Received: 19/08/2024 Accepted: 04/11/2024 Published: 20/01/2025 1 of 7
https://doi.org/10.52973/rcfcv-e35502 Revista Cientíca, FCV-LUZ / Vol. XXXV
ABSTRACT
This study aimed to view the antioxidant and anti–inflammatory
properties of Helichrysum plicatum DC. subsp. plicatum (HP)
methanol extract on the urinary tract using an experimentally
induced urolithiasis (U) model. The study included four groups:
Group 1 was given a standard diet, Group 2 was given a diet
added with HP, Group 3 was given a standard diet with induced
urolithiasis, and Group 4 received an HP–supplemented diet with
induced urolithiasis. Rats in Groups 2 and 4 were administered
500 mg·kg
-1
·day
-1
of HP via gavage feeding for 21 days. Urolithiasis
was induced in Groups 3 and 4 by administering 1% ethylene
glycol and 1% ammonium chloride in their swig water for 21 days
to create a calcium oxalate (CaOx) urolithiasis model. The study
analyzed plasma concentrations of thiobarbituric acid reactive
substances (TBARS), an indicator of serum oxidative stress (OS),
HP, and HP levels. Additionally, oxalate (Ox), urea, calcium, and
creatinine clearance levels were measured in both blood and urine,
and routine histological evaluations were conducted. The results
indicated important higher concentrations of HP in the groups given
HP (P<0.001), while plasma TBARS concentrations were lower
in Group 4 compared to Group 3 (P=0.001). The ndings suggest
that HP reduces OS by lowering plasma TBARS levels induced by
CaOx, due to its antioxidant and anti–inflammatory properties.
Furthermore, the measured biochemical measurements supported
the anti–urolithiasis effects of HP. In results, this study supports
the hypothesis that HP’s antioxidative and anti–inflammatory
properties help prevent OS, which is a factor in stone formation,
thereby preventing acute renal damage and stone formation.
Key words: Calcium oxalate crystals; urolithiasis; rat; thiobarbituric
acid reactive substance (TBARS); Helichrysum
plicatum DC. subsp. plicatum
RESUMEN
Este estudio tuvo como objetivo evaluar las propiedades
antioxidantes y antiinflamatorias del extracto de metanol de
Helichrysum plicatum DC. subsp. plicatum (EP) en el tracto urinario,
utilizando un modelo de urolitiasis (U) inducido experimentalmente.
El estudio incluyó cuatro grupos: al Grupo 1 se le administró una
dieta estándar convencional, al Grupo 2 se le administró una dieta
complementada con EP, al Grupo 3 se le administró una dieta
estándar con urolitiasis inducida y el Grupo 4 recibió una dieta
complementada con EP y con urolitiasis inducida. A las ratas de los
Grupos 2 y 4 se les administró 500 mg·kg
-1
·dia
-1
de EP mediante
alimentación por sonda durante 21 dias. La urolitiasis se indujo en
los Grupos 3 y 4 mediante la administración de 1 % de etilenglicol
y 1 % de cloruro de amonio en su agua potable durante 21 dias
para crear un modelo de urolitiasis por oxalato de calcio (CaOx).
El estudio analizó las concentraciones plasmáticas de sustancias
reactivas al ácido tiobarbitúrico (TBARS), un indicador de estrés
oxidativo (EO), así como los niveles de EP en el suero. Además, se
midieron los niveles de oxalato (Ox), urea, calcio y la depuración
de creatinina en sangre y orina, y se realizaron evaluaciones
histológicas de rutina. Los resultados indicaron concentraciones
signicativamente más altas de EP en los grupos que recibieron
EP (P<0,001), mientras que las concentraciones plasmáticas
de TBARS fueron más bajas en el Grupo 4 en comparación con
el Grupo 3 (P=0,001). Los hallazgos sugieren que EP reduce el
EO al disminuir los niveles plasmáticos de TBARS inducidos por
CaOx, debido a sus propiedades antioxidantes y antiinflamatorias.
Además, las mediciones bioquímicas realizadas respaldaron los
efectos antiurolíticos de EP. En resumen, este estudio respalda la
hipótesis de que las propiedades antioxidantes y antiinflamatorias
de EP ayudan a prevenir el EO, que es un factor en la formación
de cálculos, previniendo así el daño renal agudo y la formación
de cálculos.
Palabras clave: Cristales de oxalato de calcio; urolitiasis; rata;
sustancia reactiva al ácido tiobarbitúrico (TBARS);
Helichrysum plicatum DC. subsp. plicatum
Antioxidant and anti–inflammatory effects of Helichrysum plicatum DC.
subsp. plicatum extract in an experimental model of acute urolithiasis
Efectos antioxidantes y antiinflamatorios del extracto de Helichrysum plicatum
DC. subsp. plicatum, en un modelo experimental de urolitiasis aguda
Selvinaz Yakan
1
* , Kıvılcım Eren Erdoğan
2
, Yusuf Kenan Dağlıoğlu
3
, Tuba Aydın
4
, Ahmet Çakır
5
1
Ağrı İbrahim Çeçen University, Eleşkirt Celal Oruç School of Animal Production, Animal Health Department. Ağrı, Türkiye.
2
Çukurova University, Faculty of Medicine, Department of Pathology. Adana, Türkiye.
3
Kirşehir Ahi Evran University, Faculty of Medicine, Department of Microbiology. Kırşehir, Türkiye.
4
Ağrı İbrahim Çeçen University, Faculty of Pharmacy. Ağrı, Türkiye.
5
Kilis 7 Aralık University, Faculty of Science and Literature, Department of Chemistry. Kilis, Türkiye.
*Corresponding author: syakan@gmail.com
Eects of Helichrysum plicatum DC. subsp. plicatum extract in a model of experimentally excited urolithiasis / Yakan et al.______
2 of 7 3 of 7
INTRODUCTION
Urolithiasis (U) is a disease caused by the development of stones
within the urinary system. About 80 percent of these stones consist of
calcium oxalate (CaOx). Besides CaOx stones, other types can result
from infections, uric acid, cystine, and calcium phosphate [1, 2, 3, 4,
5]. Subjecting renal cells to oxalate (Ox) and/or CaOx crystals results
in the formation of free oxygen radicals, leading to oxidative stress
(OS) and consequent damage and suppuration. Kidney damage and
inflammation are key factors in the formation of stones [6, 7, 8, 9, 10,
11]. Researchers have shown that treatments involving antioxidants
and free radical sweepers can decrease the harm caused by Ox/
CaOx crystals. Furthermore, the aggregation of CaOx crystals in the
kidneys is largely reduced by treatments with antioxidants and free
radical sweepers, providing evidence for the efcacy of antioxidants
[11, 12, 13, 14, 15]. Developing treatments to prevent OS could be an
alternative method that may raise the achievement ratio in avoiding
and intercepting future occurrences of kidney stones [12]. Since OS
occurs due to various sources and lanes, it is important to determine
all enzymes and lanes involved in the formation of free radicals caused
by Ox and/or CaOx crystals [6]. It can be argued that flavonoids can
play an effective role in protecting against many illnesses that may
occur as a result of damage caused by free radicals [16, 17].
Helichrysum species, which are rich in flavonoids, are used in
traditional medicine worldwide. This plant, which encompasses
hundreds of species, has been utilized in folk medicine for treating
wounds, and infections, and as a diuretic since ancient times. Türkiye
alone has about 20 varieties of Helichrysum, which are used in Turkish
traditional medicine for treating wounds, and burns, reducing kidney
stones, as diuretics, and for relieving ear pain. Previous research
has revealed that this plant possesses antimicrobial, antioxidant,
anti–inflammatory, antidiabetic, and anti–mitotic properties [18,
19, 20, 21, 22, 23, 24]. The medicinal features of Helichrysum
are believed to be attributed to flavonoids and other metabolites.
In a previous study, β–sitosterol, apigenin, nonacosanoic acid,
astragalin, β–sitosterol-3-O–β–D–glucopyranoside, helichrysin
A, helichrysin B, and isosalipurposide were isolated from the
HP. It has been observed that secondary metabolites extracted
from HP methanol have potent inhibitory effects, particularly on
human carbonic anhydrase I (hCAI) and II (hCAII) [24]. Although
Helichrysum species are extensively used in Turkish traditional
medicine, the lack of scientic evidence regarding their specic
clinical effects, activity, and side effects has prevented the clear
denition of their potential use as an alternative or complement
to conventional treatment. Therefore, further scientic research
is necessary to dene the exact mechanism of the effect of these
traditional remedies [12].
In this study, the antioxidant and anti–inflammatory action of
Helichrysum plicatum DC. subsp. plicatum (HP) methanol extract
was investigated in the urinary tract using an experimentally
excited urolithiasis model.
MATERIALS AND METHODS
Animals
Twenty–eight male Wistar albino rats (Rattus norvegicus) weight
140 ± 10 g and aged 10–12 weeks were used in the research. The
animals were housed in lattices in a controlled room at a constant
temperature of 24 ± 1°C and were provided with a twelve hour
(12 h) dark/light cycle. Prior to the study, the rats were consent to
acclimate to the perimeter for 10 days (d) without any interventions.
The Wistar albino rats were given access to feed (pellets) and water
ad libitum. The research was applied at Çukurova University Health
Sciences Experimental Research and Application Center (SABİDAM)
following ethical guidelines for experimental animal studies, and
conrmed by the Çukurova University Experimental Animals Ethics
Committee (conrmation no: 2019/75).
Experimental protocols
Twenty–eight male rats intended for use in the experiment were
divided into four equal groups. To induce the formation of CaOx
crystals, a mixture of 1% ethylene glycol (EG) and 1% ammonium
chloride (AC) was added to the swig water of the animals for 21 d.
The control and research groups were designed in the
following manner:
○ Group 1 (control, n:7): In rats in the control group, 1 mL of
water was given by gastric gavage method for 21 d.
○ Group 2 (HP only, n:7): The rats in this group received 500
mg·kg
-1
·day
-1
of HP via gavage feeding each morning for 21
d, along with their regular daily nutrients, under the same
laboratory conditions.
○
Group 3 (urolithiasis only, n:7): In the urolithiasis group,
1% EG and 1% AC were additional to the rats’ swig water
daily for 21 d to stimulate urolithiasis.
○
Group 4 (urolithiasis–HP, n:7): The animals in this group
were administered 500 mg·kg
-1
·day
-1
of HP via gavage
feeding each morning for 21 d, in addition to their daily
nutrients. They were also given 1% EG and 1% AC in their
swig water. The doses of HP and the urolithiasis model were
chosen based on previous studies.
In the experiment, all animals were anesthetized with ether using
a mask with an inhalation anesthesia device (Harvard Apparatus,
Anesthesia Machine, USA). Once they reached deep anesthesia, and
then immolate by taking approximately 3 mL blood samples from
their hearts for biochemical analysis. After the rats had died, their
abdominal cavities were immediately opened for histopathological
examination and the kidneys were quickly removed.
Biochemical measurements
Blood samples taken from rat hearts were centrifuged (Eppendorf,
5424R, Germany) at 3000 G for 10 min to obtain serum, which was
then preserved at -80 degrees until analyzed. The samples taken
from -80°C on the study day were thawed at room temperature.
High–performance Liquid Chromatography (HPLC) analysis
Measurement of HP in serum was conducted utilizing high–
performance liquid chromatography (HPLC) (Agilent Technologies,
1260 Innity II, USA). The analysis of HP serum concentration
by HPLC was carried out according to the methods outlined in
the literature [25, 26]. According to this, a C30 phase column
Eects of Helichrysum plicatum DC. subsp. plicatum extract in a model of experimentally excited urolithiasis / Yakan et al.______
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separation system was used to isolate HP in serum. 1 mL of serum
was homogenized in a 3 mL salt/ethanol mix (1:2, weight: volume).
Then, 5 mL of a hexane/ether mixture (1:1, volume: volume) was
incorporated, and the mix was vortexed for 3 min before being
centrifuged at 4°C at a speed of 2000 G for 10 min. The upper layer
was accumulated, and HP was separated from the samples. The
samples were extracted three times and treated with 100 mL of an
ethanol/ether mixture (2:1, volume: volume) and then evaporated
under nitrogen gas. The concentration of HP was measured by
injecting 50 mL of the nal sample into the instrument.
Determination of antioxidant and anti–inflammatory activity
Blood samples were taken on 9
th
days in group 1 and on 1,
9, 17, and 21 d in the other groups. Plasma concentrations of
thiobarbituric acid reactive substances (TBARS) were measurement
with spectrophotometer (Thermo Fisher Scientic, NanoDrop
2000, USA). The change in TBARS aimed to identify the antioxidant
and anti–inflammatory.
After the rats were sedated with ether, 1 mL of blood was taken
from their tails. Plasma TBARS levels, which are composed of two
molecules of Thiobarbituric acid (TBA) and malondialdehyde (MDA)
and serve as an indicator of oxidative membrane damage, were
determined following the fluorometric method described in the
literature [27, 28]. A solution of 29 mmol·L
-1
TBA (Sigma, T5500),
5 mol·L
-1
hydrochloric acid (HCI) (Sigma, H–7020), and n–butanol
(prepared in 8.75 M Acetic acid (Sigma, A-6283)) was used to
measure plasma TBARS levels. Standard solutions of MDA bis
(dimethyl acetal) (Sigma, T–1642) were prepared at concentrations
of 0; 0.5; 1; 2.5; 3.5; 5; 7; 8; 9; 10 µmol·L
-1
.
The plasma samples taken in anticoagulant tubes were
centrifuged at 3000 G for 10 min, aliquoted, and stored at -80°C
(Thermo Fisher Scientific, Forma 900 Series, USA) up to the
study day. On the day of the study, samples were thawed at room
temperature. For testing, 10 mL tubes each containing 1 mL of
distilled water were prepared for each blood specimen and 50 µl
of plasma was added. Then, 1 mL of the 29 mmol·L
-1
TBA solution
prepared in acetic acid was added to each tube and mixed. The
tubes were tightly closed and placed in a 100°C boiling water
bath for 1 h. After cooling the tubes under tap water, the pH was
adjusted to 1.6–1.7 by adding 25 µL of 5 mol·L
-1
HCl solution to
each tube. 3.5 mL of n–butanol was supplemented to the reaction
concoction, which was vortexed for 5 min.
Subsequently, the fluorescences of the upper phase divided by
centrifugation at 1500 G for 10 min were read using a fluorometer
(Thermo Fisher Scientic, Fluoroskan FL, Finland) at wavelengths
of 525 nm (excitation) and 547 nm (emission). The MDA contents
of the blood samples were calculated in µmol·L
-1
based on the
standard solutions.
Assays in blood and urine
Serum parameters: Oxalate levels were measured using the Trinity
Biotech diagnostic kit with spectrophotometer (Thermo Fisher
Scientic, NanoDrop 2000, USA). Concentrations of creatinine
and urea in the serum were determined using autoanalyzer
diagnostic (Siemens Healthineers, ADVIA 1800, Germany) kits with
spectrophotometer (Thermo Fisher Scientic, NanoDrop 2000, USA).
Urine parameters: On the 20
th
day of the experiment, two rats
from each group were placed in metabolic cages for 24 h to collect
their urine. The total urine volume was measured with a measuring
cylinder and recorded in milliliters. Ox levels were determined using
the Trinity Biotech diagnostic kit with a spectrophotometer (Thermo
Fisher Scientific, NanoDrop 2000, USA). Levels of creatinine
(measured with Jaffe’s method) and calcium (measured with the
Schwarzenbach and o–Cresolphthalein method) were determined
in the urine samples using an autoanalyzer (Siemens Healthineers,
ADVIA 1800, Germany). Creatinine clearance was calculated
according to the methods described in the literature [29].
Histopathological evaluation
The kidney tissues were xed in 10% formaldehyde for 3 h after
the cassette process was completed and sample numbers were
assigned. Subsequently, overnight tissue detection and tracking
were carried out on the fully automated device. Following tissue
xation, parafn embedding and blocking were performed. The
prepared parafn blocks were sampled with 5 μm sections. The
parts obtained were stained with hematoxylin–eosin using a fully
automatic staining device. The kidney tissue was examined under
a microscope (Olympus, BX53, Japan) for the presence of crystals
and inflammation. Crystals were scored based on their density
under a polarized lens. Mean group scores were determined based
on the scores of rats in the same group.
According to this scoring system: Score 0: no stones and clumps
of CaOx crystals; score 1: 1–5 stones, CaOx crystals; score 2: 5–10
stones, CaOx crystals; score 3: more than 10 stones, CaOx crystals.
Statistical analysis
The SPSS 22.0 package program was using for statistical analysis.
The data is adherence to a normal distribution was evaluated through
the Shapiro–Wilk test. Data that met the criteria for a normal
dispersion were analyzed usage analysis of variance and student
t–tests for group comparisons, as well as t–tests for dependent
variables for intra–group comparisons. Post–hoc comparisons were
conducted utilizing the Scheffe test. Data that did not meet the
criteria for a normal distribution were compared using Kruskal–Wallis
variance analysis and the Mann–Whitney U test.
RESULTS AND DISCUSSIONS
Biochemical ndings
Serum HP values
After administering HP to rats for 21 d, the average ± SD HP
value was found to be 340.5 ± 66.4 nmol·L
-1
in the HP group and
120.3 ± 19 nmol·L
-1
in the U–HP group. These values were found to
be statistically signicantly higher (P<0.001) compared to the group
1 (< 0.000 + 0) and U (< 0.000 + 0) groups where no HP was given.
When comparing the groups that received HP (HP, U–HP) with those
that did not (Control, U), the HP concentration in the HP groups was
statistically higher (P<0.001), indicating successful administration
via. Furthermore, when comparing the HP and U–HP groups, the HP
concentration was found to be signicantly lower in the U–HP group
(P<0.001). This difference was attributed to increased antioxidant
need in the U–HP group, leading to lower HP levels.
Eects of Helichrysum plicatum DC. subsp. plicatum extract in a model of experimentally excited urolithiasis / Yakan et al.______
4 of 7 5 of 7
TABLE I
The average of plasma TBARS changes in groups
*
TBARS (nmol·L
-1
)
Days Group 1 (Control) Group 2 (HP) Group 3 (U) Group 4 (U+HP)
1 – 0.026 ± 0.020 – 0.026 ± 0.020
9 0.026 ± 0.020 0.025 ± 0.010 0.021 ± 0.010 0.022 ± 0.001
17 – 0.020 ± 0.002 0.180 ± 0.240 0.025 ± 0.002
21 – 0.024 ± 0.001 0.121± 0.003 0.013± 0.003
*: Abbreviations are as follows; TBARS: thiobarbituric acid reactive substance, HP:
Helichrysum plicatum, U: urolithiasis
TABLE II
Average values and standard deviation (X
̄
± SD) of blood
and urine, stone formation score by groups
1
Parameters
Group 1
(Control)
Group 2
(HP)
Group 3
(U)
Group 4
(U–HP)
Serum oxalate
(µmol·L
-1
)
30.16 ± 15.24
a
34.79 ± 9.08
a
74.86 ± 60.81
a
36.76 ± 8.04
a
Serum urea
(mg·dL
-1
)
40.35 ± 3.27
b
40.14 ± 13.08
b
84.78 ± 60.34
a
* 41.36 ± 15.06
b
Urine oxalate
(µmol·24 h
-1
)
2.67 ± 0.46
c
2.65 ± 0.28
c
10.11 ± 5.12
a
** 7.04 ± 3.01
b
*
Urine calcium
(mg·dL
-1
)
6.60 ± 3.11
b
7.48 ± 4.31
b
12.89 ± 4.26
a
* 7.66 ± 2.13
b
Creatinine clearance
(mL·min
-1
)
0.22 ± 0.06
b
0.26 ± 0.16
b
0.55 ± 0.27
a
* 0.29 ±0.13
b
Stone formation
score
0.00 ± 0.00
c
0.00 ± 0.00
c
2.56 ± 0.48
a
** 0.21 ± 0.69
b
*
1
: Abbreviations are as follows; HP: Helichrysum plicatum, U: urolithiasis.
a,b,c
: There is
a statistically signicant dierence between values with dierent letters in the same
row (*
P<0.05, **P<0.001)
FIGURE 1. Photomicrography of kidney divisions illustrates varying group scores
based on stone density and calcium oxalate deposits. Score 0: Absence of stones
and deposits; Score 1: 1-5 stones with calcium oxalate crystal deposits; Score 2:
5-10 stones with calcium oxalate crystal deposits; Score 3: Over 10 stones with
calcium oxalate crystal deposits
Score 0
Score 2
Score 3
Score 1, polarized
Score 3, polarized
Score 2, polarizedScore 2, polarized
Plasma TBARS levels
Plasma levels of TBARS decreased signicantly between 9 and
17 d in the HP group (P=0.01). There was an important reduction
in TBARS concentrations between 17 and 21 d in the U–HP group
(P=0.003). TBARS levels increased signicantly from 9 to 17 d in
the U group (P=0.004). The TBARS level was signicantly riser
compared to the initial level (P=0.006) on 21 d in the U group.
On 17 d, TBARS levels were under in the HP group compared to
the U–HP group (P=0.044). Additionally, the TBARS level was
signicantly lower on 17 d (P=0.001) and 21 d (P=0.001) in the
U–HP group compared to the U group. The average TBARS levels
for all rats can be found in TABLE I.
Assays in blood and urine
The serum oxalate concentration in the U–HP group was higher
than in the control group but lower than in the U group, although
this increase was not statistically signicant. Even though the
serum Ox level was riser in group U than in other groups, there
was no important distinction in serum Ox levels between the groups
(P>0.05). The serum urea level was found to be signicantly higher in
the U group compared to the other groups in the group comparisons
(P<0.05). Urine Ox concentrations were signicantly riser in the U
group compared to the other groups (P<0.001). In the U–HP group,
urine Ox level was showed to be signicantly lower compared to the
U group (P<0.05). Urine calcium concentrations were signicantly
higher in the U group compared to the control, HP, and U–HP groups
(P<0.05). There was no important difference in the comparisons
between the control, HP, and U–HP groups (P>0.05). Creatinine
clearance was signicantly riser in the U group compared to other
groups (P<0.05). There was no important distinction in creatinine
clearance levels between the control, HP, and U–HP groups (P>0.05).
Biochemical measurement outcomes of blood and urine samples
are provided based on the groups in TABLE II.
Histopathological results
For histopathological examination, the stone creation density
of kidney tissues was scored between 0 and 3 after examining
all kidney sections. Mean group scores were then determined
(see TABLE II). In FIG. 1, photomicrographs of kidney sections
demonstrate diverse group scores with varying kidney stone
intensity. No abnormal histological formations were found in
the control and HP groups during the histological examination of
kidney tissue sections. However, the U group animals showed large
quantities of CaOx crystals deposited in masses. Stone creation and
accumulation were as expected in group U, with a stone creation
score of 2.56. The healing effect of HP extract on stone creation
was evident, as the stone creation score of the U–HP group was
lower (0.21) compared to the U group (2.56). Within the interstices
Eects of Helichrysum plicatum DC. subsp. plicatum extract in a model of experimentally excited urolithiasis / Yakan et al.______
_________________________________________________________________________________________________Revista Cientica, FCV-LUZ / Vol.XXXV
5 of 7
deposited mononuclear inflammatory cells were usually present
in the U group, less so in the U–HP group. Additionally, capsular
inflammatory lesions were detected in the U group.
Previous researches have demonstrated that Helichrysum
extracts are benecial for treating urolithiasis, a condition long
recognized in traditional medicine [30, 31]. This research aims
to contribute to a better understanding of the mechanism of the
HP effect in future studies.
Oxalate is a natural by–yield of metabolism and is excreted without
causing harm in healthy individuals. However, hyperoxaluria, which
is characterized by improved urinary excretion of oxalate, can lead
to crystal formation at normal pH levels and may result in the
accumulation of calcium oxalate crystals in the kidneys. Oxalate has
a stimulating effect on renal epithelial cells at low concentrations,
but at high concentrations, it can be toxic by causing the creation of
calcium oxalate crystals. The collection of calcium oxalate crystals
in the kidneys can trigger the production of certain macromolecules
that can cause the formation of pus and lead to brosis. Studies
on animals and renal epithelial cell cultures have indicated that
hyperoxaluria and the formation of calcium oxalate crystals can
lead to the generation of free radicals [32, 33, 34, 35, 36, 37, 38].
In a study made by Huang et al. [39], it was found that levels
of alpha glutathione S transferase, MDA, and TBARS in the
urine of CaOx kidney stone patients were elevated, indicating
oxidative stress in the kidneys. The study also revealed increased
urinary excretions of beta–galactosidase and N–acetyl–beta
glucosaminidase (NAG), which are indicators of renal epithelial
injury. Tungsanga et al. [40] observed in their studies that stone
patients exhibited higher levels of oxidative stress and renal tubular
cell injury. Urolithiasis patients were also found to have higher
plasma MDA, urinary MDA, and urinary NAG activity compared to
normal controls. Furthermore, decreased levels of glutathione and
cellular glutathione peroxidase activity, protein thiol, and vitamin
E were reported in urolithiasis patients. A study investigating the
oxidant and antioxidant status of blood in urolithiasis patients
before and after surgery reported a reduce in oxidant enzymes
and an rise in antioxidant status following surgical treatment
[41]. In another study, plasma TBARS levels were used to assess
membrane damage caused by oxygen radicals in renal tissue
affected by urolithiasis. The study suggested that higher plasma
TBARS levels in the group receiving HP (a treatment) compared
to the non–HP group may be due to the expected antioxidant
and anti–inflammatory properties of HP. While there was a slight
increase in plasma TBARS levels in HP patients, this increase
was not statistically signicant. The ndings indicated that HP
supplementation improved oxidative stress and reduced lipid
peroxidation products, supporting the antioxidant activity of HP
in the treatment of kidney stones.
In this study, biochemical parameters were evaluated the level
of creatinine clearance was found to be an important riser in the U
group according to the other groups due to damage to the kidneys.
The decrease in calcium ratios in the U–HP group according to the
U group in 24-h urine shows that hypercalciuria decreased with
the treatment we applied. In the amount of oxalate studied in the
urine, a very high amount of Ox was found in the U group compared
to the other groups. We found that HP suppressed urinary Ox
levels in the U–HP group. These ndings can be said to be one of
the preventive effects of HP stone creation.
With this study, observed that the concentration of creatinine
clearance was signicantly higher in the U group compared to the
other groups, due to kidney damage. The decrease in calcium ratios
in the U–HP group, compared to the U group, in 24-h urine samples,
indicates that hypercalciuria decreased with the treatment we
administered. We observed a higher amount of Oxalate in the urine
of the U group compared to the other groups. However, we found that
HP reduced urinary Oxalate levels in the U–HP group. These ndings
indicate HP may have a preventive effect on kidney stone creation.
CONCLUSIONS
In conclusion, we support the hypothesis that HP prevents the
formation of stones by correcting acute kidney damage through
its antioxidant and anti–inflammatory effects, thus preventing
oxidative stress.
Financial support
This study was not nancially supported by any institution
or organization.
Conflict of interest statement
The authors state that they have no conflicts of interest.
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