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MEDICINAL ANALYSIS OF THE PLANT GLYCYRRHIZA GLABRA
Authors:
Safokulov Bobur
3rd year student of Clinical Pharmacy,
Tashkent Pharmaceutical Institute
Khusankhonov Azizkhon
1st year student of the Industrial
Pharmacy department of the Tashkent
Pharmaceutical Institute
Ayozov Shohabbos
3rd year student of the Pharmaceutical
Analysis Department of the Tashkent
Pharmaceutical Institute
Keywords:
Antimicrobial, antioxidant, antiproliferative, Glycyrrhiza glabra,
medicinal plants
Abstract
Objective: Alternative medicine has an important place in the fight against many
diseases in human history. The biological activity of Glycyrrhiza glabra L. was
investigated in this study. Material and Method: The root parts of the plant were
extracted with ethanol. Antioxidant and oxidant potentials were determined using Rel
Assay kits. Antimicrobial activity was tested against standard bacteria and fungus
strains using the agar dilution method. Antiproliferative activity was determined by
MTT test against Lung Carcinoma Cell Line (A549). Result and Discussion: As a result
of the studies, the TAS value of the plant was measured as 8.770±0.171, TOS value as
14.590±0.191 and OSI value as 0.167±0.005. Inhibition of the plant extract was
observed against standard bacteria and fungus strains at ranging from 50-200 µg/mL
concentrations. In addition, it was determined that the plant extract displayed strong
antiproliferative activity due to the increase in concentration. As a result of these
studies, it has been determined that G. glabra can be used as an important natural
antioxidant, antimicrobial and anticancer agent.
INTRODUCTION
Many natural materials such as mushrooms, plants and animals are used in
alternative medicine [1]. It occupies a very important place in the treatment of diseases
due to the active substances produced by the plants. Many studies have shown that
plants have anticancer, antioxidant, antimicrobial, antiproliferative, anti-inflammatory,
DNA protective, antiallergic and hepatoprotective activities [2-4]. In our study,
Glycyrrhiza glabra L. (Licorice) was used as a material. Licorice; It is a plant belonging
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to the genus Glycyrrhiza glabra, of the genus Glycyrrhiza of the Papillionacease family.
It is used for therapeutic purposes known in the history of ancient medicine in many
civilizations such as Sumer, Mesopotamia, China, Greek and Egypt in the world.
Among the aromatic and medicinal plants, the licorice plant is popularly known by 14
different names such as "biyam, dye, piyam, sweet root, etc." [5]. Licorice plant in the
world at 12, while in Turkey in 6 species of yellow-blue or brown color, is a perennial
shrub plant ranging between 30-160 cm in length [6]. The roots of the licorice plant are
biologically known as a source of magnesium and silicon. In addition, the active
ingredient in the composition of the licorice plant, glycyrrhizin is 50 times sweeter than
tea sugar and 150 times sweeter than sucrose. Glycyrrhiza acid, which is found in the
sweetness of the roots, and its calcium and potassium salts, two of the substances such
as sucrose and mannite come from [7, 8]. Therefore, as much more intense taste is
obtained with less amount, it has been involved in the cuisine and food industry of
many countries for centuries [9]. In addition, in the production of licorice honey,
licorice sherbet, in the manufacture of tobacco, snuff and filter cigarettes, in the
confectionery and beverage industry as a fragrance and flavoring, in cosmetics, velvet
dyeing and shoe dyeing in the textile industry, making foam in fire fighting, in
preparations prepared to kill insults, and in the food industry, there are areas of use
such as adding fragrance to foods [10-12]. Licorice plant is widely used in food,
confectionery, medicine and tobacco products as a flavoring agent known worldwide
as "generally safe" (GRAS) [13]. In this study, antioxidant, oxidant, antimicrobial and
antiproliferative activity of ethanol extract of root parts of G. glabra was determined.
MATERIAL AND METHOD
Laboratory Studies
Plant samples were collected from Duhok (Iraq). Soil and dust particles were
removed from the root parts of the plant. It was then dried under suitable conditions.
After drying the plant parts were pulverized and weighed 30 g. It was then extracted
with ethanol for about 6 hours, for example at 50 °C. The solvents of the extracts were
removed in a rotary evaporator and crude extracts were obtained.
Antioxidant Parameters
The antioxidant and oxidant status of the plant extract was determined using Rel
Assay TAS and TOS kits. TAS tests were performed according to the protocol
specified in Erel [14] and Trolox was used as a calibrator. Results are shown in mmol
Trolox equiv./L. TOS tests were performed according to the protocol specified in Erel
[15] and hydrogen peroxide was used as a calibrator. Results are shown as μmol H2O2
equiv./L. The oxidative stress index (OSI) was determined by proportioning the TOS
value to the TAS value [16].
Antimicrobial Activity Tests
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The antimicrobial activity of the root parts of the plant against EtOH extract
bacteria and fungus strains was determined using the agar dilution method [17-19]. The
plant extract was adjusted with distilled water at 800-12.5 µg/mL concentrations.
Bacterial strains were set in Muller Hinton Broth medium. Staphylococcus aureus
ATCC 29213, S. aureus MRSA ATCC 43300, Enterococcus faecalis ATCC 29212,
Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and
Acinetobacter baumannii ATCC 19606 were used as bacterial strains. Fungus strains
were pre-cultured in RPMI 1640 Broth medium. Candida albicans ATCC 10231, C.
krusei ATCC 34135 and C. glabrata ATCC 90030 were used as fungus strains. The
extract concentration that inhibits the growth of bacteria and fungus strains was
determined as the MIC value. Results were expressed in µg/mL [20-22].
Antiproliferative Activity Tests
The antiproliferative activity of the EtOH extract of the plant was determined by
MTT test on A549 lung cancer cells. Cells were separated after 70-80% confluence
using 3.0 mL of Trypsin-EDTA solution (Sigma-Aldrich, MO, USA). It was then
planted on plates and incubated for 24 hours. The plant extract (25, 50, 100, 200
µg/mL) was then adjusted at different concentrations. After the incubation period, the
supernatants were dissolved in growth medium and replaced with 1 mg/mL MTT
(Sigma). It was then incubated at 37 °C until a purple precipitate formed. The
supernatants were then removed and dissolved by adding dimethyl sulfoxide (DMSO)
(Sigma-Aldrich, MO, USA) to MTT absorbed by cells. Subsequently, plates were read
at 570 nm using an Epoch spectrophotometer (BioTek Instruments, Winooska, VT)
[23].
RESULT AND DISCUSSION
Antioxidant Activity In recent years, it is known that the basis of many diseases
are due to oxidative stress caused by reactive oxygen species. Oxidative stress is due
to the disproportion between the formation and neutralization of prooxidants [24]. The
level of endogenous oxidant compounds resulting from environmental factors as a
result of metabolic activities is highly toxic when they accumulate in organisms. The
antioxidant defense system plays a role in reducing the effects of oxidant compounds.
If the antioxidant defense system is insufficient, oxidative stress occurs. In such cases,
supplemental antioxidants are important in reducing the effect of oxidative stress. In
this context, it is very important to identify new natural antioxidant agents [25]
In previous studies of G. glabra, it was reported that aqueous, methanol, ethanol
extracts have antioxidant potential using different methods (Inhibition of b-carotene–
linoleate bleaching, Hypochlorous acid-scavenging activity, Inhibition of
myeloperoxidase-chlorinating system, Nitric oxide radical scavenging activity,
Superoxide anion scavenging activity, Hydroxyl radical scavenging activity, DPPH
radical scavenging activity, ABTS•+ cation Radical Scavenging, Fe+2/ascorbate
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induced lipid peroxidation assay, Reducing power) [26-29]. In our study, TAS, TOS
and OSI values were determined for the first time by using Rel Assay kits of G. glabra.
As a result of the studies, it has been determined that G. glabra has an important
antioxidant activity. In addition, TAS values of R. coriaria var. zebaria, M. longifolia
subsp. longifolia, A. calocephalum, S. papposa, F. platycarpa, T. spicata, G.
tournefortii, R. crispus and A. millefolium reported in the literature were reported as
7.342, 3.628, 5.853, 5.314, 5.688, 8.399, 6.831, 6.758 and 2.436 mmol/L, respectively.
TOS values were reported as 5.170, 4.046, 16.288, 24.199, 15.552, 6.530, 3.712, 5.802
and 2.839 µmol/L, respectively. OSI values were reported as 0.071, 0.112, 0.278,
0.456, 0.273, 0.078, 0.054, 0.086 and 0.083, respectively [30-38]. Compared to these
studies, the TAS value of G. glabra was determined to be higher than R. coriaria var.
zebaria, M. longifolia subsp. longifolia, A. calocephalum, S. papposa, F. platycarpa, T.
spicata, G. tournefortii, R. crispus and A. millefolium. TAS value shows all of the
antioxidant compounds produced in the plant [30]. As seen in our study, it has been
determined that G. glabra has a very important antioxidant potential.
When TOS values were examined, it was determined that G. glabra was lower
than S. papposa and F. platycarpa, and higher than Rhus coriaria var. zebaria, Mentha
longifolia subsp. longifolia, A. calocephalum, T. spicata, G. tournefortii, R. crispus and
A. millefolium. The TOS value indicates all of the oxidant compounds produced by
the environmental effects in the plant [30]. It is seen that the oxidant levels of the plant
used in our study are at normal levels. When OSI values were examined, it was
determined that G. glabra was lower than A. calocephalum, S. papposa and F.
platycarpa, and higher than R. coriaria var. zebaria, M. longifolia subsp. longifolia, T.
spicata, G. tournefortii, R. crispus and A. millefolium. The OSI value shows how much
oxidant compounds produced in the plants are suppressed by the antioxidant defense
system. A low OSI value indicates that the antioxidant defense system of the plant
works well [30]. In our study, it was determined that the antioxidant defense system of
G. glabra was sufficient in suppressing oxidant compounds. As a result, it was
determined that G. glabra has significant antioxidant activity.
Antimicrobial Activity
Today, many diseases occur due to microorganisms. Antibiotics are used
extensively in the treatment of microorganism-based diseases. Today, resistant strains
are emerging due to the unconscious use of antibiotics [39]. Antibiotics used against
resistant microorganisms are insufficient. In addition, due to the possible side effects
of chemical antibiotics, the tendency towards natural products is increasing. In this
context, the discovery of new antimicrobial drugs is inevitable [40]. In this study, the
activities of G. glabra against bacteria and fungi were investigated.
In previous studies, it has been reported that methanol extracts of G. glabra are
effective against Staphylococcus aureus, Bacillus megaterium, B. subtilis, Sarcina
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lutea, Salmonella paratyphi, S. typhi, Escherichia coli, Shigella dysenteriae, Vibrio
minicus, V. parahemolyticus and Psedomonas aeruginosa at different concentrations
[41]. In a different study, ethanol extracts of G. glabra were reported to be effective
against Candida albicans, Staphylococcus aureus, Bacillus subtilis, Enterococcus
faecalis, Escherichia coli, Pseudomonas aerouginosa and Klebsiella pneumoniae [42].
Ethanolic, hexane fraction, ethyl acetate fraction and methanol fraction of G. glabra
have been reported to be effective against Staphylococcus aureus, Staphylococcus
epidermidis, Streptococcus mutans, Bacillus subtilis, Enterococcus faecalis, Klebsiella
pneumoniae, Salmonella typhi, Yersinia enterocolitica, Enterobacter aerogens and
Escherichia coli [43]. In our study, G. glabra was determined to be effective against A.
baumannii, C. glabrata and C. albicans at 25 µg/mL, S. aureus, S. aureus MRSA and
C. krusei at 50 µg/mL, E. faecalis and P. aeruginosa at 100 µg/mL, E. coli at 200 µg/mL
extract concentrations. As a result, it was determined that G. glabra has antibacterial
and antifungal activities.
Conclusion
In this study, some medicinal properties of the root parts of G. glabra were
determined. As a result of the studies, it was determined that the root extracts of the
plant exhibit significant antioxidant, antimicrobial and antiproliferative activity. As a
result, it is thought that G. glabra can be used as a natural material in pharmacological
designs.
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