INTRODUCTION:

T. indica L, widely known as tamarind, is a tropical arboreal species originating from Africa and Indonesia.

It is also used as a cooking spice and an alternative source of nutrition for food. Its culinary and medicinal properties have led to widespread cultivation in various regions globally¹. Tamarind has been used for centuries in Ayurvedic and traditional restorative practices to address different health conditions, including cardiovascular disorders. According to recent research, tamarind contains various bioactive compounds that exhibit potential health benefits such as antioxidant, anti-inflammatory, and lipid-lowering properties².

Increased total cholesterol concentrations in the circulatory system substantially threaten the onset of cardiovascular ailments, such as atherosclerosis and coronary artery disease. The abovementioned conditions are the primary contributors to illness and death globally ³. The management and mitigation of overall cholesterol levels are pivotal in averting and impeding the initiation and advancement of such ailments. Although there are conventional medications for reducing lipids, they frequently produce unfavorable side effects. Consequently, many researchers are interested in investigating natural alternatives, specifically medicinal plants, to regulate cholesterol levels⁴.

The research on the biological impact of T. indica in regulating total cholesterol is significant due to its potential to offer a natural and secure method for controlling cholesterol levels⁵. The potential ability of the bioactive compounds found in tamarind to modulate total cholesterol levels in the human body may present a viable alternative or supplementary approach to traditional pharmacological interventions⁶.

The antioxidant properties of T. indica have been acknowledged due to diverse bioactive constituents such as polyphenols, flavonoids, and vitamin C⁷. As mentioned, the compounds exhibit scavenging properties towards free radicals, conferring cellular protection against oxidative stress and reactive oxygen species-induced damage⁸. Research has indicated that tamarind possesses antioxidant properties that could potentially aid in preventing hypercholesterolemia by impeding the oxidation of LDL cholesterol and diminishing oxidative stress within the body⁹. Tamarind can potentially mitigate atherosclerosis and cardiovascular ailments linked to elevated cholesterol levels by minimizing oxidative harm. Additionally, there have been reports indicating that tamarind possesses properties that can lower lipid levels, potentially playing a role in preventing hypercholesterolemia. Several research studies have suggested that tamarind extract or its bioactive constituents can decrease total cholesterol, LDL cholesterol, and triglyceride levels in animal models and in vitro experiments. These effects may be attributed to bioactive compounds, such as polyphenols and flavonoids, which exhibit antioxidant and lipid-lowering properties¹⁰.

It is imperative to comprehend the mechanisms underlying the impact of tamarind on cholesterol metabolism to authenticate its customary application and potentially create innovative therapeutic interventions. Examining tamarind's biological effect on the regulation of total cholesterol in a preclinical model, specifically Mus musculus (more commonly referred to as mice), can provide significant knowledge regarding its potential therapeutic uses in the human population. The present study aims to significantly contribute to developing natural remedies based on empirical evidence. These remedies are intended for individuals who prefer non-pharmacological methods to regulate their cholesterol levels. Additionally, if the efficacy of tamarind in decreasing overall cholesterol levels is established, it may have broader ramifications for the prevention and management of cardiovascular ailments ¹¹.

In general, investigating the biological impacts of T. indica on the regulation of total cholesterol in Mus musculus represents a vital research undertaking that could yield valuable findings regarding the possible utilization of tamarind as a natural therapeutic intervention. The results obtained from this investigation have the potential to facilitate additional scholarly inquiry, such as experimental studies, to substantiate its effectiveness and safety in human subjects, ultimately providing novel approaches for treating disorders related to cholesterol and enhancing cardiovascular well-being. This study aimed to test and analyze the potential of T. indica to prevent or reduce the increase in total cholesterol in the blood of the Mus musculus and reduce the degeneration and infiltration of fat in the tissues in the liver and coronary arteries.

Material and Methods:

This research has passed ethical approval for animal model No. Ref 009/KEPH-C/VII/2017 from the Faculty of Veterinary Medicine, Universitas Syiah Kuala, Darussalam, Banda Aceh, Indonesia. This study used mice (Mus musculus) and ethanol extract of T. indica as the active material to test its biological properties in reducing total cholesterol in Mus musculus.

Assay Material:

This study used six treatment groups, each consisting of 4 mice, as follows: Group (1) Negative group, normal mice; (2) Positive group, hypercholesterolemia; 3) Anti-cholesterol drugs, Simvastatin 10 mg BW/day; (4) T. indica 35 mg; 5) T. indica 175 mg; 6) T. indica 350 mg. Treatment of atherogenic (hypercholesterol) animal models using high-fat feed with control thresholds. Groups label: G0 (A control treatment showing normal coronary artery histology); G1 (Represents a hypercholesterolemia treatment showing more fat and changes in the shape of the coronary arteries), G2 (Shows normal coronary artery histology); G3 (Shows the amount of fat reduced when compared to G1); G4 (Shows the amount of fat is reduced when compared to P3); G5 (Shows normal coronary artery histology)

Plant Material:

Tamarandus indica was obtained from Ie Masen Village, Ulee Kareng District, Banda Aceh City, Aceh Province, Indonesia, coordinates point 5.55550N 95.3560E. Taxonomic identification was carried out in the Biology Laboratory of the Teaching and Education Faculty of Syiah Kuala University, Banda Aceh, Indonesia. Extracting and storing T. indica is carried out and stored in the Chemistry Laboratory of the Faculty of Teaching and Education Faculty, Syiah Kuala University, Banda Aceh, Indonesia. The storage of the test material refers to the laboratory sample storage protocol.

Extract Preparation:

The manufacture of ethanol extract of T. indica was adopted, starting with the separation of the skin and the separation of the flesh and seeds. Furthermore, at room temperature, as much as 1kg of meat is dried for three days and then mashed and mashed using a blender. At room temperature, as much as 50g of T. indica extract was immersed in 250mL of 96% ethanol. Then, it is stirred for 1 hour and stored in a dark place for 24hours. The extract is filtered using filter paper and evaporated to separate the solvent. Evaporation was carried out using a rotary evaporator with a speed of 3,000rpm at 57°C for 10 minutes until the solvent evaporated utterly to obtain a thick tamarind fruit extract. The prepared extract was stored at -20°C until use¹².

DPPH antioxidant assay:

The investigation of C. odorata's antioxidant properties was conducted utilizing the DPPH (2,2-diphenyl-1-picryl-hydrazine-hydrate) principle, as Salazar-Aranda et al. (2011) outlined. A solution was prepared by dissolving 100mg of the sample and vitamin C in 1 mL of DMSO, followed by vortexing. Furthermore, 100 µL of the sample and vitamin C were incorporated into the 96-microplate. Subsequently, 100µL of DPPH solution was introduced, while the negative control was supplemented with 100µL of ethanol. Later, the sample was incubated at ambient temperature for 30 minutes. Subsequently, the model was subjected to spectrophotometric analysis at a specific wavelength of 517nm¹³.

Hypecholesterol of Animal Model:

Twenty-four Mus musculus males, two months old, weighing approx. 20g were acclimatized at 28±2°C) to regular light/dark cycles and free access to food and water for one week before use. After acclimatization, animals were randomly separated into six groups of four rats. Five groups were given a high-fat diet of 60% corn flour, 8% fish meal, 20% soybean meal, 3% egg yolk, 6% coconut oil, 1% premix, 1% salt, and 1% CaCO₃. A high-fat diet is prepared by mixing all the ingredients thoroughly and baking in the oven at 40°C overnight¹⁴. Feed is made in the form of pellets before being given to animals. Another group, recorded as standard control (NC), was fed normal rats. Water and diet were provided ad libitum for thirty days. The experimental protocol and animal handling during the study complied with the guidelines approved by the institution's ethical committee where the research was conducted¹⁵.

Cholesterol Assay:

In this experiment, treatment with T. indica was initiated after 30 days of atherogenic feed induction and designated as day 30¹⁶. Three doses of T. indica were used: 35mg/20g BW, 175mg/20g BW, and 350mg/20g BW. Supplementation was administered daily by oral gavage for 30 days, with a volume of 0.2mL/20g body weight. The negative control (NC) and positive control (PC) groups were given distilled water as a placebo. Total blood cholesterol concentration was measured twice at the beginning (before treatment) and at the end. Blood was collected after 12 hours of fasting from the lateral tail vein of the mice by dripping blood from the tail vein and waiting for the total cholesterol result to appear on the screen. Total cholesterol was examined using Autocheck (Autochek Easy Touch, Manufacture Bioptic Technology, Germany). The assessment of triglyceride, LDL, and HDL levels in plasma was analyzed using the cholesterol oxidase-p-aminophenozone (CHOD-PAP) reagent, triglycerides using the glycerol phosphate oxidase-p-aminophenozone (GPO-PAP) test reagent, and glucose with the glucose oxidase-test reagent, p-aminophenozone (GOD-PAP) (Boehringer Mannheim Kit from Germany).

Histopathology Assessment:

The retrieval of animal model tissues or organs began with euthanasia using an intravenous injection of ketamine at 0.2mg/kg BW. Organs or tissues were stored in formalin buffer preservatives and prepared for pathological examination. The liver and heart were the pathological tissues/organs examined. The preparation process included dehydration with 70% to absolute alcohol, clearing in xylol, and infiltration and embedding in paraffin blocks at 56-58°C. The embedded tissues were sliced to a thickness of 5 microns using a rotary microtome. For histopathological analysis, liver and heart specimens fixed in 10% neutral buffered formalin were embedded in paraffin, sliced 5mm thick, and stained with H&E stain to detect hepatic and cardiac steatosis. Pathological changes were assessed and photographed under an Olympus U-CMAD3 microscope ¹⁷. Liver biopsy grading was: 0 (no steatosis, normal liver), I (25% of hepatocytes affected), II (26–50% of hepatocytes affected), III (51–75% of hepatocytes affected), and IV (76% of hepatocytes affected). Heart coronary artery biopsy followed the methodology of Walker (1992)¹⁸.

Statistical Analyses:

The data obtained were analyzed using the Statistical Package for the Social Sciences (SPSS) version 16 program. These data were analyzed using a paired T-test and a one-way analysis of variance (ANOVA). The p<0.05 value was used to indicate statistical significance.

Results:

Based on the IC₅₀ antioxidant examination results, T. indica has an antioxidant content of 10.45 ppm. In comparison, the control vitamin C contains 9.79 ppm. In principle, the fewer active ingredients needed to reduce DPPH, the stronger the antioxidant quality. Table 1 data is representative of Figure 1. 7 chemical compounds have the dominant content in Tamarindus L-pulp, including the highest peaks No. 16 (5-Hydroxymethylfurfural, 28.66%), 21 (Hexadecanoic acid (CAS) Palmitic acid, 16.84%), 19 (Beta.-D-Glucopyranose, 1,6-anhydro-, 11.08%), 22 (Oleic Acid, 7.96%), 4 (2,3-Dimethylenebutane-1,4 -diacetate, 5.82%), 20 (1,6-Anhydro-beta-D-glucopyranose, 5.16%), and 3 (2-Furancarboxaldehyde (CAS) Furfural, 3.39%)

Table 2 reported that the subject animal model (Mus musculus), which was given atherogenic feed, showed increased cholesterol (pre-treatment). The group treated with T. indica L showed a decrease within the normal range for 30 days. The standard feed group decreased by 3%, and the atherogenic feed group increased by 29% (simvastatin decreased by 35%, (T. indica 35 mg decreased by 16%, T. indica 175 mg decreased by 36%), and T. indica 350 mg decreased by 40%. The standard value used in this study is the normal limit category of 80-130 mg/dL ¹⁹.

Table 1. Chemical compounds of T. indica and its quantity

Peak #	Compound Name	Rate Time	Spectrum Area	Area %
1	Formic acid	5.558	42565455	0.64
2	Acetic acid	5.748	56413109	0.84
3	2-Furancarboxaldehyde (C.A.S.) Furfural	5.945	227411959	3.39
4	2,3-Dimethylenebutane-1,4-diacetate	6.097	389753235	5.82
5	2(5H)-Furanone	8.350	97096376	1.45
6	2,4-Dihydroxy-2,5-dimethyl-3(2H)-furan-3	8.856	27446831	0.41
7	2H-Pyran-2,6(3H)-dione (C.A.S.) Glutaconic	9.187	16259754	0.24
8	2,5-Furandione, dihydro-3-methyl-	9.555	12816621	0.19
9	Benzeneacetaldehyde (C.A.S.) Hyacinthin	9.700	19144489	0.29
10	Methyl 2-furoate	10.616	187796045	2.80
11	2-Furanmethanol (C.A.S.) Furfuryl alcohol	11.012	131105457	1.96
12	2-Butene, 1,4-diethoxy-	11.200	40301073	0.60
13	4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy	11.824	253800668	3.79
14	2-Ethyl-4-hydroxy-5-methyl-3(2H)-furanone	12.430	26273035	0.39
15	5-Acetoxymethyl-2-furaldehyde	12.666	43776565	0.65
16	5-Hydroxymethylfurfural	14.180	1920061889	28.66
17	3-Heptanol (C.A.S.) 3-Hydroxyheptane	14.903	126987124	1.90
18	2-Propenoic acid, 3-phenyl-	16.005	87833882	1.31
19	.beta.-D-Glucopyranose, 1,6-anhydro-	18.568	742277788	11.08
20	1,6-Anhydro-beta-D-glucopyranose	19.662	345465490	5.16
21	Hexadecanoic acid (CAS) Palmitic acid	21.511	1128072197	16.84
22	Oleic Acid	23.266	533492951	7.96
23	2-Cyclohexene-1-carboxylic acid, 1-methyl	26.236	176153811	2.63
24	E,E,Z-1,3,12-Nonadecatriene-5,14-diol	30.731	43974545	0.66
25	Oxirane, hexadecyl-	31.155	22405513	0.33

Table 2. Profile of total cholesterol of Mus musculus after treatment by T. indica

Groups	N	*Effect of T.Indica L* on the total cholesterol**
		Pre-Treatment	*p-value	Treatment	*p-value	Total Cholesterol
		Mean ± SD	*p-value	Mean ± SD	*p-value	Freq (%)	Status
Standard feed	4	105,25 ± 2,217	0.001	101,50 ± 1,00	0.001	-3%	Decrease
Atherogenic feed	4	136,25 ± 13,07	0.001	150,50 ±15,02	0.001	29%	Increase
Simvastatin 10 mg	4	130,75 ± 7,54	0.001	103,00 ± 2,16	0.001	-35%	Decrease
T.indica 35 mg	4	135,25 ± 12,81	0.001	123,00 ± 4,96	0.001	-16%	Decrease
T.indica 175 mg	4	142,00 ± 23,50	0.001	114,00 ± 2,44	0.001	-36%	Decrease
T.indica 350 mg	4	134,00 ± 6,37	0.001	102,75 ± 2,98	0.001	-40%	Decrease
**p-value	24	0.421		0.041

* Paired T-test; ** One Way Anova

Table 3. Profile of HDL of Mus musculus after treatment by T. indica

Groups	N	Effect of T.Indica L on the HDL
		Pre-Treatment	*p-value	Treatment	*p-value	HDL
		Mean ± SD	*p-value	Mean ± SD	*p-value	Freq (%)	Status
Standard feed	4	83.45±11.63	0.001	83.45±6.52	0.001	1%	Increase
Atherogenic feed	4	50.87±1.51	0.001	45.98±8.58	0.001	-5%	Decrease
Simvastatin 10 mg	4	49.15±1.51	0.001	88.69±3.28	0.001	28%	Increase
T.indica 35 mg	4	47.30±1.25	0.001	83.05±2.94	0.001	24%	Increase
T.indica 175 mg	4	51.05±2.66	0.001	85.55±5.09	0.001	24%	Increase
T.indica 350 mg	4	50.46±3.79	0.001	88.62±2.02	0.001	28%	Increase
**p-value	24	0.0310		0.021

* Paired T-test; ** One Way Anova

Table 4. Profile of LDL of Mus musculus after treatment by T. indica

Groups	N	Effect of T.Indica L on the LDL
		Pre-Treatment	*p-value	Treatment	*p-value	LDL
		Mean ± SD	*p-value	Mean ± SD	*p-value	Freq (%)	Status
Standard feed	4	27.46±2.00	0.001	27.21±2.66	0.001	-0.2%	Decrease
Atherogenic feed	4	61.48±5.18	0.001	64.99±9.47	0.001	2%	Increase
Simvastatin 10 mg	4	64.29±5.18	0.001	49.78±4.91	0.001	-10%	Decrease
T.indica 35 mg	4	65.49±2.35	0.001	21.38±6.45	0.001	-30%	Decrease
T.indica 175 mg	4	65.07±2.77	0.001	19.40±3.73	0.001	-31%	Decrease
T.indica 350 mg	4	62.75±6.46	0.001	16.07±1.39	0.001	-32%	Decrease
**p-value	24	0.020		0.010

* Pairet T-test; ** One Way Anova

Table 5. Profile of triglycerides of Mus musculus after treatment by T. indica

Groups	N	Effect of T.Indica L on the Trygliserida
		Pre-Treatment	*p-value	Treatment	*p-value	Triglycerides
		Mean ± SD	*p-value	Mean ± SD	*p-value	Freq (%)	Status
Standard feed	4	92.52±9,32	0.001	95.03±6,52	0.001	-1.5%	Decrease
Atherogenic feed	4	173.52±6,42	0.001	173.27±6,42	0.001	0,2%	Increase
Simvastatin 10 mg	4	170.56±6,71	0.001	122.76±10,66	0.001	-29%	Decrease
T.indica 35 mg	4	167.42±24,87	0.001	155.13±9,75	0.001	-8%	Decrease
T.indica 175 mg	4	172.91±17,62	0.001	131.41±8,56	0.001	-26%	Decrease
T.indica 350 mg	4	170.14±17,20	0.001	106.97±4,46	0.001	-39%	Decrease
**p-value	24	0.036		0.041

* Pairet T-test; ** One Way Anova

Figure 2. Histopathological Micrograph Photo of Coronary Arteries of the Heart after Induction of Atherogenic and Antihypercholesterol Feeding. (a). Tunica intima, (b). Tunika Media, (c) Tunika Adventisia, (d). Lumens (e). Fat infiltration, (f) Fat degeneration. (Magnification 400x). G0 (A control treatment showing normal coronary artery histology); G1 (Represents a hypercholesterolemia treatment showing more fat and changes in the shape of the coronary arteries), G2 (Shows normal coronary artery histology); G3 (Shows the amount of fat reduced when compared to G1); G4 (Shows the amount of fat is reduced when compared to P3); G5 (Shows normal coronary artery histology)

Discussio:

This study has evaluated the potential of T. indica to prevent hypercholesterolemia by lowering total cholesterol, LDL, triglycerides, and lower HDL. This potential implies that the antioxidant compounds possessed by T. indica have a role in preventing the formation of excessive fat in the blood. Antioxidant levels of T. indica tested in this study ranged from 10.45 ppm IC50, with vitamin C containing 9.79 ppm as a control. An indication that antioxidants work to neutralize hypercholesterolemia is the occurrence of decreased degeneration and fatty infiltration in the liver and coronary arteries of the heart. Several antioxidant compounds that T. indica L has are in very high quantities, among other compounds. Antioxidant compounds are essential in lysing blood fats to trigger cholesterol anemia ²¹.

Trapani (2011) reported that one potential theoretical mechanism by which antioxidants in plants can help block hypercholesterolemia is their ability to mitigate oxidative stress and modulate lipid metabolism. Hypercholesterolemia is associated with increased reactive oxygen species (ROS) levels and oxidative stress ²². Oxidative stress occurs when there is an imbalance between the production of ROS and the body's antioxidant defense mechanisms. Hypercholesterolemia refers to high blood cholesterol levels, mainly low-density lipoprotein (LDL) cholesterol ²³. This condition is a significant risk factor for developing cardiovascular diseases, including atherosclerosis, characterized by forming plaques in the arterial walls ²³. Oxidative stress occurs when there is excessive production of ROS, such as superoxide anions, hydrogen peroxide, and hydroxyl radicals, or a reduced capacity of the body's antioxidant defense mechanisms to neutralize these harmful molecules ²⁴. ROS are natural byproducts of cellular metabolism and involve various physiological processes. However, when their production exceeds the antioxidant capacity, they can lead to oxidative damage to cells, proteins, lipids, and DNA ²⁵.

Tables 2, 3, 4, and 5 reported that T. indica can reduce total cholesterol, LDL, and triglycerides and increase blood HDL. This phenomenon indicates that several antioxidant compounds in T. indica ethanol extract, apart from having solid binding affinities with cellular proteins, can also independently work to inhibit cholesterol synthesis by inhibiting the genes involved in the formation of LDL. The antioxidant compounds in T. indica extract may exert their cholesterol-lowering effects through various mechanisms. Firstly, these compounds can directly interfere with the expression and activity of key enzymes involved in cholesterol synthesis, such as 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase). By inhibiting the function of HMG-CoA reductase, the rate-limiting enzyme in cholesterol biosynthesis, the antioxidant compounds in T. indica can effectively reduce the production of cholesterol, including LDL cholesterol ²⁶. Furthermore, the antioxidant compounds in T. indica extract may modulate gene expression related to cholesterol metabolism. By targeting specific genes involved in LDL formation, these compounds can downregulate their expression, thereby inhibiting the synthesis of LDL particles ²⁷.

The ability of T. indica to increase HDL cholesterol, often referred to as "good cholesterol," further supports its potential in improving lipid profiles. The antioxidant compounds in T. indica may enhance the expression and activity of genes involved in HDL metabolism, promoting the transport and removal of cholesterol from peripheral tissues to the liver for further processing and excretion ²⁸. It's important to note that while the specific antioxidant compounds in T. indica responsible for these effects may vary, the overall findings suggest that T. indica extract possesses cholesterol-lowering properties through multiple mechanisms, including direct inhibition of cholesterol synthesis and modulation of gene expression related to cholesterol metabolism ²⁹.

Table 6 and Figure 1 show that T. indica improves infiltration and prevents fatty degeneration in liver tissue in animal models of hypercholesterolemia. This potential indicates that several anti-inflammatory and antioxidant properties in T. indica prevent fat receptors from producing excess fat in the liver. Tamarindus indica has shown promise in improving liver health by reducing infiltration and preventing fatty degeneration in animal models of hypercholesterolemia. This beneficial effect suggests the presence of multiple anti-inflammatory and antioxidant properties in T. indica that work together to avoid excessive fat production in the liver ³⁰.

One potential mechanism behind the liver-protective effects of T. indica is its ability to attenuate inflammation ³¹. Chronic inflammation is closely associated with the development of fatty liver disease, including hypercholesterolemia-induced hepatic steatosis. The anti-inflammatory properties of T. indica can help suppress the production of proinflammatory cytokines and reduce the activation of inflammatory pathways, thereby mitigating liver inflammation and subsequent fat accumulation ³². Moreover, the antioxidant properties of T. indica contribute to its protective effects on liver tissue. Oxidative stress plays a significant role in the progression of liver diseases, including hypercholesterolemia-induced fatty liver ³³. The antioxidant compounds in T. indica can scavenge reactive oxygen species (ROS) and reduce oxidative stress, thus preventing cellular damage and lipid peroxidation in the liver. By maintaining a balance between antioxidants and ROS, T. indica helps protect liver cells from oxidative damage and fatty degeneration ³⁴.

In addition to its anti-inflammatory and antioxidant properties, T. indica may modulate lipid metabolism in the liver, preventing excess fat production ³⁵. The bioactive compounds in T. indica may regulate key genes and enzymes involved in lipid synthesis, uptake, and metabolism. By influencing these pathways, T. indica can inhibit the excessive fat accumulation in liver tissue, reducing the risk of hepatic steatosis. These mechanisms contribute to the protective effects of T. indica on liver health and its potential as a therapeutic agent for liver diseases associated with Hypercholesterolemia ³⁶.

Table 7 and Figure 2 show that T. indica improves infiltration and prevents fatty degeneration in the coronary artery. This potential indicates that several anti-inflammatory and antioxidant properties in T. indica avoid the formation of fatty boluses in the coronary artery or inhibit plaque formation. Besides that, it increases blood flow and oxygen supply through oxidative phosphorylation in the heart. It has demonstrated the potential to improve coronary artery health by reducing infiltration and preventing fatty degeneration ³⁷. This beneficial effect suggests the presence of multiple anti-inflammatory and antioxidant properties in T. indica, which work together to inhibit the formation of fatty deposits or plaques in the coronary artery. T. indica may enhance blood flow and oxygen supply to the heart by promoting oxidative phosphorylation. One potential mechanism behind the protective effects of T. indica on the coronary artery involves its anti-inflammatory properties. It plays a critical role in developing atherosclerosis, which consists of the buildup of fatty deposits in the arterial walls ³⁸.

The anti-inflammatory compounds in T. indica can help mitigate inflammation by suppressing the production of proinflammatory cytokines and inhibiting the activation of inflammatory pathways. By reducing inflammation, T. indica may prevent the accumulation of fatty deposits and impede the progression of atherosclerosis in the coronary artery ³⁹. The antioxidant properties of T. indica contribute to its potential to avert fatty deposition in the coronary artery ⁴⁰. Oxidative stress is a critical factor in atherosclerosis development, as it promotes lipid peroxidation and the formation of oxidized lipids, leading to plaque formation. The antioxidants in T. indica can scavenge reactive oxygen species (ROS) and protect against oxidative damage. By reducing oxidative stress, T. indica helps maintain the integrity of the coronary artery and prevents the formation of fatty plaques ⁴¹.

T. indica may enhance blood flow and oxygen supply to the heart by promoting oxidative phosphorylation. Oxidative phosphorylation is the process by which cells generate energy through adenosine triphosphate (ATP) through oxygen utilization ⁴². By improving oxidative phosphorylation, T. indica may enhance the heart's energy production, increasing blood flow and oxygen delivery to the coronary arteries. This enhanced oxygen supply can support the heart's metabolic needs and contribute to its overall health ⁴³. The combined anti-inflammatory, antioxidant, and cardiovascular effects of T. indica contribute to its potential to prevent fatty deposition and promote coronary artery health ⁴⁴. However, it's important to note that further research is needed to understand the specific mechanisms involved fully and to validate these theoretical propositions.

Conclusion:

Tamarindus indica L can reduce total cholesterol, LDL, and triglycerides and increase HDL in animal models of hypercholesterolemia. This ability aligns with the potential of antioxidant compounds and the bioactivity strength of T. indica compounds in neutralizing liver and heart function as indicated by decreased infiltration and degeneration of deep fat in the liver and coronary arteries microscopically.

Conflict of Interest:

The authors agreed there were no conflicts of interest.

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Received on 01.06.2024 Revised on 07.10.2024

Accepted on 04.01.2025 Published on 12.06.2025

Available online from June 14, 2025

Research J. Pharmacy and Technology. 2025;18(6):2756-2765.

DOI: 10.52711/0974-360X.2025.00395

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Treatment Groups	Histopathological Features	Pathological Status
G0	The histological features of the liver show that the liver's main elements are polyhedral and hexagonal hepatocytes with a round nucleus. Among the hepatocytes are sinusoids, which are capillary blood vessels that carry blood to the central vein.	Does not show pathological changes
G1	The histopathological features of the G1 group show a lot of fatty liver consisting of fat infiltration (fat vacuoles outside hepatocyte cells) and fatty degeneration (formation of fatty vacuoles that fill the cytoplasm of hepatocytes). Almost all parenchyma of the liver has fatty acids.	Indicates a change (shift) in the amount of fat
G2	Histopathologically, there was a significant reduction in infiltration and fat degeneration in this group. The hepatocytes appear standard with a central nucleus and regular sinusoids and resemble the G0 group.	It shows a minimal amount of fat degeneration and fat infiltration and resembles G0 (Normal) treatment.
G3	The G3 group saw a lot of fat infiltration and degeneration, but when compared to the P1 group, there was a decrease but not significant	shows the amount of fatty degeneration and fat infiltration is still very much
G4	The decrease in fat infiltration and degeneration was more significant in this group than in the G3 group. Hepatocyte and sinusoid cells began to appear regularly.	Shows less amount of fat and fat infiltration
G5	The reduction in fat infiltration and degeneration was significant compared to the G3 and G4 groups, where a marked decrease in fat amount was seen within and between hepatocyte cells.	Shows hepatocyte cells and sinusoids have started to be seen clearly

Groups Treatment	Histopathological Features	Pathological Status
G0	Histologically, the coronary artery group G0 looks intact (normal), consisting of the tunica intima, tunica media, and tunica advertising.	It shows no pathological changes, and there was no change in the thickness of the tunica intima. Endothelial cells appear intact.
G1	In the GI group, there was fatty tissue histopathologically marked by the presence of empty fat spaces (fat vacuoles) found on the surface of the tunica intima (fat infiltration) and the inside of the tunica media (degeneration). The artery turns slightly elliptical.	Changes occur in the coronary arteries, and there is a thickening of the tunica intima layer.
G2	Histopathological of coronary artery group G2 showed a typical feature: no fatty or atheroma plaque. The coronary arteries comprise the tunica intima, tunica media, and adventitia.	It showed a normal arterial picture; no fatty deposits or atheroma plaques were found.
G3	Histopathologically, the coronary arteries in group G3 showed fatty forms (fat infiltration) on the surface of the tunica intima, but the number was reduced compared to group G1.	Visible forms of fat (fat infiltration) on the surface of the tunica intima
G4	In the P4 group, the arterial shape was standard, and there was still fat in the form of fat infiltration, which was significantly reduced compared to G3 and G1.	The shape of the arteries returns to normal, although there is still fat in the form of significantly reduced fat infiltration.
G5	In the P5 group, the arterial shape was expected, and there was no fat either in the form of infiltration or degeneration.	There is no longer visible fat degeneration and fat infiltration in the tunica intima layer, so there is no longer any thickening.