Open Veterinary Journal, (2025), Vol. 15(9): 4204-4209

Research Article

10.5455/OVJ.2025.v15.i9.26

Comparative effects of olive oil and Nigella sativa oil on mice lipid profiles

Lubna Ahmed Kafi^*

Department of Quality Assurance and University Performance, Ibn Sina University of Medical and Pharmaceutical Sciences, Baghdad, Iraq

*Corresponding Author: Lubna Ahmed Kafi. Ibn Sina University of Medical and Pharmaceutical Sciences, Baghdad, Iraq. Email: proflubnakafi [at] yahoo.com

Submitted: 14/05/2025 Revised: 24/07/2025 Accepted: 07/08/2025 Published: 30/09/2025

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ABSTRACT

Background: Olive oil and Nigella sativa oil are recognized for their lipid-lowering effects and antioxidant properties. Limited comparative data are available on their effects in mice without any dietary intervention.

Aim: This study assesses the impact of olive oil and N. sativa oil on the serum lipid profiles of mice without any other dietary intervention or hyperlipidemia induction.

Methods: Fifteen male Swiss mice were randomly divided into three groups of 5 each. The control group was orally administered with 5 ml/kg of body weight/day of sunflower oil for 60 consecutive days. The doses of olive oil and N. sativa oil were at 0.4 ml/kg of body weight/day for 60 consecutive days, orally, after mixing them separately with the vehicle, sunflower oil, up to 5 ml/kg. Twenty-four hours after the last treatment, blood samples were obtained from the heart of each mouse, and the serum was used for lipid profile analyses using analytical kits.

Results: Both olive oil and N. sativa oil significantly (p < 0.05) decreased serum levels of cholesterol, triglycerides, LDL, and VLDL in comparison with those of control values. Treatments significantly increased serum HDL levels compared with the control value. Changes in lipid profiles were not significantly different between the olive oil and N. sativa oil groups. The ratios of cholesterol, triglycerides, and LDL versus HDL of mice treated with olive oil and N. sativa oil were all significantly below the respective control values as a result of reducing lipid profiles and increasing HDL levels.

Conclusion: Olive oil and N. sativa oil can significantly improve blood lipid levels in mice, particularly by lowering atherogenic lipids and increasing HDL. These changes result in favorable cholesterol, triglyceride, and low-density lipoprotein/high-density lipoprotein ratios, which play an important role in lowering cardiovascular disease risk and highlight the potential therapeutic and possibly protective benefits of these oils in managing lipid disorders.

Keywords: Olive, Hyperlipidemia, Cholesterol, Triglycerides, High-density lipoprotein.

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Introduction

Traditional medicine has used plants and plant products for the prevention (Kafi and Kbyeh, 2020) and treatment of various pathological conditions, including hyperlipidemia (Kafi, 2014; Kafi and Al-Ezzi, 2015, 2017; Tarmoos and Kafi, 2019), diabetes (Kafi, 2016), inflammation (Kafi and AL-Zobahi, 2012, 2013), blood disorders (Al-Taee and Kafi, 2013), cancer (Jumaa et al., 2015; Kafi, 2015; Barakat et al., 2022), reproductive disorders (Al-Khayyat et al., 2016a,b), and wounds (Al-Khayyat et al., 2008).

Dyslipidemia is a risk factor for cardiovascular diseases and metabolic disorders. It is characterized by abnormal levels of blood lipids, particularly cholesterol, triglycerides, and lipoproteins (Alloubani et al., 2021). In addition to therapeutic medications available for hyperlipidemia (Wright and Subramanian, 2024; Wierzbicki, 2025), dietary interventions using natural oils, such as olive oil and Nigella sativa (black seed) powder or oil, have been used for this purpose (Ali and Blunden, 2003; Asgary et al., 2015; Finicelli et al., 2019; Farias-Pereira et al., 2023). They are recognized for their lipid-lowering effects and antioxidant properties (Ali and Blunden, 2003; Ahmad and Beg, 2013; Finicelli et al., 2019; Farias-Pereira et al., 2023).

Olive oil is a key component of the Mediterranean diet and is rich in monounsaturated fatty acids, such as oleic acid, and contains bioactive compounds that reduce inflammation and act as antioxidants (Finicelli et al., 2019; Farias-Pereira et al., 2023). Studies have shown that olive oil has a role in improving lipid profiles in both humans (Buckland and Gonzalez, 2015; Montserrat-de la Paz et al., 2016; Jabbarzadeh-Ganjeh et al., 2023) and experimental animals, such as mice and rats (Kafi, 2014; Katsarou et al., 2016; Albrahim et al., 2022). It has been reported that the beneficial effects are reflected by reducing total cholesterol, low-density lipoprotein cholesterol (LDL), very low-density lipoprotein cholesterol (VLDL), and triglycerides, as well as increasing the high-density lipoprotein cholesterol (HDL) in dyslipidemic patients and hyperlipidemic animal models (Kafi, 2014; Buckland and Gonzalez, 2015; Katsarou et al., 2016; Montserrat-de la Paz et al., 2016; Albrahim et al., 2022). For instance, in a study on mice fed a high-lipid diet, two months of oral olive oil administration significantly improved their serum lipid profiles, particularly increasing HDL levels compared to the control group (Kafi, 2014). Other studies in hyperlipidemic rats have shown that olive oil reduces total cholesterol, LDL, and triglycerides levels and significantly enhances antioxidant and anti-inflammatory functions (Katsarou et al., 2016; Albrahim et al., 2022).

N. sativa oil is made from black cumin seeds and has long been used in traditional medicine. It contains thymoquinone and other bioactive components (Ali and Blunden, 2003; Ahmad and Beg, 2013; Asgary et al., 2015). In studies with mice having high serum lipid levels, daily administration of N. sativa oil for up to 2 months significantly reduced total cholesterol, LDL, and triglyceride levels (Kafi, 2014). However, olive oil and N. sativa oil might act differently, especially in raising HDL (Ali and Blunden, 2003; Ahmad and Beg, 2013). A meta-analysis study in 2023 showed that olive oil had minor effects on total cholesterol, LDL, HDL, and triglycerides in people with normal lipid levels (Jabbarzadeh-Ganjeh et al., 2023). These differences may result from initial lipid levels, oil use duration, or species differences in expected responses between humans and animals. Despite this, the antioxidant and anti-inflammatory benefits of olive oil have been well documented, supporting liver integrity and reducing lipid peroxidation in animal studies (Kafi, 2014; Katsarou et al., 2016; Albrahim et al., 2022). The antioxidant properties of thymoquinone are mainly responsible for the effects of N. sativa (Ali and Blunden, 2003; Ahmad and Beg, 2013).

Considering the beneficial health effects of olive oil and N. sativa oil, additional explorations are needed on their roles in humans and experimental animals with normal lipid profiles. Because of the lack of direct comparison between these two oils in normolipidemic mice, this study aimed to evaluate the impact of daily administration of olive oil and N. sativa oil for 2 months on the serum lipid profiles of mice without any other dietary intervention or hyperlipidemia induction. The results of this research may provide important insights into the effects of these natural oils on the lipid profiles of mice under controlled experimental conditions. Additionally, these findings may have translational implications for humans.

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Materials and Methods

Fifteen male CD-1 Swiss adult mice (age 2–2.5 months; body weight 25–30 g) were used in this study. The mice were housed at a temperature between 20ºC and 25ºC and a 12-hour light/dark cycle, with water and laboratory food provided ad libitum.

Olive and N. sativa oils

Olive fruits and N. sativa seeds were used after identification by pharmacognosy experts at the Department of Pharmacology, College of Veterinary Medicine, University of Baghdad. The mature olive fruits were collected from local sources. The olive oil fraction was extracted after malaxation without heating according to a previously reported method (Kafi, 2014, 2015; Ferro et al., 2023). N. sativa oil seeds were purchased from local sources. The oil of N. sativa was also extracted as previously described (Pourbakhsh et al., 2014; Kafi, 2014; 2015).

Oil treatments

Mice were randomly divided into three groups of five animals each, and blinded treatment allocations were applied. The control group was orally administered with sunflower oil (Zer, Turkey) at 5 ml/kg of body weight/day for 60 consecutive days. The choice of sunflower oil was based on previous studies that showed that it is a good vehicle for other oily treatments (Kafi, 2014; Tarmoos and Kafi, 2019). The doses of olive oil and N. sativa oil were 0.4 ml/kg of body weight/day for 60 consecutive days, orally after mixing them separately with the vehicle, sunflower oil, to make the volume of administration at 5 ml/kg (Kafi, 2014). The mice were monitored daily for any side effects of treatment, if present. Twenty-four hours after the last oil treatment, blood samples were obtained from the heart of each mouse to obtain sufficient blood for the biochemical assays, and the serum was harvested for lipid profile analyses (Tarmoos and Kafi, 2019). The serum samples were subjected to lipid profile assays using analytical kits obtained from Linear Chemicals, Spain (Friedewald et al., 1972; Kafi, 2014).

Statistical analysis

Lipid profile data and the ratios of cholesterol, triglycerides, and LDL versus HDL in each group are presented as mean ± SE of five mice/treatment group. The statistical package Past5.1 (https://www.nhm.uio.no/english/research/resources/past/) was used to analyze the data by one-way analysis of variance, followed by Tukey’s pairwise comparison. The level of statistical significance was p < 0.05.

Ethical approval

The animal experiments were conducted according to institutional regulations and ethics in accordance with the guidelines of the National Research Council for Animal Care and Research (National Research Council, 2011) and Animal Research: Reporting of in vivo Experiments (ARRIVE) (Percie du Sert et al., 2020). The approvals of the Animal House and Department of Pharmacology at the College of Veterinary Medicine, University of Baghdad, Iraq were obtained to conduct this research in mice.

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Results

Treatment of mice with olive oil and N. sativa oil at 0.4 ml/kg/day for 60 days significantly (p < 0.05) decreased the serum levels of cholesterol, triglycerides, LDL, and VLDL compared with those of the control group (Table 1). Concurrently, treatment significantly increased the serum HDL levels compared with the control group. Changes in lipid profiles were not significantly different between the olive oil and N. sativa oil groups (Table 1). None of the mice showed adverse effects from the treatments, indicating the clinically beneficial effects of the treatment regimens.

Table 1. Lipid profiles of male mice orally treated with 0.4 ml/kg/day olive oil and Nigella sativa oil for 60 consecutive days.

The ratios of cholesterol, triglycerides, and LDL versus HDL of mice treated with olive oil and N. sativa oil were all significantly below the respective control values as a result of reducing lipid profiles and increasing HDL levels (Table 2).

Table 2. Ratios of cholesterol, triglycerides, and low-density lipoprotein cholesterol versus high-density lipoprotein cholesterol in male mice orally treated with olive oil and Nigella sativa oil at 0.4 ml/kg/day for 60 consecutive days.

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Discussion

The present experimental results show that treating mice with olive oil and N. sativa oil for 60 days significantly improved their serum lipid profiles compared to controls under normal feeding conditions. However, these improvements remained within the physiological ranges because the baseline lipid levels were already normal. Specifically, both interventions markedly reduced the serum levels of total cholesterol, triglycerides, LDL, and VLDL while simultaneously increasing the HDL levels. These findings highlight the potential protective effects of these natural oils against lipid disorders and align with earlier studies in animals with hyperlipidemia, where both olive oil and N. sativa seeds showed positive effects (Kafi, 2014; Katsarou et al., 2016; Aboul-Mahasen et al., 2019; Albrahim et al., 2022). Similar trends were observed in human studies, supporting the idea that these natural oils can lower serum lipid levels (Asgary et al., 2015; Buckland and Gonzalez, 2015; Montserrat-de la Paz et al., 2016; Ferro et al., 2023; Jabbarzadeh-Ganjeh et al., 2023). This improvement in the lipid profile is crucial because high cholesterol, triglycerides, and LDL/VLDL levels significantly contribute to the risk of cardiovascular disease (Alloubani et al., 2021; Wright and Subramanian, 2024; Wierzbicki, 2025). The increase in HDL is advantageous because it aids in transporting cholesterol from body tissues to the liver for elimination, thus protecting against atherosclerosis and other lipid disorders (Tarmoos and Kafi, 2019; Bhale et al., 2024). Traditional medicine using these oils also supports these notions.

A key observation in this study is the significant reduction in the ratios of cholesterol, triglycerides, and LDL to HDL in the treated groups relative to the controls. These ratios are clinically useful indicators of potential cardiovascular risk, as lower values generally reflect a favorable lipid profile and reduced risk of atherogenesis (Asgary et al., 2015; Alloubani et al., 2021; Wright and Subramanian, 2024; Wierzbicki, 2025). No differences were observed between olive oil and N. sativa oil despite their distinct bioactive compounds. This could be attributed to the fact that both oils target the same changes in the lipid profile, but they do so through different mechanisms. The mechanisms of action of olive and N. sativa oils may differ. Olive oil, particularly extra virgin olive oil, contains high levels of monounsaturated fatty acids and polyphenols, which increase HDL levels and reduce LDL oxidation (Buckland and Gonzalez, 2015; Katsarou et al., 2016; Aboul-Mahasen et al., 2019; Farias-Pereira et al., 2023). On the other hand, N. sativa oil contains thymoquinone and other bioactive compounds that may be a factor in lipid-lowering effects through mechanisms that involve prevention of lipid peroxidation and inflammation, as well as enhancing insulin action (Ali and Blunden, 2003; Ahmad and Beg, 2013; Asgary et al., 2015; Aboul-Mahasen et al., 2019). It may also decrease cholesterol by reducing its absorption in the intestines, lowering its production in the liver, and increasing LDL receptors (Asgary et al., 2015). However, some studies suggest that N. sativa may not raise HDL levels as effectively as olive oil, and in some instances, it might even lower HDL levels (Asgary et al., 2015; Aboul-Mahasen et al., 2019). Additional studies are needed to comparatively explore these effects following long-term investigations into molecular mechanisms. The potential interaction between these two oils when administered simultaneously remains unknown, and the possibility of a synergistic effect between them must be investigated. In clinical settings, however, N. sativa has shown effectiveness when used along with standard drugs for lowering hyperlipidemia and managing diabetes (Aboul-Mahasen et al., 2019). In addition to improving blood lipid levels, both oils have additional metabolic and molecular benefits, such as better glucose control, reduced serum amylase, enhanced pancreatic tissue healing, reduced cellular toxicity, and higher insulin production (Kafi, 2014; Asgary et al., 2015; Kafi, 2015; Aboul-Mahasen et al., 2019; Tarmoos and Kafi, 2019). Both olive oil and N. sativa oil are readily available locally, providing an additional nutritional benefit to the public.

Limitations of the study

The small number of mice in each group is one of the study’s limitations. However, this is in alignment with the ARRIVE guidelines and the concept of 3Rs in reducing the number of animals in the experiments. This study is not mechanistic (e.g., oxidative stress modulation) and does not consider sex or genetic factors that influence poor lipid profiles. However, future studies should investigate these aspects, including pharmacogenetics and gender, along with other relevant factors.

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Conclusion

In conclusion, this study confirms that olive oil and N. sativa oil can significantly improve blood lipid levels in normolipidemic mice, particularly by lowering atherogenic lipids and increasing HDL. These changes result in favorable cholesterol, triglyceride, and low-density lipoprotein/high-density lipoprotein ratios, which play an important role in lowering cardiovascular disease risk and highlight the potential therapeutic and possibly protective benefits of these oils in managing lipid disorders.

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Acknowledgment

The author would like to thank the Animal House at the College of Veterinary, University of Baghdad, Iraq, for providing the mice and facilities.

Conflict of interest

The author declares that there are no conflicts of interest.

Authors’ contributions

The author conceptualized the study, conducted the experiments, analyzed the data, drafted the manuscript, and finally reviewed it for publication.

Funding

None to declare.

Data availability

Data will be available from the author upon reasonable request.

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