International Journal of Virology and Molecular Biology

p-ISSN: 2163-2219    e-ISSN: 2163-2227

2025;  14(6): 156-164

doi:10.5923/j.ijvmb.20251406.13

Received: Oct. 7, 2025; Accepted: Nov. 3, 2025; Published: Dec. 30, 2025

 

Hepatoprotective Activity of Isoquinoline Alkaloids (F-5, F-24), and Their Derivatives (KV-6, KV-8) in CCl4-Induced Hepatotoxity

Abdubokiev R. Abduvali 1, Khushmatov S. Shunkor 2, Komilov J. Esoxon 3, Makhmudov Lazizbek Umarovich 4, Zhurakulov N. Sherzod 5

1Namangan State University, Republic of Uzbekistan

2Ministry of Higher Education, Science and Innovation of the Republic of Uzbekistan

3Institute of Biophysics and Biochemistry of National University of Uzbekistan, Tashkent, Republic of Uzbekistan

4Institute of Bioorganic Chemistry, Uzbek Academy of Sciences, Tashkent, Republic of Uzbekistan

5Institute of the Chemistry of Plant Substances, Uzbek Academy of Sciences, Tashkent, Republic of Uzbekistan

Correspondence to: Abdubokiev R. Abduvali , Namangan State University, Republic of Uzbekistan.

Email:

Copyright © 2025 The Author(s). Published by Scientific & Academic Publishing.

This work is licensed under the Creative Commons Attribution International License (CC BY).
http://creativecommons.org/licenses/by/4.0/

Abstract

Experiments have shown that under the conditions of experimental CCl4 toxic hepatitis, significant changes in the external morphoanatomical structure of the mouse liver are observed. In the experimental group, under the influence of isoquinoline alkaloids (F-5, F-24) and their flavonoid conjugates (KV-6, KV-8), positive dynamics were observed, the “liver index” values were optimized, and survival rates significantly increased.

Keywords: Isoquinoline alkaloids, CCl4-induced hepatotoxity, Hepatoprotective effect

Cite this paper: Abdubokiev R. Abduvali , Khushmatov S. Shunkor , Komilov J. Esoxon , Makhmudov Lazizbek Umarovich , Zhurakulov N. Sherzod , Hepatoprotective Activity of Isoquinoline Alkaloids (F-5, F-24), and Their Derivatives (KV-6, KV-8) in CCl4-Induced Hepatotoxity, International Journal of Virology and Molecular Biology, Vol. 14 No. 6, 2025, pp. 156-164. doi: 10.5923/j.ijvmb.20251406.13.

Article Outline

1. Introduction
2. Materials and Methods
    2.1. Morpho-Anatomical Analysis of the Liver in CCl4-Induced Hepatotoxity, and Method of Analyzing Mouse Survival
    2.2. Statistical Analysis
3. Results and Discussion
    3.1. Molecular Mechanism of Experimental Toxic Hepatitis Induced by CCl4
    3.2. The Influence of Isoquinoline Alkaloids (F-5, F-24) and Their Derivatives (KV-6, KV-8) on the Morphoanatomical Indices of the Liver in Experimental Toxic Hepatitis Caused by CCl4
    3.3. The Effect of Isoquinoline Alkaloids (F-5, F-24) and Their Derivatives (KV-6, KV-8) on the Survival of Mice with Experimental Toxic Hepatitis Caused by CCl4
4. Conclusions

1. Introduction

In the body, the liver plays an important role in normal physiological and pathophysiological processes of detoxification, carbohydrate, protein and lipid metabolism, and maintaining the functional homeostasis of the endocrine and immune systems. In turn, hepatoprotection is considered to be of great importance for the functioning of the human body as a whole and for its health [1,2,3,4,5].
It is noted that the number of liver diseases worldwide is dynamically increasing under the influence of various exogenous/endogenous factors, resulting in 2 million people dying from these diseases annually. In turn, the development of effective therapeutic approaches and drugs for hepatoprotection is of current importance [6,7,8,9,10].
It is also noted that many pharmacological drugs currently used in clinical practice have a negative, toxic effect on liver function, and therefore the creation of effective hepatoprotective drugs is relevant [11,12,13,14,15].
It has been confirmed that significant morphostructural changes in hepatocytes occur in hepatopathologies [16,17,18,19,20].
The aim of this study was to characterize the hepatoprotective activity of isoquinoline alkaloids (F-5, F-24) and their derivatives (KV-6, KV-8) in experimental toxic hepatitis caused by CCl4.

2. Materials and Methods

The experiments were conducted in 2023-2024 in the Pharmacological laboratory of the O.S.Sodikov Institute of Bioorganic Chemistry of the Academy of Sciences of the Republic of Uzbekistan. In the experiments, white mice (♀/♂, m=20.5-26.2 g) were used as research objects, receiving standard food (water) under standard vivarium conditions (room temperature t=+20±5°C, relative air humidity 75±10%, light regime 12:12 h) [21].
When working with experimental animals in scientific research, the requirements of the rules developed by the International Council for International Organizations of Medical Sciences (CIOMS) (1985), the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes (Strasbourg, 1986), the Declaration developed by the European Union (86/609/EEC) – Regulations on bioethics of the Institute of Biophysics and Biochemistry of the National University of Uzbekistan (No. BRC/IBB; N44/2024/75-1) were observed [22,23,24]. (Table 1)
Table 1. Chemical structure of isoquinoline alkaloids used in the studies – F-5, F-24, and their derivatives (KV-6, KV-8)
     
The objects of the research were isoquinoline alkaloids (F-5, F-24) and their conjugates with the flavonoid quercetin (KV-6, KV-8), synthesized by employees of the S. Yu. Yunusov Institute of Plant Chemistry of the Academy of Sciences of the Republic of Uzbekistan (Table 2) [25,26,27,28,29].
In studies, the hepatoprotective activity of isoquinoline alkaloids and their flavonoid conjugates was evaluated in a standard manner using a model of experimental toxic hepatitis in mice induced by CCl4 [30,31,32,33,34,35].
To create an experimental model of toxic hepatitis, we used white outbred mice of both sexes (♂♀, m=20.5-26.2 g) aged 2-3 months, kept in vivarium conditions on standard food and water. In the experiments, CCl4 solution (diluted in olive oil) was administered intraperitoneally (1 ml/kg) for 14 days (on days 1, 5, 11) [36,37,38,39,40,41,42].

2.1. Morpho-Anatomical Analysis of the Liver in CCl4-Induced Hepatotoxity, and Method of Analyzing Mouse Survival

The liver of experimental animals was preserved in formalin solution (10%) for morphoanatomical analysis [Mondal et al., 2022; pp.1013-1022] (Figure 1).
In experiments analyzing the hepatoprotective activity of isoquinoline alkaloids (F-5, F-24) and their derivatives (KV-6, KV-8) in CCl4-induced hepatotoxity, the survival rate (%) of mice was estimated using the standard method – the Kaplan-Meier method [43,44,45,46,47,48].
The survival analysis curve developed by Kaplan E.L., Meyer P. (1958) was calculated based on the following formula (2.1) [49,50,51,52,53,54,55]:
(2.1)
S(t) – is the survival rate at a given time (t), x – is the initial value of the survival function, (x+ti) – is the difference in values at the time of death, l(x+ti) – is the number of deaths that occurred before time x+ti, and dx+ti – is the number of deaths observed during time x+ti.
Figure 1. Experimental Design. To create an experimental model of toxic hepatitis, animals were administered CCl4 (1 ml/kg) on days 1, 5, and 11. In an experimental model of CCl4-toxic hepatitis, the hepatoprotective activity of isoquinoline alkaloids (F-5, F-24) and their derivatives (KV-6, KV-8) was analyzed by administering the substances to mice at a concentration of 100 mg/kg (orally mixed with olive oil (1:1)) on days 3, 6, 9, and 11 (the control group received an equivalent amount of olive oil to the experimental group) [Elshafei et al., 2013; pp.82-89]

2.2. Statistical Analysis

Mathematical and statistical processing of the obtained experimental results was carried out by standard methods using specialized software packages “Microsoft Excel 2007” (Microsoft, USA), OriginPro v. 8.5 SR1 (EULA, USA). The results are presented as M±m, where M is the arithmetic mean and m is the standard error of the mean, calculated based on the results of experiments conducted with n=3-4 repetitions. Also, the level of statistical significance of the values between the experimental results and the control group was calculated based on Student’s t-test and was considered statistically significant at values of p<0.05 and p<0.01. The level of reliability of the difference between the values of the two experimental groups was calculated using the Student’s t-test [56,57,58,59].

3. Results and Discussion

3.1. Molecular Mechanism of Experimental Toxic Hepatitis Induced by CCl4

Carbon tetrachloride (CCl4) is widely used to induce a model of toxic damage to hepatocytes in the form of experimental hepatitis in laboratory animals (in vivo), which is characterized by changes in ALT, AST, malondialdehyde, glutathione, superoxide dismutase, c-Jun N-kinase (JNK), glutathione peroxidase, catalase, tumor necrosis factor-α (TNF-α), alkaline phosphatase, bilirubin, total protein, albumins, quantitative and functional indices of globulins, hepatosis, fibrosis, cirrhosis, and hepatocellular degenerative changes [60,61,62,63].
CCl4-toxic hepatitis is an ideal experimental model for analyzing the hepatoprotective activity of various drugs in liver diseases in experimental animals [64,65,66,67].
Under the influence of CCl4, the induction of oxidative stress in hepatocytes with the participation of TNF-α (tumor necrosis factor-α), TGF-β1 (transforming growth factor-β1) and IL-1β factors leads to inflammation, collagen accumulation and fibrosis through the activation of a cascade of degradative inflammation and the accumulation of collagen and fibrin outside the hepatocyte membrane, which leads to fibrosis/cirrhosis (Fig. 2) [68,69,70] (Fig. 3).
Figure 2. Molecular mechanism of experimental induction of hepatitis by CCl4 [Ke and Lee, 2018; pp.214-223; Ma et al., 2022; pp.111-122; Bishnolia et al., 2025; pp. 1-8; Salim et al., 2025; pp.1-12]
Activation of nuclear factor erythroid-associated factor 2 (Nrf2), NF-κB, is believed to play an important role in the pathogenesis of experimental toxic hepatitis induced by CCl4 in hepatocytes [Wang et al., 2022; pp.1-14].
Exposure to CCl4 increases the concentration of cytokines (IL-1βa, IFN-γ) and chemokines (MCP-1, MIP-2β, KC), which are activated during inflammatory responses in mouse hepatocytes [71].
Moreover, experimental toxic hepatitis in rats induced by CCl4 was associated with a sharp increase in serum ALT, AST and bilirubin levels, as well as significant pathological changes in the morphoanatomical structure of the liver [72].
Structural and functional changes in hepatocytes under the influence of CCl4 have been described in detail by a number of researchers. It is emphasized that with the participation of cytochrome P450 (CYP2E1 and other isoforms), CCl4 forms free radicals – CCl3•, CCl3OO•, which in turn leads to the formation of lipid peroxidation based on a chain reaction of free radical generation, induction of apoptosis and necrosis [73].
Under the conditions of CCl4-toxic hepatitis, activation of the TNF-a/IL-6, TGF-b1/CTGF, lipid peroxidation (LPO) cascade, and collagen accumulation in hepatocytes (The involvement of MAPK/JNK – mitogen-activated protein kinase (Mitogen-activated protein kinase/c-Jun NH2-terminal kinase), Akt/NF-kB – transcription factor (Akt regulates the transcriptional activity of nuclear factor-kB) in the hepatotoxic process has also been confirmed), the development of fibrosis is noted [74,75].
Under conditions of experimental hepatitis induced by CCl4, a significant increase in liver weight, pronounced morphoanatomical deformation and histopathological changes were confirmed in experimental animals [76,77].

3.2. The Influence of Isoquinoline Alkaloids (F-5, F-24) and Their Derivatives (KV-6, KV-8) on the Morphoanatomical Indices of the Liver in Experimental Toxic Hepatitis Caused by CCl4

As a result of the experiments, it was established that under the conditions of CCl4-experimental toxic hepatitis, pronounced changes in the external morpho-anatomical structure of the liver of mice are observed, and in the experimental group, positive changes were observed under the influence of isoquinoline alkaloids (F-5, F-24), and their derivatives (KV-6, KV-8) (Fig. 3).
Figure 3. External morphoanatomical appearance of the liver of mice under healthy conditions (control) and toxic hepatitis CCl4 (The effect of the KV-6)
Typically, the average body weight of 3-4 month old mice is 20-35 g, the “liver index” is 0.060 (liver weight 1.89 g) in males and 0.058 (liver weight 1.27 g) in females. In experimental hepatitis caused by CCl4, animals exhibit a significant decrease in body weight, an enlarged liver and, in turn, an increase in the “liver index” value [78,79].
In the experiments, the average body weight in the control group was 23.40±0.37 g for males and 22.63±0.14 g for females, while under conditions of experimental toxic hepatitis, a decrease to 22.57±0.18 g and 21.15±0.09 g, respectively, was observed. In the physiological norm (control) state, the average liver weight of mice was m=1.05±0.01 g (mmin.=0.68±0.01 g, mmax.=1.46±0.01 g), and under experimental toxic hepatitis conditions it increased by 13.4±0.17% compared to the control (m=1.19±0.01 g, mmin.=0.94±0.01 g, mmax.=1.52±0.01 g). It was found that the liver index value averages 0.045 and 0.046 in males and females, respectively, and increases by 17.80% (0.053) and 21.74% (0.056) in toxic hepatitis compared to the control group, respectively (Table 2).
Table 2. The effect of isoquinoline alkaloids (F-5, F-24), their derivatives (KV-6, KV-8) on the liver index in mice with CCl4-experimental toxic hepatitis (M±m)
     
Under the influence of F-5, the liver mass was m=1.12±0.01 g, and the “liver index” value was 0.049 and 0.051 in male and female mice, respectively. The value of this indicator was recorded at the level of 0.050 and 0.052 for F-24 (100 mg/kg) (m=1.15±0.01 g) in male and female mice, respectively. Under the influence of KV-6 (100 mg/kg), the liver mass was m=1.08±0.01 g, and the “liver index” value was 0.046 and 0.049 in male and female mice, respectively. The value of this indicator for KV-8 (m=1.10±0.01 g) was 0.048 and 0.050, respectively.
The analysis of the obtained results showed that isoquinoline alkaloids (F-5, F-24) and their flavonoid conjugates (KV-6, KV-8) have significant hepatoprotective activity under conditions of experimental toxic hepatitis caused by CCl4, and this activity has a characteristic increase dynamic in the series F-24>F-5>KV-8>KV-6.

3.3. The Effect of Isoquinoline Alkaloids (F-5, F-24) and Their Derivatives (KV-6, KV-8) on the Survival of Mice with Experimental Toxic Hepatitis Caused by CCl4

During the experiments, it was established that in the experimental model of CCl4-toxic hepatitis, the S(t) value on the 4th, 7th and 14th days was 60.0, 46.7 and 33.3%, respectively, and significantly increased under the influence of isoquinoline alkaloids (F-5, F-24) and their derivatives (KV-6, KV-8) (Table 3).
Table 3. Analysis of survival rates of mice under conditions of CCl4-induced hepatotoxity
     
It was established that the value of this indicator was 73.3, 60.0 and 53.3% for isoquinoline alkaloids for F-5, respectively, 66.7, 66.7 and 46.7% for F-24, as well as 86.7, 80.0 and 73.3% for KV-6 for derivatives (F-5, F-24), respectively, 80.0, 66.7 and 60.0% for KV-8.
This experimental model is also a convenient method for analyzing the effects of various agents on the survival rates of experimental animals in studies using the Kaplan-Meier method [80,81].
In this regard, it has been confirmed that in an experimental model of hepatitis, toxic CCl4, the survival rate of mice is on average 70% [82,83].
The figure below shows the Kaplan-Meier estimate of the survival rate (%) of isoquinoline alkaloids (F-5, F-24), and their derivatives (KV-6, KV-8) in CCl4-induced hepatotoxity (Figure 4).
Figure 4. Kaplan-Meier survival estimate (%) of isoquinoline alkaloids (F-5, F-24), and their derivatives (KV-6, KV-8) in mice in CCl4-induced hepatotoxity. The median value reflects the death of 50% of animals by a given point in time (t)

4. Conclusions

Thus, in an experimental model of toxic CCl4 hepatitis, it was found that isoquinoline alkaloids (F-5, F-24), and their derivatives (KV-6, KV-8) significantly increase the survival rate of mice.

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