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. 2024 May 21;12(8):5776–5784. doi: 10.1002/fsn3.4229

Phytochemicals profiling of Cassia fistula fruit extract and its effect on serum lipids and hematological parameters in high‐fat diet‐induced hyperlipidemic female rats

Maryam Tariq 1, Nazir Ahmad 1,✉, Mahr Un Nisa 1, Muhammad Abdul Rahim 2,3,✉, Eliasse Zongo 4,✉
PMCID: PMC11317677  PMID: 39139970

Abstract

Cassia fistula (C. fistula) has shown strong anti‐inflammatory, hepatoprotective, antitussive, antibacterial, and antifungal properties and is being used for healing wounds and gastrointestinal illness. This study was planned to obtain fruit extract from C. fistula using ultrasonic‐assisted extraction (UAE) technique and evaluated for phytochemical contents, anti‐hyperlipidemia, and hematological parameters. The results showed that total phenolic (TPC), total flavonoids (TFC), condensed tannin (CT), and saponins were 13.07 mg GAE/g, 5.24 mg QE/g, 4.01 mg/g, and 27.55%, respectively, in the extract. Proximate composition of the extract showed 2.48%, moisture, 1.25% fat, 2.80% ash, 4.59% fiber, 11.93% protein, and 76.95% NFE. The 2, 2‐diphenyl‐1‐picrylhydrazyl (DPPH) and ferric reducing antioxidant power assay (FRAP) activity was 63.30 μg/mL and 15.02 nmol/g, respectively. High‐fat diet (HFD)‐induced hyperlipidemic rats were orally administrated with 0.5 and 1.0 g of extract/kg body weight (bw) daily. The reduction of total cholesterol (TC: 90.83 ± 8.86 mg/dL), triglycerides (TG; 74.16 ± 9.10 mg/dL), and low‐density lipoproteins (LDL; 74.83 ± 4.66 mg/dL) and increase of high‐density lipoproteins (HDL; 41.83 ± 8.4 mg/dL) was observed. Significant changes in red blood cells (RBCs; 8.03 ± 0.67106/μL), mean corpuscular hemoglobin concentration (MCHC; 35.02 ± 1.78 g/dL), mean corpuscular hemoglobin (MCH; 18.00 ± 0.26 pg), and mean corpuscular volume (MCV; 55.36 ± 4.01 fL) at 1.0 g extract intake was observed. Extract administration also improved significantly liver enzymes, body weight, and liver morphology. Therefore, C. fistula extract can be effectively used as a therapeutic agent to improve serum biochemistry and hematological values.

Keywords: C. fistula fruit, extract, hyperlipidemia, total phenolic contents, UAE

The purpose of the current research was to extract and evaluate the therapeutic potential of natural C. fistula pod extract against hyperlipidemia.

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1. INTRODUCTION

Hyperlipidemia is a metabolic disorder characterized by abnormal blood lipid levels, resulting in damage of endothelial bloody vessels (Fioranelli et al., 2018; Jaffe & Karumanchi, 2024; Rafaqat et al., 2024). This damage leads to a loss of nitric oxide in an affected region, ultimately escalating an inflammatory response and promoting lipid accumulation in an endothelial wall and leading to a formation of foam cells. Subsequently, activated macrophages engulf these foam cells and result in development of atherosclerosis, necrosis, mitochondrial dysfunction, and apoptosis (Cao et al., 2017; Howell et al., 2011; Naser et al., 2021). According to WHO, a global prevalence of hyperlipidemia is estimated to be 37% and 40% in males and females, respectively. Notably, 98.1% and 97.3% of newly diagnosed diabetics and pre‐diabetic individuals exhibited dyslipidemia, while prevalence in non‐diabetic Pakistani population is around 95.2% (Alshamiri et al., 2018; Basit et al., 2020).

Current medicants used for lipid regulation can have significant side effects, necessitating an exploration of alternative strategies. Natural treatments offer a promising avenue for reducing hyperlipidemia risks (Chattopadhyaya et al., 1996; Krause & Newton, 1995). Phenolic acids, flavonoids, epiafzelechin, epicatechin, saponins, procyanidin, proanthocyanidins, and anthocyanins have strong potential to regulate blood lipids (Nagpal et al., 2011). The dietary intake of 5%–10% polyphenols, flavonoids, and anthocyanins of diet for 21 days significantly regulated dyslipidemia in rats fed on high‐cholesterol diet (Queiroz et al., 2017). Oral intake of a polyphenol‐rich extract of 0.47 and 1.87 g/kg bw for 4 weeks reduced serum TG in HFD‐induced diabetic rats (Li et al., 2015). Flavonoids at a dose of 10 mg/kg bw per day also inhibited cholesterogenesis in atherogenic animals (Park et al., 2015). Procyanidins intake of 0.1 g/kg bw on a daily basis for 24 weeks offered positive effects on atherosclerosis and hyperlipidemia in HFD‐fed mice through lipid metabolic and liver cholesterol synthesis (Rong et al., 2017).

Thus, plants rich in phytochemicals have been used since ancient times for treatment of various diseases. C. fistula is one of the trees that has been traditionally used for therapeutic purposes. Its barks, leaves, and fruits (pods) have been used for many diseases. C. fistula pod contains a high amount of epiafzelechin, epicatechin, procyanidin, and saponins and can have protective role in metabolic disorders (Kashiwada et al., 1990; Mwangi et al., 2021). Therefore, the current research aimed to evaluate the phytochemical profile and therapeutic potential of C. fistula pod extract in hyperlipidemia.

2. METHODS AND MATERIALS

2.1. Procurement of raw material

Cassia fistula pods were procured from the local market, cleaned, and dried under sunlight. The pods were ground in a laboratory mill and stored for further analysis.

2.2. Extract preparation

A 500 g powder of C. fistula pods was immersed in 200 mL distilled water and was left for the night. Then, it was boiled for 3–4 h and further extraction occurred by using ultrasonic‐assisted extraction (UAE) method accordingly (Yingngam et al., 2019). For this purpose, the mixture was placed in a 400 mL beaker, placed in an ultrasonic water bath (Branson 3210, USA), and subjected to UAE treatment for 20 min at 45°C and 20% of amplitude. The decoction was cooled for 30 min at room temperature and then filtered through muslin cloth and further concentrated using a rotary evaporator at 50°C (Galviz‐Quezada et al., 2019).

2.3. Proximate analysis

The proximate analysis was performed using Association of Official Analytical Chemists (AOAC, 2005). Total nitrogen content was determined by the Kjeldahl method (Method# 976.05) and protein was calculated by using a factor of 6.25. Soxhlet method (Method# 920.39) was used for crude fat. Crude fiber was obtained after sample digestion (Method 962.09). Moisture was calculated after oven drying at 60°C for 24 h (Method 925.10). Ash was determined after heating at 550°C till ashing (Method 942.05).

2.4. Determination of phytochemicals

2.4.1. Total phenolic contents (TPC)

Folin–Ciocalteu was used for TPC accordingly (Singleton et al., 1999). The sample (0.5 mL) was poured into a tube containing 0.1 mL of Folin–Ciocalteu reagent. Then, 1.0 mL (7%) Na2CO3 was added and incubated for 1 h. The absorbance was recorded at 750 nm. A standard solution was prepared by dissolving 3 mg of gallic acid and a series of dilutions was made by using distilled water. Results are presented as mg gallic acid/g dry weight sample.

2.4.2. Total flavonoid content

Total flavonoid content (TFC) were determined using the method of Aiyegoro and Okoh (2010) with small modifications. Standard solution was prepared by dissolving 10 mg quercetin in methanol and dilutions (0, 2, 4, 6, 8, 10 μg/mL) were prepared to obtain the standard curve. A 1.0 mL sample, 4 mL of distilled water, and 0.3 mL of a 5% NaNO2 solution were mixed and incubated for 10 min. Following a 10‐min interval, 0.3 mL of a 10% AlCl3 solution was added. Subsequently, 2 mL of a 1.0 M NaOH solution was incorporated and making a total volume of 10 mL and absorbance was measured by UV spectrophotometer. TFCs were calculated from standard curve extrapolation and expressed as mg quercetin/g dry weight sample.

2.4.3. Condense tannin content (CT)

For CT contents determination, 0.5 mL of sample, 3 mL of vanillin solution (4%), and 1.5 mL of HCl were mixed for 10 min. The absorbance was recorded at 500 nm (Lister & Wilson, 2001). CT contents were measured with gallic acid standard and the results were expressed as mg gallic acid/g dry weight sample.

2.4.4. Saponins extraction/determination

The extraction/determination of saponins was determined through the procedure of Mora‐Ocación et al. (2022). First, 20 g of the sample was defatted with 300 mL petroleum ether and then was air‐dried. A volume of 300 mL methanol was used for the extraction of saponins from a sample that was kept overnight, and then 200 mL of water‐butanol (1:1 v/v) was mixed with methanol extract. It was further extracted with 50 mL of acetone to obtain saponins. A volume of 1.0 mL of standard solution and 3.5 mL of Liebermann‐Burchard reagent was used for optimal wavelength determination. Standard dilutions (0–0.4 mg/mL) and 1.0 mL of reagent were mixed and incubated for 30 min and absorbance was measured.

2.5. Antioxidant activity

DPPH and FRAP assay were used to determine the antioxidant activity of C. fistula extract.

2.5.1. DPPH assay

To prepare DPPH solution, 7.89 mg of DPPH was dissolved in 100 mL of absolute ethanol. The solution was then stored in darkness for 2 h. The 1.0 mL DPPH solution and 0.8 mL Tris–HCl were mixed and then 0.2 mL sample solution was added and incubated for 30 min at room temperature. The absorbance at 517 nm was recorded. Blank reading was taken using only 1.2 mL of ethanol and 0.8 mL of Tris–HCl Results were expressed as EC50 DPPH activity. EC50 was determined by plotting inhibition ratios (y) against sample concentrations (x) at six data points and fitting a regression line (y = ax + b). EC50 value was calculated by interpolating between two points where inhibition reached approximately 50%, and drawing a straight line to achieve 50% inhibition at the intersection point, forming another regression line (Y = AX + B). The concentration of the sample (X) corresponds to 50% inhibition using equation Y = AX + B (Valko et al., 2007). The inhibition ratio was calculated as:

DPPH%inhibition=control absorbance−sample absorbance/control absorbance×100

2.5.2. FRAP assay

The FRAP activity was measured using Benzie and Strain (1996) method. To prepare the FRAP solution, 10 mM TPTZ and 20 mM ferric chloride were combined in a 0.25 M acetate buffer with a pH of 3.6. Then, 0.15 mL of sample was mixed with 400 μL of water, and after 5 min, 3 mL of FRAP reagent was mixed. The absorbance was taken at 593 nm and the results were calculated as nmol Fe2+ equivalent/g dry extract.

3. EXPERIMENTAL DESIGN

Twenty‐four healthy female rats were weighed 190–200 g and kept in an animal room of the facility of the Department of Physiology, Government College University Faisalabad. Rats were caged individually with proper clean bedding and controlled with specific temperature and light. The whole research was conducted according to the ethical guidelines of the laboratory animal care (Gonder & Laber, 2007).

3.1. High‐fat diet (HFD) and induction of hyperlipidemia

Induction of hyperlipidemia in experimental animals was done using HFD {fat (58%), carbohydrate (17%), protein (25%) of total kcal} according to the procedure of Jain and Surana (2016) for 30 days. Induction was verified through lipid profile according to the criteria of “National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) (2001).” While control group rats were given a standard diet along with ad libitum throughout the trial.

3.2. Treatment plan

Rats were divided into four groups and each group contained six rats and C. fistula extract administration plan is shown in Table 1. C. fistula extract doses of 0.5 and 1.0 g /kg body weight were administrated through gavage. The doses were decided from a previous study (Abid et al., 2016). The intervention was carried out for a period of 1 month.

TABLE 1.

Treatment plan.

Group 1 Not induced rats Standard diet
Group 2 Induced rats Standard diet
Group 3 Induced rats Standard diet + 0.5 g extract
Group 4 Induced rats Standard diet + 1.0 g extract
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3.3. Data collection

The data was collected for the following parameters during the study.

3.3.1. Feed and water intake

The food intake of experimental rats was calculated on a daily basis by subtracting leftovers from the total diet offered and water was given through graduated drinking bottles and its intake was measured on a daily basis (Ahmad et al., 2018).

3.3.2. Body weight

The body weight of rats was measured on a weekly basis to determine the impact of treatment on body weight by using a digital weighing scale (QERINKLE).

3.3.3. Collection of blood sample

After the end of the trial, overnight fasting rats were decapitated, and blood samples were taken in a yellow‐coated tube. Centrifuged at RCF (g): 1252 for 10 min (SCILOGEX DM0636, Model # 91502303999) and serum was separated and frozen for further analysis.

3.3.4. Serum lipid analysis

Total cholesterol (TC) and TG were analyzed by enzymatic method following the modified protocol of Kim et al. (2011). HDL and LDL were assessed by the enzymatic method followed by the modifying protocol of Alshatwi et al. (2010).

3.3.5. Hepatic safety test

Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured by Sigma Aldrich Kit. Both tests were performed according to the instructions mentioned by the manufacturer.

3.3.6. Hematological parameters

Blood samples were collected and used to analyze the hematological parameters such as Red blood cell (RBC), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets, monocytes, and neutrophil, hemoglobin (Hb), using Biochem analyzer (Mindray Auto Hematology Analyzer, BC‐5200, USA).

3.3.7. Histopathological study

Liver samples were fixed in 10% formalin, followed by embedding in paraffin and sectioning to a thickness of 5 mm for histological analysis. These sections were then stained with hematoxylin and eosin for further examination. Pathological images were recorded at 10× magnification using a light microscope (Model BA410E Carlsbad, USA). It was evaluated using the method of Bernet et al. (1999).

3.4. Statistical analysis

Triplicate data in an Excel sheet was used for data calculation. Mean ± standard deviation results are presented. One‐way ANOVA (Statistix version 10) and the least significant difference (LSD) test were used to examine group differences. The p < .05 was used as significant.

4. RESULTS AND DISCUSSION

4.1. Extraction and physiochemical and antioxidant characterization

The UAE extraction, proximate analysis, phytochemicals analysis, and antioxidant activity results are presented in Table 2. The yield of extract from C. fistula pods was 25 ± 0.04%. The extract on a dry basis had 2.48 ± 0.15% moisture, 1.25 ± 0.55% fat, 11.93 ± 1.55% protein, 4.59 ± 0.55% fiber, 2.80 ± 0.20% ash, and 76.95 ± 0.55% NFE, respectively. The amount of TPC, TFC, CT, and saponins in the extract were 13.07 + 0.14 mg/g, 5.24 + 0.02 mg QE/g, 4.01 ± 0.52 mg/g, and 27.55 ± 0.59%, respectively. The antioxidant activity of DPPH and FRAP of the extract were 63.30 ± 0.015 μg/mL and 15.02 ± 0.34 nmol/g, respectively.

TABLE 2.

Compositional and physiochemical analysis of C. fistula extract.

Physiochemical analysis Parameters Plant extract
UAE extraction Yield 25 ± 0.04%
Proximate analysis Moisture (%) 2.48 ± 0.15
Fiber (%) 4.59 ± 0.55
Protein (%) 11.93 ± 1.55
Ash (%) 2.80 + 0.20
Fat (%) 1.25 ± 0.55
NEF (%) 76.95 ± 0.50
Phytochemicals analysis TPC (mg GAE/g) 13.07 ± 0.14
TFC (mg QE/g) 5.24 ± 0.02
CT (mg/g) 4.01 ± 0.52
Saponins (%) 27.55 ± 0.59
Antioxidant activity DPPH EC50 μg/mL 63.30 ± 0.015
FRAP (nmol/g) 15.02 ± 0.34
Ascorbic acid μg/mL 0.87 ± 0.02
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4.2. Biological evaluation of extract supplementation in vivo

The results of feed and water intake are presented in Tables 3 and 4. Maximum feed intake was observed in group 2 (30.99 ± 2.13 g) at the 4th week while the minimum feed intake was seen in group 4 (19.88 + 1.91 g) at the 3rd week, respectively.

TABLE 3.

Feed intake (g).

Time period Control Experiential
Weeks Group 1 Group 2 Group 3 Group 4
1st week 21.04 ± 0.23de 23.55 ± 3.17cd 26.22 ± 2.17bc 24.55 ± 1.13c
2nd week 21.27 ± 0.25de 27.83 ± 1.86b 26.94 ± 1.10b 21.00 ± 0.5de
3rd week 22.05 ± 0.5de 30.77 ± 1.60a 25.44 ± 2.08bc 19.88± 1.91e
4th week 22.55 ± 0.94de 30.99 ± 2.13a 21.83 ± 0.60de 22.16 ± 2.07de
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Note: Different superscript alphabets in the table are significantly different (p ≥ .05).

TABLE 4.

Water intake (mL).

Weeks Group 1 Group 2 Group 3 Group 4
1st week 18.92 ± 1.31e 22.00 ± 0.5cd 29.22 ± 0.34a 29.55 ± 0.25a
2nd week 22.05 ± 2.56cd 23.77 ± 1.07c 25.44 ± 0. 91b 24.16 ± 0.83bc
3rd week 22.66 ± 0.60cd 21.38 ± 0.19d 22.38 ± 0.55cd 21.38 ± 1.63d
4th week 23.11 ± 3.15c 25.00 ± 1.01b 21.22 ± 0.25d 20.05 ± 0.42d
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Note: Different superscript alphabets in the table are significantly different (p ≥ .05).

The highest water intake (29.55 ± 0.25 mL) was observed in group 4 in the 1st week and the lowest water intake (20.05 ± 0.42 mL) was observed in the 4th week in the same group.

The body weight changes of the rat groups are shown in Table 5. According to the results, the highest body weight change was observed in group 4 while the lowest change was observed in group 2.

TABLE 5.

Weight change (g).

Weeks Group 1 Group 2 Group 3 Group 4
1st week 239.83 ± 10.33c 248.83 ± 9.49b 261.33 ± 10.05 a 251.00 ± 13.92b
2nd week 215.00 ± 2.88de 224.33 ± 8.89d 233.00 ± 12.87cd 213.33 ± 9.22e
3rd week 206.17 ± 4.16ef 217.00b ± 6.44de 189.33 ± 13.10f 177.00 ± 12.40
4th week 182.50 ± 10.44f 204.50 ± 8.04ef 155.50 ± 6.56g 147.83 ± 6.9g
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Note: Different superscript alphabets in the table are significantly different (p ≥ .05).

The values of lipids are presented in Table 6. Results have shown a significant (p = .00) decrease in TC, TG, and LDL of groups 3 and 4 as compared to group 2. HDL has increased significantly (p = .00) in groups 3 and 4 as compared to group 2.

TABLE 6.

Effect of C. fistula extract on the lipid profile of hyperlipidemic rats.

Experimental groups Lipid parameters (mg/dL) F value
Group 1 Group 2 Group 3 Group 4 p Value
TC 73.33 ± 7.20 150.01 ± 6.16 115.33 ± 5.06 90.83 ± 8.86 .00** 131.26
TG 84.50 ± 7.36 119.67 ± 10.98 101.67 ± 9.33 74.16 ± 9.10 .00** 27.76
HDL 20.83 ± 2.99 17.33 ± 4.50 40.76 ± 4.38 41.83 ± 8.4 .00** 33.62
LDL 72.50 ± 6.53 114.00 ± 7.29 76.33 ± 4.32 74.83 ± 4.66 .00** 68.91
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Table 7 shows the result of hepatic enzymes after intake of C. fistula extract. AST and ALT values were significantly (p = .00) reduced in groups 3 and 4 as compared to group 2.

TABLE 7.

Effect of C. fistula extract on liver enzymes of hyperlipidemic rats.

Parameters Control Experimental groups
Group 1 Group 2 Group 3 Group 4 p Value F value
AST (U/L) 57.083 ± 10.89 97.25 ± 12.93 73.00 ± 5.47 67.16 ± 10.60 .00** 16.31
ALT (U/L) 25.66 ± 3.50 56.50 ± 11.77 25.58 ± 4.56 22.33 ± 7.22 .00** 27.63
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The hematological results are presented in Table 8. The HB, RBC, MCHC, and MCV were significantly (p = .00) higher in groups 3 and 4 as compared to group 2. Neutrophils, monocytes, and platelets were reduced significantly (p = .00) in both groups as compared to group 2.

TABLE 8.

Effect on hematological parameters.

Parameters Control Experimental groups Significance
Group 1 Group 2 Group 3 Group 4 p Value F value
HB (g/dL) 11.045 ± 0.43 6.75 ± 0.80 11.06 ± 0.69 11.37 ± 0.43 .00** 81.37
RBCs (106/μL) 8.145 ± 1.00 6.47 ± 0.36 7.81 ± 0.64 8.03 ± 0.67 .00** 6.94
MCHC (g/dL) 36.33 ± 3.93 23.67 ± 3.31 34.69 ± 0.62 35.02 ± 1.78 .00** 27.64
MCH (Pg) 17.31 ± 0.29 15.96 ± 2.41 17.29 ± 0.71 18.00 ± 0.26 .06 2.91
MCV (fL) 52.44 ± 3.49 42.52 ± 7.06 54.01 ± 1.77 55.36 ± 4.01 .00** 10.01
Neutrophil (%) 11.76 ± 2.32 36.35 ± 1.86 17.19 ± 8.55 11.23 ± 0.51 .00** 40.43
Monocytes (%) 2.033 ± 0.85 4.38 ± 0.21 2.13 ± 1.12 2.01 ± 0.63 .00** 12.81
Platelet (103/μL) 848.67 ± 96.61 990.33 ± 90.03 765.67 ± 57.78 755.00 ± 32.38 .00** 12.60
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Figure 1 illustrates the histopathological changes in the liver. The extension of a pathological change as rated ‘score value’ and the pathological importance of this alteration as ‘importance factor’ were collectively used to categorize the changes. According to the score, architectural and structural alteration with fat deposits and necrosis was observed. Histopathology images of fatty liver showed fat accumulation as black arrows and necrosis as blue arrows induced by HFD‐induced hyperlipidic rats in Figure 1b. However, fat accumulation and inflammatory necrosis reduction in dose‐dependent manner were observed in HFD‐induced hyperlipidic rats shown in Figure 1c,d which indicates the protective effect of C. fistula extract. Figure 1a is non‐HFD‐induced untreated control group showing normal structure of liver histology.

FIGURE 1.

FIGURE 1

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Histopathology images of liver at 40× magnification fixed with 10% formalin and stained with hematoxylin for fatty liver analysis. The fat accumulation in black arrows and necrosis in blue arrows induced by HFD‐induced hyperlipidic rats are shown in (b) while the protective effect of C. fistula extract in HFD‐induced hyperlipidic rats is shown in (c and d). (a) is non‐HFD‐induced untreated control group.

5. DISCUSSION

The proximal composition analysis of pods aligned with prior studies (Kdam, 2022; Shukla et al., 2013), yet TPC and TFC were notably higher than reported values by previous studies (Chourasia et al., 2019) and potentially attributed to variations in plant part's utilization. Comparisons with the results of Irshad et al. (2012) revealed discrepancies in TPC and TFC levels, possibly due to differences in extracting solvents and plant part's utilization. Such variations in phytochemical composition across studies may stem from various geographic regions and variations in plant nutrition. Our assessments encompassed yield extraction, proximal analysis, phytochemical content, and antioxidant activity showed that notably water and UAE extraction methods yield superior results compared to methanol/ethanol extraction and consistent with findings by Tan et al. (2018). Assessment of antioxidant activity revealed promising results, consistent with prior findings by Abdellatif et al. (2024). These results suggest that C. fistula extract possesses potent antioxidant properties, attributed to specific phytochemical constituents. Observations on experimental groups indicated significant variations in feed intake over time, potentially influenced by hyperlipidemia effects. Notably, reductions in feed intake correlated with C. fistula extract consumption, consistent with reports by Shah et al. (2019). Distinct patterns of water intake, with higher initial levels followed by gradual reductions, may reflect metabolic differences and dietary fiber content, as observed in hyperlipidemic rats (Tamargo et al., 2020). Significant reductions in body weight among hyperlipidemic groups suggest metabolic alterations associated with C. fistula extract consumption, supporting previous studies indicating its role in suppressing cholesterol and TG biosynthesis (Abid et al., 2016; Jangir & Jain, 2017).

The efficiency of C. fistula extract in modulating lipid metabolism and exerting antioxidant effects elucidated its potential mechanisms underlying the observed effects, attributing reductions in LDL levels to enhanced hepatic LDL receptor binding facilitated by polyphenolic extracts (Kaur et al., 2019). Moreover, increases in HDL levels may result from enhanced lecithin‐cholesterol acyltransferase activity, promoting reverse cholesterol transport and endothelial protection (Ossoli et al., 2016). C. fistula's hepatoprotogenic effects were evidenced by decreased AST and ALT levels, attributed to its antioxidant compounds, consistent with reports by Kaur et al. (2019). These findings underscore the potential of C. fistula extract as a hepatoprotogerative agent. The study highlighted immunomodulatory effects, with flavonoids such as quercetin enhancing erythropoietin activity, leading to elevated neutrophil, monocyte, and platelet levels in hyperlipidemic rats. Notably, C. fistula extract mitigated these effects, suggesting its potential in ameliorating oxidative stress‐induced hematological alterations (Abdellatif et al., 2024; Straface et al., 2010). While reductions in neutrophil and monocyte levels attributed to flavonoid extracts, align with prior studies indicating their role in mitigating stress‐induced immune responses (Lin et al., 2022). Moreover, decreases in platelet counts suggest potential antiplatelet effects, indicative of vascular disease prevention (Asgary et al., 2012). Histopathological examination provides valuable insights into the structural alteration induced by hyperlipidemia and the protective effects of C. fistula extract. Histological analysis of liver tissue revealed lipid accumulation, hepatocellular injury, and inflammatory infiltrates as necrosis characteristic of hyperlipidemic conditions. The current study's histopathological findings may exhibit improvements in liver architecture following C. fistula extract administration, characterized by reduced lipid accumulation, hepatocellular, and inflammatory changes as necrosis. These observations are consistent with previous reports indicating the hepatoprotective effects of C. fistula against induced liver damage (Thompson et al., 2017). The antioxidant and hepatoprotective effects underscore the consistent pharmacological potential of C. fistula and can be used as lipid‐lowering agents in hyperlipidemia.

6. CONCLUSION

The outcome of the current study demonstrated that C. fistula extract is an excellent source of polyphenol compound and has strong antioxidant potential that can provide probable benefits to consumers. Results of the efficacy study have shown anti‐hyperlipidemic effects of extract of C. fistula. It decreases TC, TG, LDL, and HDL in hyperlipidemic rats.

AUTHOR CONTRIBUTIONS

Maryam Tariq: Conceptualization (equal); data curation (equal). Nazir Ahmad: Project administration (equal); supervision (equal); writing – original draft (equal). Mahr Un Nisa: Validation (equal). Muhammad Abdul Rahim: Funding acquisition (equal); writing – review and editing (equal). Eliasse Zongo: Funding acquisition (equal); writing – review and editing (equal).

FUNDING INFORMATION

No specific grant was used in this study.

CONFLICT OF INTEREST STATEMENT

No competing interests for any purpose are declared by the authors.

ETHICAL APPROVAL

Government College University Faisalabad intuitional ethical approval was obtained from the ethic review committee under registration number Ref. No. GCUF.ERC/23.

ACKNOWLEDGMENTS

We are thankful to the digital library of GCUF for providing access to research data. The authors are also grateful to Times Institute, Multan, Pakistan, for helping in providing resources for literature.

Tariq, M. , Ahmad, N. , Nisa, M. U. , Rahim, M. A. , & Zongo, E. (2024). Phytochemicals profiling of Cassia fistula fruit extract and its effect on serum lipids and hematological parameters in high‐fat diet‐induced hyperlipidemic female rats. Food Science & Nutrition, 12, 5776–5784. 10.1002/fsn3.4229

Contributor Information

Nazir Ahmad, Email: drnazirahmad@gcuf.edu.pk.

Muhammad Abdul Rahim, Email: abdul.rahim@gcuf.edu.pk, Email: mabdulrahim@t.edu.pk.

Eliasse Zongo, Email: zongoeliasse@yahoo.fr.

DATA AVAILABILITY STATEMENT

It is original and unpublished research data and will be available as per publisher policy.

REFERENCES

  1. Abdellatif, N. A. , Eltamany, E. E. , El‐Shenawy, N. S. , Nafie, M. S. , Hassan, Y. M. , Al‐Eisa, R. A. , & Abdelhameed, R. F. (2024). Cassia fistula leaves extract profiling and its emphasis on induced ulcerative colitis in male rats through inhibition of caspase 3 and cyclooxygenase‐2. Arabian Journal of Chemistry, 17, 105672. [Google Scholar]
  2. Abid, R. , Mahmood, R. , & Santosh Kumar, H. S. (2016). Hypolipidemic and antioxidant effects of ethanol extract of Cassia fistula fruit in hyperlipidemic mice. Pharmaceutical Biology, 54(12), 2822–2829. [DOI] [PubMed] [Google Scholar]
  3. Ahmad, U. , Ahmad, R. S. , Arshad, M. S. , Mushtaq, Z. , Hussain, S. M. , & Hameed, A. (2018). Antihyperlipidemic efficacy of aqueous extract of Stevia rebaudiana Bertoni in albino rats. Lipids in Health and Disease, 17, 1–8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Aiyegoro, O. A. , & Okoh, A. I. (2010). Preliminary phytochemical screening and in vitro antioxidant activities of the aqueous extract of Helichrysum longifolium DC. BMC Complementary and Alternative Medicine, 10, 1–8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Alshamiri, M. , Ghanaim, M. M. A. , Barter, P. , Chang, K. C. , Li, J. J. , Matawaran, B. J. , & Yusof, A. K. (2018). Expert opinion on the applicability of dyslipidemia guidelines in Asia and the Middle East. International Journal of General Medicine, 11, 313–322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Alshatwi, A. A. , Al Obaaid, M. A. , Al Sedairy, S. A. , Al‐Assaf, A. H. , Zhang, J. J. , & Lei, K. Y. (2010). Tomato powder is more protective than lycopene supplement against lipid peroxidation in rats. Nutrition Research, 30(1), 66–73. [DOI] [PubMed] [Google Scholar]
  7. Aoac, C. (2005). Official methods of analysis of the Association of Analytical Chemists International. Official Methods. [Google Scholar]
  8. Asgary, S. , Najafi, S. , Ghannadi, A. , Dashti, G. , & Helalat, A. (2012). Efficiency of black cumin seeds on hematological factors in normal and hypercholesterolemic rabbits. ARYA Atherosclerosis, 7(4), 146. [PMC free article] [PubMed] [Google Scholar]
  9. Basit, A. , Sabir, S. , Riaz, M. , & Fawwad, A. (2020). NDSP 05: Prevalence and pattern of dyslipidemia in urban and rural areas of Pakistan; a sub analysis from second National Diabetes Survey of Pakistan (NDSP) 2016–2017. Journal of Diabetes & Metabolic Disorders, 19, 1215–1225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Benzie, I. F. , & Strain, J. J. (1996). The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: The FRAP assay. Analytical Biochemistry, 239(1), 70–76. [DOI] [PubMed] [Google Scholar]
  11. Bernet, D. , Schmidt, H. , Meier, W. , Burkhardt‐Holm, P. , & Wahli, T. (1999). Histopathology in fish: Proposal for a protocol to assess aquatic pollution. Journal of Fish Diseases, 22(1), 25–34. [Google Scholar]
  12. Cao, X. , Zhang, L. , Chen, C. , Wang, Q. , Guo, L. , Ma, Q. , & Li, J. (2017). The critical role of ABCG1 and PPARγ/LXRα signaling in TLR4 mediates inflammatory responses and lipid accumulation in vascular smooth muscle cells. Cell and Tissue Research, 368, 145–157. [DOI] [PubMed] [Google Scholar]
  13. Chattopadhyaya, R. , Pathak, D. , & Jindal, D. P. (1996). Antihyperlipidemic agents: A review. Indian Drugs, 33(3), 85–97. [Google Scholar]
  14. Chourasia, V. , Ahmad, S. , Parmar, A. , Kulkarni, S. , & Sahu, N. K. (2019). Evaluation of anti‐hyperlipidemic activity of ethanolic extract of Cassia fistula leaf. Journal of Drug Delivery and Therapeutics, 9(4‐A), 685–693. [Google Scholar]
  15. Expert Panel on Detection, E . (2001). Executive summary of the third report of the National Cholesterol Education Program (NCEP) expert panel on detection, evaluation, and treatment of high blood cholesterol in adults (adult treatment panel III). JAMA, 285(19), 2486–2497. [DOI] [PubMed] [Google Scholar]
  16. Fioranelli, M. , Bottaccioli, A. G. , Bottaccioli, F. , Bianchi, M. , Rovesti, M. , & Roccia, M. G. (2018). Stress and inflammation in coronary artery disease: a review psychoneuroendocrineimmunology‐based. Frontiers in immunology, 9, 358054. 10.3389/fimmu.2018.02031 [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Galviz‐Quezada, A. , Ochoa‐Aristizábal, A. M. , Zabala, M. E. A. , Ochoa, S. , & Osorio‐Tobón, J. F. (2019). Valorization of iraca (Carludovica palmata, Ruiz & pav.) infructescence by ultrasound‐assisted extraction: An economic evaluation. Food and Bioproducts Processing, 118, 91–102. [Google Scholar]
  18. Gonder, J. C. , & Laber, K. (2007). A renewed look at laboratory rodent housing and management. ILAR Journal, 48(1), 29–36. [DOI] [PubMed] [Google Scholar]
  19. Howell, K. W. , Meng, X. , Fullerton, D. A. , Jin, C. , Reece, T. B. , & Cleveland, J. C., Jr. (2011). Toll‐like receptor 4 mediates oxidized LDL‐induced macrophage differentiation to foam cells. Journal of Surgical Research, 171(1), e27–e31. [DOI] [PubMed] [Google Scholar]
  20. Irshad, M. , Zafaryab, M. D. , Singh, M. , & Rizvi, M. (2012). Comparative analysis of the antioxidant activity of Cassia fistula extracts. International Journal of Medicinal Chemistry, 2012, 157125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jaffe, I. Z. , & Karumanchi, S. A. (2024). Lipid droplets in the endothelium: The missing link between metabolic syndrome and cardiovascular disease? Journal of Clinical Investigation, 134(4), e176347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jain, P. G. , & Surana, S. J. (2016). Isolation, characterization and hypolipidemic activity of ferulic acid in high‐fat‐diet‐induced hyperlipidemia in laboratory rats. EXCLI Journal, 15, 599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jangir, R. N. , & Jain, G. C. (2017). Evaluation of antidiabetic activity of hydroalcoholic extract of Cassia fistula Linn. pod in streptozotocin‐induced diabetic rats. Pharmacognosy Journal, 9(5), 599–606. [Google Scholar]
  24. Kashiwada, Y. , Iizuka, H. , Yoshioka, K. , Chen, R. F. , Nonaka, G. I. , & Nishioka, I. (1990). Tannins and related compounds. XCIII.: Occurrence of enantiomeric proanthocyanidins in the leguminosae plants, Cassia fistula L. and C. javanica L. Chemical and Pharmaceutical Bulletin, 38(4), 888–893. [Google Scholar]
  25. Kaur, S. , Sharma, D. , Singh, A. P. , & Kaur, S. (2019). Amelioration of hepatic function, oxidative stress, and histopathologic damages by Cassia fistula L. fraction in thioacetamide‐induced liver toxicity. Environmental Science and Pollution Research, 26, 29930–29945. [DOI] [PubMed] [Google Scholar]
  26. Kdam, R. M. G. (2022). Proximate analysis and phytochemical screening of Golden shower tree (Cassia fistula L.) leaves and fruits and its antimicrobial activity (Doctoral dissertation, University of Gezira) .
  27. Kim, J. Y. , Paik, J. K. , Kim, O. Y. , Park, H. W. , Lee, J. H. , Jang, Y. , & Lee, J. H. (2011). Effects of lycopene supplementation on oxidative stress and markers of endothelial function in healthy men. Atherosclerosis, 215(1), 189–195. [DOI] [PubMed] [Google Scholar]
  28. Krause, B. R. , & Newton, R. S. (1995). Lipid‐lowering activity of atorvastatin and lovastatin in rodent species: Triglyceride‐lowering in rats correlates with efficacy in LDL animal models. Atherosclerosis, 117(2), 237–244. [DOI] [PubMed] [Google Scholar]
  29. Li, C. Q. , Liao, X. B. , Li, X. H. , Guo, J. W. , Qu, X. L. , & Li, L. M. (2015). Effect and mechanism of Litchi semen effective constituents on insulin resistance in rats with type 2 diabetes mellitus. Zhong Yao Cai= Zhongyaocai= Journal of Chinese Medicinal Materials, 38(7), 1466–1471. [PubMed] [Google Scholar]
  30. Lin, Z. , Gan, T. , Huang, Y. , Bao, L. , Liu, S. , Cui, X. , & Qian, Y. (2022). Anti‐inflammatory activity of mulberry leaf flavonoids in vitro and in vivo. International Journal of Molecular Sciences, 23(14), 7694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lister, E. , & Wilson, P. (2001). Measurement of total phenolics and ABTS assay for antioxidant activity (personal communication). Crop Research Institute, Lincoln, New Zealand, 7, 235–239. [Google Scholar]
  32. Mora‐Ocación, M. S. , Morillo‐Coronado, A. C. , & Manjarres‐Hernández, E. H. (2022). Extraction and quantification of saponins in quinoa (Chenopodium quinoa Willd.) genotypes from Colombia. International Journal of Food Science, 2022, 7287487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mwangi, R. W. , Macharia, J. M. , Wagara, I. N. , & Bence, R. L. (2021). The medicinal properties of Cassia fistula L: A review. Biomedicine & Pharmacotherapy, 144, 112240. [DOI] [PubMed] [Google Scholar]
  34. Nagpal, M. A. , Nagpal, N. , Rahar, S. , Shah, G. , Swami, G. , & Kapoor, R. (2011). Phytochemical investigation of methanolic extract of Cassia fistula leaves. Pharmacognosy Journal, 3(26), 61–69. [Google Scholar]
  35. Naser, I. H. , Alkareem, Z. A. , & Mosa, A. U. (2021). Hyperlipidemia: Pathophysiology, causes, complications, and treatment. A review. Karbala Journal of Pharmaceutical Sciences, 1(19), 118–132. [Google Scholar]
  36. Ossoli, A. , Pavanello, C. , & Calabresi, L. (2016). High‐density lipoprotein, lecithin: Cholesterol acyltransferase, and atherosclerosis. Endocrinology and Metabolism, 31(2), 223–229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Park, C. H. , Noh, J. S. , Fujii, H. , Roh, S. S. , Song, Y. O. , Choi, J. S. , & Yokozawa, T. (2015). Oligonol, a low‐molecular‐weight polyphenol derived from lychee fruit, attenuates gluco‐lipotoxicity‐mediated renal disorder in type 2 diabetic db/db mice. Drug Discoveries & Therapeutics, 9(1), 13–22. [DOI] [PubMed] [Google Scholar]
  38. Queiroz, E. R. , Abreu, C. M. D. , Rocha, D. A. , Sousa, R. V. , Fraguas, R. M. , Braga, M. A. , & Cesar, P. H. (2017). Lychee (Litchi chinensis Sonn.) peel flour: Effects on hepatoprotection and dyslipidemia induced by a hypercholesterolemic diet. Anais da Academia Brasileira de Ciências, 90, 267–281. [DOI] [PubMed] [Google Scholar]
  39. Rafaqat, S. , Rafaqat, S. , & Klisić, A. (2024). The role of serum lipid profile in the pathogenesis of arterial hypertension. Archives of Pharmacy, 74(Notebook 1), 76–91. [Google Scholar]
  40. Rong, S. , Hu, X. , Zhao, S. , Zhao, Y. , Xiao, X. , Bao, W. , & Liu, L. (2017). Procyanidins extracted from the litchi pericarp ameliorate atherosclerosis in ApoE knockout mice: Their effects on nitric oxide bioavailability and oxidative stress. Food & Function, 8(11), 4210–4216. [DOI] [PubMed] [Google Scholar]
  41. Shah, M. N. , Iqbal, A. , Rıaz, M. , Khan, S. , Khan, M. T. , Sıal, A. R. , & Bayram, İ. (2019). Effect of graded supplementation levels of golden chain tree (Cassia fistula L.) leaves on feed intake, nutrients digestibility and plasma lipids profile in growing rabbits. Atatürk Üniversitesi Veteriner Bilimleri Dergisi, 14(3), 263–272. [Google Scholar]
  42. Shukla, R. K. , Porval, A. , Painuly, D. , & Shukla, A. (2013). Physico‐chemical characterstics, proximate analysis and total phenolic content of Cassia fistula bark. Nat. Prod. Indian J, 9, 133–137. [Google Scholar]
  43. Singleton, V. L. , Orthofer, R. , & Lamuela‐Raventós, R. M. (1999). [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin‐ciocalteu reagent. In Methods in enzymology (Vol. 299, pp. 152–178). Academic press. [Google Scholar]
  44. Straface, E. , Gambardella, L. , Metere, A. , Marchesi, A. , Palumbo, G. , Cortis, E. , & Del Principe, D. (2010). Oxidative stress and defective platelet apoptosis in naïve patients with Kawasaki disease. Biochemical and Biophysical Research Communications, 392(3), 426–430. [DOI] [PubMed] [Google Scholar]
  45. Tamargo, A. , Martin, D. , Del Hierro, J. N. , Moreno‐Arribas, M. V. , & Muñoz, L. A. (2020). Intake of soluble fibre from chia seed reduces bioaccessibility of lipids, cholesterol and glucose in the dynamic gastrointestinal model simgi®. Food Research International, 137, 109364. [DOI] [PubMed] [Google Scholar]
  46. Tan, B. L. , Norhaizan, M. E. , Liew, W. P. P. , & Sulaiman Rahman, H. (2018). Antioxidant and oxidative stress: a mutual interplay in age‐related diseases. Frontiers in pharmacology, 9, 402374. 10.3389/fphar.2018.01162 [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Thompson, M. , Jaiswal, Y. , Wang, I. , & Williams, L. (2017). Hepatotoxicity: Treatment, causes and applications of medicinal plants as therapeutic agents. The Journal of Phytopharmacology, 6(3), 186–193. [Google Scholar]
  48. Valko, M. , Leibfritz, D. , Moncol, J. , Cronin, M. T. , Mazur, M. , & Telser, J. (2007). Free radicals and antioxidants in normal physiological functions and human disease. The International Journal of Biochemistry & Cell Biology, 39(1), 44–84. [DOI] [PubMed] [Google Scholar]
  49. Yingngam, B. , Zhao, H. , Baolin, B. , Pongprom, N. , & Brantner, A. (2019). Optimization of ultrasonic‐assisted extraction and purification of Rhein from Cassia fistula pod pulp. Molecules, 24(10), 2013. [DOI] [PMC free article] [PubMed] [Google Scholar]

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