Cardioprotective Effect of Allium sativum Extract Against Doxorubicin Induced Cardiotoxicity in Rats

Shaguftha Sacline, Rasamalla Lavanya, Sontireddy Sai Krishna, Angirekula Mukesh, Puligilla Shankaraiah*

SVS Group of Institutions, Bheemaram, Hanamkonda, Telangan, India

*Correspondence: drspuligilla@gmail.com

DOI: https://doi.org/10.71431/IJRPAS.2026.5620  

INTRODUCTION

In addition to being used as a chemotherapeutic mediator for a wide range of hard cancers and malignancies, doxorubicin is an anti-cancer candidate drug that also causes myocardial damage. Doxorubicin's iron-anthracycline complex causes oxidative stress, which ruthlessly damages the membrane layer and interferes with the morphology of the cytoskeleton[1]. Doxorubicin in the iron-anthracycline complex causes oxidative stress and free radicals that target the cardiac tissue [2].

The development of atherosclerotic plaques is referred to as atherosclerosis, an inflammatory condition of the major arteries. Increased free radicals raised and modified low mass lipoprotein cholesterol, hypertension, diabetes mellitus, genetic changes, and significant plasma homocysteine absorption are all potential causes of endothelial dysfunction leading to atherosclerosis [3] . The most common cause of atherosclerosis-related clinical events, including MI or ischemic stroke, is the rupture of a susceptible atherosclerotic lesion [4]. Lipid peroxidation causes irreversible damage to myofibrils, cytoplasmic dialation with sarcoplasmic reticulum vacuolization, an increase in lysosomes, myocyte necrosis and inflammation of mitochondria [5], biosynthesis of protein and nucleic acid is also impeded[6], adrenergic alteration, Na+K+ATPase also shrinks [7], the cell used lysosomal and mitochondrial enzymes to convert doxorubicin into semiquinone. By voluntarily adding oxygen to an electron, the electric semiquinone creates oxidative stress, which releases free radicals. The conversion of superoxide to hydrogen hydroxide was mediated by superoxide dismutase [8].CVD is a collection of conditions affecting the heart and blood arteries, including atherothrombotic conditions like myocardial and cerebral infarction, which are referred to as heart attacks and strokes, respectively. the development of atherosclerotic plaque and endothelial dysfunction [9].

Garlic has protective effects against stroke, coronary thrombosis, atherosclerosis, platelet aggregation, as well as infections and vascular disorders. However, we must note that there is considerable controversy concerning the cholesterol lowering effects of garlic and a number of studies have reported that some garlic preparations do not lower serum cholesterol. The fibrinolytic activity of garlic in both man and experimental animals has been reported.

METHODOLOGY

Extraction procedure

The dried, finely powdered lower parts of Allium sativum become crude powder. Soxhlet was used to extract the crude powder using the solvent ethanol until it was colorless. After that, it was pressure evaporated using a rotary evaporator below 45ºC to wait for the extract's syrupy development before being moved to a clean dish at room temperature. The yield started at 21.5% w/v.

Phytochemical analysis

Ethanolic extract of Allium sativum  consists as Triterpenoids, flavonoids, tannins, saponins, glycosides, sterols, carbohydrates, alkaloids, and proteins were all qualitatively analyzed using the original syrupy extract [10].

Chemicals and animals

Thirty Wister albino rats of either sex, weighing an average of 202.11 ± 3.2 grams, were obtained for the study. The experimental rats were fed regular food and unlimited water for the duration of the study. Tests and animal husbandry were routinely conducted in accordance with ethical guidelines. In order to reduce undue stress, animals were acclimated to the laboratory environment for two days before to the study. The rats are kept in a 12-hour light-dark cycle at a standard laboratory temperature of 24±3ºC.Care was taken to avoid any tense provisions. The entire experiment was carried out after the Institutional Animal Ethical Committee approval (IEAC), provided the necessary materials for all of the tests, and doxorubicin were bought locally. The chemicals, the water and glassware used for this reading were of a systematic score and standard.

Experimental procedure

The following experimental design is used to evaluate the cardioprotective efficacy of Ethanolic Extract Allium sativum against Doxorubicin (DOX)-induced cardiotoxicity. Five groups of six rats each were randomly assigned to the rats.

Group I: Control Group: received water for 14 days.

Group II: DOX-induced group: received a single dose of DOX 15mg/kg I.P on 1st day only.

Group III: received DOX 15mg/kg I.P on the 1st day and 100mg EEAS for other 14 days.

Group IV: received DOX 15mg/kg I.P on the 1st day and 200mgEEAS for other 14 days.

Group V: a single dose of DOX 15mg/kg I.P on 1st day, Metoprolol (10 mg/kg bw) for the other 14 days.

After being anesthetized with ketamine and xylazine at the end of the fourteenth day, the animals were sacrificed with a large dose of ketamine. For enzyme tests, blood samples were collected right away. The blood samples were centrifuged for five minutes at 3000 rpm to create the serum. The rats' hearts were swiftly removed, cleaned in isotonic saline, homogenized in ice-cold 0.1M Tris HCl buffer (pH-7.5), and then stored.

Biochemical markers of the heart

Using standard kits, the LDH, AST, and CK-MB, SOD LPO and CAT activities were measured from serum in accordance with standard procedure[11].

Statistical analysis

The gathered data were expressed as the mean ± SD. Dunnett's test and ANOVA were used to balance the groups. P values less than 0.05 were considered significant.Steroids, tannins, flavonoids, phenolic compounds, carbohydrates, glycosides, alkaloids, saponins, and terpenoids were found through phytochemical analyses.

RESULTS

In the cardiac tissue, doxorubicin causes endogenous oxidative stress, which lowers SOD, CAT, and increases LPO. When compared to the control group, treatment with EEAS 200 and 300 mg/kg shows a significant rise (p<0.05) in SOD, CAT and significant decrease (p<0.05) in LPO. In the tissue next to oxidative stress, especially in cardiac tissue, SOD, CAT, and GSH are essential antioxidant biomolecules (Table 1).

Mammals in the DOX-induced group also exhibit muddy, yellowish to reddish, black hair development. The animals' exudates are black. The animals' poor health, sluggish movement, and swollen stomach and liver set them apart from the control group. When comparing the EEAS-treated group (100 & 200 mg/kg bw) to the DOX-induced animal group, the aforementioned annotations are clearly compressed. Rats given doxorubicin exhibit decreased body mass on the last day (Figures 1).When compared to the DOX-induced group, EEAS (100 mg/kg and 200 mg/kg) significantly increased body, heart, and its ratio (p<0.05)

The EEAS results for the CK-MB and AST enzymes were shown in (Table 2). Treatment among DOX sources with altitude in the quantity of oxidative enzymes, which might be computed as the cautious indicators for myocardial injury when compared to standard. This procedure demonstrated the reduction of the aforementioned oxidative enzymes in a greater quantity. Treatment with EEAS 100 mg/kg , EEAS 200 mg/kg showed a significant decrease in AST, LDH, and CK-MB (p<0.05). When compared to the control group, the protein content of the DOX-induced rats showed a decline. Compared to the DOX-induced group, animals administered with 100–200 mg/kg bw of EEAS showed significantly higher protein levels.

Figure 1: Body weight of Rats comparison on 1^st and 14^th  day in control, DOX-induced group, EEAS (100 mg/kg and 200 mg/kg) treatment Groups (*p<0.05,***p<0.001)

Table1: Antioxidants,CAT and LPO comparison on 14^th  day in control, DOX-induced group, EEAS (100 mg/kg and 200 mg/kg) treatment Groups (*p<0.05,***p<0.001)

Table-2: Cardiac Markers comparison on 14^th day in control, DOX-induced group, EEAS (100 mg/kg and 200 mg/kg) treatment Groups (*p<0.05,***p<0.001)

DISCUSSION

Through doxorubicin-induced cardiac toxicity, the ethanolic lower extract of Clitoria ternatea has revealed significant cardio protection in rats, a single administration of doxorubicin (15 mg/kg bw) will cause cardiotoxicity [12]. In this study, we created a cardiotoxic rat model by administering a single dosage of 15 mg/kg bw of DOX, which resulted in a significantly reduced body, heart, and mass ratio. This study's decrease in body mass is consistent with other research [13] and can be attributed to the animals' diet, which may reduce protein production because of doxorubicin. The ethanolic extract will show an increase in the body's mass, heart rate, and ratio. Creatinine kinase [14]  and LDH [15] both significantly increased as a result of DOX induction. With EEAS treatment, an increased level of these oxidative enzymes will return to normal significantly. The rats' illness is caused by DOX induction, which results in mitochondrial malfunction and oxidative stress. According to [16] Morin et al. (2001), DOX will produce reactive aldehyde, membrane lipid peroxidation, and ROS.When comparing the DOX-induced group to the normal control group, SOD and CAT enzymes showed a significant drop but an increase in LPO. EEAS treatment significantly restores these enzyme levels to normal in the DOX-induced group. This shows that EEAS prevents oxidative damage [17] When the control group received 100 mg/kg and 200 mg/kg bw of EEAS, the heart's architecture was nearly normal. Triterpenoids, flavonoids, tannins, saponins, glycosides, sterols, carbohydrates, alkaloids, proteins, and carbohydrates are examples of phytochemical constituents that strengthen the cardioprotective effects of EEAS in DOX-induced cardiotoxicity in the experimental rats.

CONCLUSIONS

When DOX induced cardiotoxicity and myocardial necrosis in the experimental animal, EEAS showed a convincing anti-cardiotoxic effect. LDH, CK-MB, and AST activities may be impacted by EEAS treatment. This suggested that EEAS may be the cause of the decrease and prevention of the serious cardiac side effects that doxorubicin causes. The EEAS extract is a unique strategy designed to prevent cardiotoxicity caused by doxorubicin. In conclusion, the overstatistics shown to support EEAS have the ability to act as an antioxidant to prevent the car-diotoxic effects of doxorubicin.

ACKNOWLEDGMENT

We would like to thank management of SVS Group of institutions, Hanamkonda,  for providing all the facilities to carry out my research work.

CONFLICT OF INTEREST STATEMENT

We declare no conflict of interest.

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