted much attention
sclerosis. Examining biomarkers like reactive oxygen because of its beneficial effects on health such as
species and nitrogen or RNS, as well as antioxidant antioxidant activity, nerve cell defense and cancer-
systems, shows that oxidative damage is involved in control properties. These effects are due to its
the pathogenesis of these abnormalities. Oxidative antioxidant compounds, which play an important role
stress is associated with some cardiovascular disease in reducing the oxidative damage and free radicals.
as well as the progression of various cancers. The Several studies have linked cancer inhibition to its
genome of cells is constantly being damaged, which antioxidant effects. [5, 9] Free radicals can lead to
is inevitable due to DNA damage caused by natural many heart diseases such as atherosclerosis, aging
cellular processes. By-products of cell metabolism and cancer. These diseases result from irregular ROS
such as ROS can damage the DNA region and inhibit production and an unbalanced mechanism of
replication. In order to avoid any disruption or antioxidant system protection. To reduce the harmful
damage to the cell, this damage should be prevented effects of ROS, you can use natural antioxidants from
and resolved. Active species produced in oxidative plants and synthetic antioxidants. However, the use
stress cause damage to DNA and are therefore of these compounds has some dangers.
mutagenic, which can cause cell death, proliferation,
invasion and metastasis. [3] Currently, many MATERIALS AND METHODS:
PCR was used in this research. For PCR, it is other uncertain volumes, which facilitates the PCR
necessary to mix DNA patterns, primers, and reaction. The final volume of the PCR mixture should
materials needed to perform a PCR reaction in a be 25 l, so after mixing the material with the
microtiful. The materials used for the PCR reaction appropriate volume, the remaining volume was added
can be used either individually or as a Master Mix to reach 25 l of distilled water. The PCR mixture
kit. In this study, Master Mix [Yekta Takiz Azma] was prepared for each sample according to the
was used. The kit contains materials such as the Taq following table1.
polymerase enzyme, dNTP, PCR buffer, MgCI2 and
EDTA [10mM] -
Phosphate buffer 0.1M and PH=7.4
Cranberry juice
Final volume
Table 4: Final concentration of the required material for Fenton digestive reaction in PCR
Sample number 1 2
3 4 5
Materials required ]Control[
mM] - 5mM 5mM 5mM 5mM
All the steps of mixing the ingredients for PCR PCR mixtures were also closed after tight
should be performed on the ice to prevent the early preparation, and then the specimens were placed in a
activation of the Taq polymerase enzyme and the thermo cycler and, according to the program given to
creation of side products after pouring primers. And the thermo cycler, The PCR cycles were performed
template DNA. Also, all the tools and materials and the DNA pattern was replicated to a large extent
needed to perform PCR should be sterile so that there and used for further experiments.
is no problem in the reaction due to contamination.
The PCR mixtures were also closed after tight Exercise the antioxidant effect of cherry juice on
preparation the samples were then placed in a thermo inhibition of DNA fragmentation by re-PCR
cycler and, according to the program given to the First, the PCR product was diluted 20 times.
thermocycler machine, PCR cycles were performed Cranberry juice was also 5 times diluted and
and the DNA pattern was replicated high and used for according to Table 3, control samples and other
further experiments. samples At 5 micrometers, 1.5 separate, prepared for
Agarose gel electrophoresis incubation at 37 for two hours. After digestion,
Electrophoresis of the agarose gel is required to each sample was diluted 40 times. Then for these
determine the correctness of the PCR and samples, with the same previous PCR program, but
amplification of the desired part. this time With 20 cycles, PCR reaction was
Perform PCR reaction performed again. The agarose gel was then loaded at
All of the PCR reactions were performed in a volume a concentration of 1.5% and electrophoresed at 100
of 25 l with a Peqlab device. volts. The final concentration of substances is given
Perform PCR Reaction in the table 3.
In order to investigate the effect of cherry extract on
preventing DNA fragmentation, digestion product RESULTS:
was used in PCR reaction. To do this, in the 5 new The result of electrophoresis of the PCR of the NOS3
microtitudes, we added the ingredients for the PCR gene
reaction in the table below. We added 3 microlitres of DNA was extracted from four healthy human blood
20-fold diluted digestion to each of the microtutes. samples, NOS3 gene was amplified by PCR and
All steps for mixing the ingredients for PCR should finally electrophoresed The result of electrophoresis
be performed on the ice so that the early activation of is shown in Fig. 1 The DNA Ladder was used to
the Taq polymerase enzyme and the creation of side determine the size of the amplified DNA bands. The
products after pouring out the primers and pattern resulting Sharp bands were found to be free of any
DNA. Also, all the tools and materials needed to excess bandages. The combination of this product
perform PCR should be sterile so that there is no was used for further experiments.
problem in the reaction due to contamination. The
Fig 1- Electrophoresis image of the PCR product of the NOS3 gene. Lines of 1 to 7 blood samples from
healthy individuals.
L line: DNA LADDER [bP 100]
the intensity of the bands on the gel. The results of the 3.Bobe G, Wang B, Seeram NP, Nair MG, Bourquin
third experiment show that up to 4 l of cherry juice is LD. Dietary anthocyanin-rich tart cherry extract inhibits
added; there is a direct relationship between the intestinal tumorigenesis in APCMin mice fed
concentration of cherry juice and the control of the suboptimal levels of sulindac. Journal of agricultural
Fenton reaction. Thus, with increasing cranberry juice and food chemistry. 2006;54[25]:9322-8.
concentrations, inhibition of DNA fracture induced by 4.Das DK. [40] Naturally occurring flavonoids:
Fenton reaction was further increased, but an inverse Structure, chemistry, and high-performance liquid
relationship was observed at higher concentrations. We chromatography methods for separation and
conclude that according to recent studies, the higher the characterization. Methods in enzymology.
antioxidant concentration used in the reaction, the 1994;234:410-20.
greater the inhibition of DNA, and thus the direct 5.Faria-A, Monteiro R, Mateus M, Izevedo I, CAlhau C.
relationship between the concentration of cherry juice Effect of pomegranate[Punica granatum] juice intake on
and the control of the Fenton reaction is justified. hepatic oxidativ stress.Eurpuropeam Jour nal of
Nutritiom.2007:46[5]:271 8.
CONCLUSION: 6.Jovanovic SV, Steenken S, Tosic M, Marjanovic B,
Extract of antioxidant-rich plants, if used as a brain Simic MG. Flavonoids as antioxidants. Journal of the
substance, reduces oxidative stress and, consequently, American Chemical Society. 1994;116[11]:4846-51.
the risk of neurodegenerative diseases. Polyphenols are 7.Kurek-Grecka A, Rzepecka-Stojko A, Grecki M,
among the compounds that have antioxidant properties. Stojko J, Sosada M, wierczek-Ziba G. Structure and
Feeding polyphenols can protect the cell against antioxidant activity of polyphenols derived from
oxidative damage. Cherry is rich in phenolic propolis. Molecules. 2013;19[1]:78-101.
compounds. This study expresses the antioxidant and 8.Mertens Talcott SU, Jilma Stohlawetz P, Rios J,
cherry juice protection of genomic DNA and the Hingorani L, Derendorf H. Absorptiom, metabolism,
pBR322 plasmid against damage. . These properties of and antioxidant effects of pomegranate[Punica granatum
cherry juice, it introduces it as a source of natural L.] polyphenols after ing estion of a standardrzde extract
antioxidants with the potential for reducing oxidative in healthy human volunteers. Journal of Agricuitural and
stress and ultimately maintaining health. According to Food chemistry. 2006: 54[23]: 8956- 61.
the results of this study, the best concentrations of 9.Lopes GK, Schulman HM, Hermes Lima M.
cherry juice were obtained in order to prevent genomic Polyphenol tannic acid inhibits hydroxyl radical
formation from Fenton reaction by complexing ferrous
DNA fragmentation at and hours, and at 15 to 15 ions. Biochimica et Biophysica Acta[BBA] General
hours, the best concentrations of cherry juice were Subjects.1992:1472[1]:142-52.
obtained to prevent plasmid defeat the pBR 322 was 10.Podmore ID, Griffiths HR, Herbert KE, Mistry N,
10% vol / vol. Mistry P, Lunec J. Vitamin C exhibits pro-oxidant
properties. Nature. 1998;392[6676]:559-.
ACKNOWLEDGMENT 11.Pandey KB, Rizvi SI. Plant polyphenols as dietary
This article is extracted from my thesis under the title of antioxidants in human health and disease. Oxidative
Evaluation of antioxidant properties Cherry fruit medicine and cellular longevity. 2009;2[5]:270-8.
extract on prevention the failure of DNA. Hereby, I 12.Richardson DR, Lok HC. the nitric oxide iron
extend my sincere appreciation to Islamic Azad interplay in mammalian cells:Transport and storage of
university of, Ashkezar for the efforts and supports they dinitrosyl iron complexes. Biochimica et Biophysica
provided to me. Icta [BBA] General Subjects. 2008:1780[4]: 638-51.
13.Winterbourn CC. the ability of scavengers to
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