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EFFECTIVENESS OF RUTAN IN CORRECTING DISORDERS OF LIPID
PEROXIDATION IN PREPUBERTAL RATS WITH ACUTE TOXIC HEPATITIS
Ziyaviddin Z. Khakimov¹, Alisher H. Rakhmanov², Fatima A. Khalmuratova³
¹Doctor of Medical Sciences, Professor, Department of Pharmacology, Tashkent State
Medical University
²Doctor of Medical Sciences, Professor, Researcher, Biomedical Center, Tashkent State
Medical University
³Assistant, Department of Pharmacology and Pharmaceutical Technology, Karakalpak
Medical Institute, Uzbekistan
Abstract:
Considering the increasing incidence of toxic liver damage—the primary organ
for xenobiotic detoxification—the insufficient effectiveness of currently used
pharmacological treatments necessitates the search for and implementation of new effective
drugs for treating hepatobiliary system pathology in prepubertal patients. In this context, the
authors studied the effects of Rutan and Karsil on the levels of lipid peroxidation products
(Acylhydroperoxides [AcGP], malondialdehyde [MDA]) and the activity of antioxidant
enzyme systems (catalase [CAT], superoxide dismutase [SOD]) in the liver during acute
hepatitis induced by carbon tetrachloride in one-month-old rats. Results showed that Rutan,
similarly to Karsil, reduces elevated levels of AcGP and MDA in the microsomal-cytosolic
liver fraction significantly increased by acute toxic hepatitis induced by carbon tetrachloride.
Notably, this effect is accompanied by increased activity of antioxidant enzyme systems. It
was concluded that Rutan may be used in the treatment of acute liver diseases in the
prepubertal period.
Keywords:
acute toxic hepatitis, rats, prepubertal period, liver, free radical oxidation,
antioxidant defense enzymes.
Conflict of interest.
The authors declare no actual or potential conflicts of interest related to
the publication of this article.
Introduction
The widespread prevalence and high incidence of hepatitis present a serious medico-social
problem for healthcare systems. The use of many effective hepatoprotectors—derivatives of
amino acids and other organic acids, vitamin preparations, and compounds from various
chemical groups—is widely discussed globally [1]. In the development of toxic liver
pathology, free radical processes play a significant role [2]. One of the current problems in
hepatology remains the development and implementation of new highly effective
hepatoprotectors, as toxic liver disease incidence has increased in recent years [3,4]. At the
same time, the use of known therapeutic agents does not always yield the desired results. It
is believed that improving the effectiveness of pharmacotherapy for hepatobiliary system
pathologies requires a comprehensive approach using safe multifunctional hepatoprotectors
that protect hepatocytes from damage and restore their function. The most promising in this
regard is the use of phytopreparations, characterized by a broad spectrum of
pharmacological activity, high effectiveness at early stages of diseases, during slow-
progressing and chronic diseases, as well as in remission and rehabilitation periods [1,5,6].
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Furthermore, hepatoprotective agents of plant origin used in complex pharmacotherapy
potentiate the effects of basic medications and correct metabolic disturbances in the
damaged organ [7,8]. This problem is particularly relevant in pediatric practice, as the
incidence of acute hepatobiliary diseases in children tends to increase [9]. Previously, we
demonstrated the effectiveness of Rutan—an interferon inducer—in treating acute and
chronic inflammation in experimental models [10,11]. Considering that liver damage of
toxic etiology involves aseptic inflammation in its pathogenesis, it was hypothesized that
Rutan might exert hepatoprotective effects. Studies in adult animals confirmed the
hepatoprotective properties of Rutan [12]. However, the potential use of Rutan as a
treatment for hepatitis in prepubertal subjects remained unexplored. It was also assumed that
the antioxidant properties of Rutan’s polyphenols underlie its beneficial effects [13].
Aim of the Study
To evaluate the effectiveness of Rutan compared to Karsil on the intensity of lipid
peroxidation processes and the activity of antioxidant enzymes in acute toxic hepatitis in the
prepubertal period.
Materials and Methods
The study was conducted on 30 white growing rats of both sexes, one month old, born under
vivarium conditions. The experiments were carried out in accordance with the rules adopted
by the International Convention for the Protection of Vertebrate Animals used for
Experimental and other Scientific Purposes (Strasbourg, 1986). The conduct of experimental
research on laboratory animals was approved by the Ethics Committee of the Tashkent
Medical Academy under the Ministry of Health of the Republic of Uzbekistan (protocol No.
9 dated May 26, 2025).
Acute toxic hepatitis (ATH) was induced in the rats by intragastric administration of a 50%
oil solution of carbon tetrachloride (CCl₄) at a dose of 0.2 ml/100 g once daily for 4 days.
Twenty-four hours after the last administration of CCl₄, the animals were divided into four
groups. Rats in the first and second groups received Rutan at doses of 25 mg/kg and 50
mg/kg, respectively; the third group received Karsil at a dose of 40 mg/kg; and the fourth
group was left untreated (control relative to all other groups). The drugs were administered
once daily for six days.
The effects of Rutan and Karsil on the intensity of lipid peroxidation (LPO) processes and
the activity of antioxidant enzyme systems (AOS) in rats were studied. Twenty-four hours
after the last procedure, under light ether anesthesia, the rats were sacrificed in a cold room
at a temperature of 0–2°C by instantaneous decapitation. The liver was homogenized in a
glass homogenizer with a Teflon pestle in an isolation medium consisting of 0.25 M sucrose,
0.05 M KCl in a 0.05 M Tris-HCl buffer solution, pH 7.4.
To precipitate nuclei, mitochondria, and particles of damaged cells, the homogenates were
centrifuged at 9000 g for 20 minutes. In the microsomal-cytosolic fraction of the liver, the
content of acylhydroperoxides (AcGP) was determined by the method of Gavrilov V.V. and
Mishkorudnaya M.M., and the content of malondialdehyde (MDA) by the method of
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Andreeva L.I. [14]. The state of antioxidant enzyme systems (AOS) was assessed by the
activity of catalase (CAT) [14] and superoxide dismutase (SOD) [14].
The obtained data were statistically processed using the standard software package Statistica
for Windows by the well-known method of variation statistics with evaluation of
significance of the indicators (M±m) and differences between samples by Student’s t-test.
Differences between groups were considered significant at a 95% confidence level (p <
0.05).
Results and Discussion
It is well known that the formation of free radicals, which have a destructive effect on the
structure of biological membranes of cells, including hepatocytes, is one of the important
pathogenetic mechanisms of the damaging action of hepatotoxins [15–17]. This especially
applies to carbon tetrachloride [15–17]. The latter is widely used in experimental hepatology
in the study and introduction of new hepatoprotectors. Many researchers use the
determination of initial products — acylhydroperoxides (AcGP) and intermediate products
— malondialdehyde (MDA) of lipid peroxidation to assess the intensity of free radical
processes [6,14,18].
In this regard, we investigated the content of AcGP and MDA in prepubertal animals with
acute toxic hepatitis treated with Rutan and Karsil. The results of biochemical studies
showed a significant increase in LPO products in animals with acute toxic hepatitis.
Compared to intact animals, the AcGP content increased more than 4.5 times, and MDA —
3.7 times. These results confirm the high prooxidant activity of carbon tetrachloride [19,20].
In contrast, administration of Rutan at a dose of 25 mg/kg resulted in a decrease of AcGP
levels by 68.6% and MDA by 64.0% compared to untreated animals. Doubling the dose of
the drug did not lead to a noticeable change in this effect. Experimental pharmacotherapy
with Karsil, as seen from the data in Table 1, had a similar effect, i.e., the drug significantly
reduced the levels of AcGP and MDA to a degree not significantly different from the results
of Rutan.
Table 1.
The effect of Rutan and Karsil on the intensity of lipid peroxidation processes in the
liver of rats with acute toxic hepatitis, (M±m, n=6).
Animal groups
Doses,
mg/kg
Acyl hydroperoxide
rel. units / mg protein
Malondialdehyde nmol/mg
protein
Intact
-
0,75 ± 0,04
0,69 ± 0,06
OTG+H
2
O
-
3,41 ± 0,23
а
2,56 ± 0,28
а
OTG+Rutan
25
1,07 ± 0,08
а,б
0,92 ± 0,09
б
OTG+Rutan
50
1,28 ± 0,12
а,б
1,17 ± 0,11
а,б
OTG+Karsil
40
1,19 ± 0,13
а,б
1,08 ± 0,09
а,б
Note: a – P <0.05 compared to the intact group, b – P <0.05 compared to the control group.
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Therefore, the results of the conducted studies indicate that in acute toxic hepatitis (ATH),
there is a significant and persistent increase in the content of lipid peroxidation (LPO)
products in the microsomal-cytosolic fraction of the liver. Experimental therapy with Rutan
and Karsil led to a substantial decrease in the levels of Acylhydroperoxides (AcGP) and
Malondialdehyde (MDA), indicating suppression of LPO intensity.
It is known that the content of lipid peroxidation products depends on the activity of
antioxidant enzyme systems (AOS). Taking this into account, we studied the effect of Rutan
and Karsil on the activity of antioxidant enzymes in the liver of animals with hepatitis.
The results showed that six days after the induction of the hepatitis model, the activities of
catalase (CAT) and superoxide dismutase (SOD) decreased by 64.9% and 60.7%,
respectively, compared to the values of healthy animals (Table 2). In the experimental
therapy with Rutan at doses of 25 and 50 mg/kg in rats with ATH, a statistically significant
increase in CAT activity was observed at 131.9% and 97.9%, and SOD activity at 122.1%
and 103.2%, respectively, compared to untreated animals. A similar increase in the activity
of the studied enzymes was observed with Karsil therapy, with CAT activity increasing by
140.4% and SOD by 111.6% compared to untreated animals.
It should be noted that with treatment by both drugs, the activity of the studied enzymes did
not differ statistically significantly from the corresponding values of healthy animals.
Table 2.
Effect of Rutan and Karsil on the activity of antioxidant enzyme systems in the liver of
rats with acute toxic hepatitis (M±m, n=6).
Animal groups
Doses,
mg/kg
Catalase, nmol
H
2
O
2
/min ∙ mg protein
Superoxide
dismutase
conventional units/min ∙ mg
protein
Intact
-
1,34 ± 0,12
2,42 ± 0,14
OTG+H
2
O
-
0,47 ± 0,04
а
0,95 ± 0,04
а
OTG+Rutan
25
1,09 ± 0,11
б
2,11 ± 0,19
б
OTG+Rutan
50
0,93 ± 0,19
б
1,93 ± 0,19
б
OTG+Karsil
40
1,13 ± 0,10
б
2,01 ± 0,16
б
Note: a – P <0.05 compared to the intact group, b – P <0.05 compared to the control group.
Analysis of the results studying the effects of Rutan and Karsil on the activity of antioxidant
enzyme systems (AOS) allows concluding that the tested drugs restore the enzymatic
activity of catalase (CAT) and superoxide dismutase (SOD), resulting in a clear reduction of
lipid peroxidation (LPO) intensity. The obtained results correspond with data from other
researchers [10,11]. At the same time, the pharmacological efficacy of Rutan does not
significantly differ from that of Karsil.
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Thus, the results of experimental studies on the efficacy of Rutan in correcting disorders of
free radical lipid oxidation in prepubertal rats with acute toxic hepatitis (ATH) show that the
studied polyphenol-containing compounds suppress the intensity of free radical lipid
peroxidation in biological membranes by increasing the activity of AOS enzymes,
confirming the hepatoprotective activity of these compounds. Therefore, like Karsil, Rutan
can be recommended as a pathogenetic treatment option for toxic liver injuries of various
etiologies in pediatric practice.
Conclusions
1.
In prepubertal animals, ATH is accompanied by a significant enhancement of LPO
processes in hepatocyte biological membranes, which is manifested by an increased content
of LPO products in the microsomal-cytosolic liver fraction.
2.
The increased intensity of free radical lipid oxidation in ATH in growing animals is
accompanied by a decrease in the activity of AOS enzymes in liver cells.
3.
Experimental therapy of ATH with Rutan more effectively reduces the intensity of
free radical lipid oxidation in the liver compared to Karsil.
4.
The positive influence on the activity of antioxidant enzymes allows recommending
Rutan for complex treatment of many pathological conditions and diseases in whose
pathogenesis enhanced free radical oxidation during the prepubertal period may be a
contributing factor.
References:
1.
Blinova E.V., Khalzova M.A., Blinov D.S., Yakhya M.Kh.S., Meleshkin A.I.,
Kurganov N.A., Geroeva O.I. Hepatoprotective activity of an organic magnesium compound
in pregnant rats with experimental hepatitis. Experimental and Clinical Pharmacology. 2018;
81(4):18-21.
2.
Simonova N.V., Dorovskikh V.A., Bondarenko D.A., Nosal L.A., Shtarberg M.A.
Comparative efficacy of Remaxol and Reamberin in liver damage by carbon tetrachloride in
experiment. Experimental and Clinical Pharmacology. 2018; 81(7):29-33.
3.
Molochkova O.V., Kovalev O.B., Konev V.A., Uchaykin V.F., Snetkova Y.S. Drug-
induced hepatitis in children. Pediatric Infections. 2017; 16(1):42-50.
4.
Molostvova A.F., Khabirova G.I., Kharisova Yu.I., Salimova L.M. Drug-induced
liver damage - modern view on the topical problem. Bulletin of Modern Clinical Medicine.
2022; 15(5):107-115.
5.
Lupanova I.A., Ferubko E.V., Kurmanova E.N., Shishkanov D.V., Saibel O.L. Study
of hepatoprotective activity of dry extract of Jerusalem artichoke herb in a toxic hepatitis
model in rats. Pathological Physiology and Experimental Therapy. 2022; 66(4):138-142.
6.
Hakimov Z.Z., Rakhmanov A.Kh., Mavlanov Sh.R. Effectiveness of medicinal plant
extract mixtures in correcting liver functional status disorders of various etiologies. Tashkent,
TMA Publishing, 2021. – 156 p.
7.
Demidov V.N., Nazarenko O.A., Egorova E.Yu. et al. Efficacy of Progepar in
experimental liver damage caused by alcohol and paracetamol. Pharmateka, 2011; 2: 85–90.
8.
Nikolaev S.M. Phyto-pharmacotherapy and phytoprophylaxis of diseases. Ulan-Ude:
2012; 286 p.
Vo
lu
m
e
5,
Ju
ly
,2
02
5
,
M
ED
IC
AL
SC
IE
N
CE
S.
IM
PA
CT
FA
CT
OR
:7
,8
9
9.
Sema Mandal, Ruth Simmons, Georgina Ireland, et al. Pediatric acute hepatitis of
unknown etiology: a national investigation and adenoviraemia case-control study in the UK.
The Lancet Child & Adolescent Health. 2023; 7(11):786-796.
10.
Abzalov Sh.R., Khakimov Z.Z., Rakhmanov A.Kh. The Effect of Dry Extract
Obtained from Rhus Cariaria in Experimental Models of Edema and Inflammation.
American Journal of Medicine and Medical Sciences. 2020; 10(7):503-508.
11.
Khakimov Z.Z., Rakhmanov A.Kh., Abzalov Sh.R. Influence of RHUS Coriaria L.
leaf extract on chronic arthritis progression. Medical Journal of Uzbekistan. 2024; 2:182-188.
12.
Mavlonov A.A., Boboeva R.R. Study of the hepatoprotective action of Rutan.
Academicia: an International Multidisciplinary Research Journal. 2020; 10(5):117-120.
13.
Aripov T.F., Ghaibov U.G., Ghaibova S.N., Abdullaev A.A., Abduazimova D.Sh.,
Oschepkova Yu.I., Salikhov Sh.I. Antioxidant and antiradical activity in vitro of polyphenol
mixture (active substance of antiviral drug Rutan) from RHUS CORIARIA L. leaves.
Chemistry of Plant Raw Materials. 2024; 4:138-147.
14.
Mavlanov Sh.R., Khakimov Z.Z., Rakhmanov A.Kh. Experimental methods for
studying the effects of new pharmacologically active compounds on hepatobiliary system
function. Methodological Guide, Tashkent, 2017. – 63 p.
15.
Rakitskii V.N., Skupnevskii S.V. Seasonal variations of carbon tetrachloride toxicity.
Hygiene and Sanitation. 2019; 98(3):324-327.
16.
Timasheva G.V., Repina E.F., Karimov D.O., et al. Peculiarities of metabolic
changes in experimental animals at early terms of pharmacological correction of toxic liver
diseases. Toxicological Review. 2020;(1):13-19.
17.
Hsu Y-W, Chen Y-Y, Tsai C-F. Protective Effects of Chlorogenic Acid against
Carbon Tetrachloride-Induced Hepatotoxicity in Mice. Processes. 2022; 10(1):31.
18.
Gildikov D.I. Oxidative stress in animals: a pathophysiologist's perspective. Russian
Veterinary Journal. 2020; 4:10–18.
19.
Averyanova E.V., Shkolnikova M.N., Chugunova O.V., Mazko O.N. Effects of
triterpenoids in fatty products on liver condition of laboratory animals with acute toxic
hepatitis. Problems of Nutrition. 2023; 92(4):81-91.
20.
Unsal V., Cicek M., Sabancilar İ. Toxicity of carbon tetrachloride, free radicals and
role of antioxidants. Rev Environ Health. 2020 Sep 25;36(2):279-295.
