Hypercholesterolemia or dyslipidemia occurs when low-density lipoprotein (LDL) rises and accumulates fat in the arteries (atherosclerosis) and cardiovascular diseases, increasing the risk of heart attacks and contributing to more deaths (El Rabey et al., 2018; Guo et al., 2022; Li et al., 2023). It is a severe health issue that impacts the body’s oxidative stress, rendering it vulnerable to a variety of illnesses, including Alzheimer’s, osteoporosis, diabetes, high blood pressure, cancer, obesity, and aging (Al-Sieni et al., 2020; Jomova, et al., 2023; Li et al., 2023).

In addition, hyperlipidemia induces oxidative stress through the buildup and oxidation of low-density lipoproteins (LDL) cholesterol in artery walls, which leads to inflammation and ultimately artery narrowing that, in turn, raises the risk of heart attacks, strokes, and cardiovascular diseases (Zmysłowski and Szterk, 2017; Prasad and Mishra, 2022). In addition, the World Health Organization (WHO) estimates that approximately 39% of people worldwide have high cholesterol (Sawhney and Madan, 2024). The frequency of familial hypercholesterolemia (FH) in the general population is estimated to be between 1 in 250 to 1 in 311 (Vaezi and Amini, 2025). Regional variations exist, with founder effects leading to a higher incidence in specific populations in the Middle East, Quebec, and South Africa (Vaezi and Amini, 2025; Sawhney and Madan, 2024).

Chemical-based medicines may cause adverse reactions, such as liver problems, nausea, drowsiness, diarrhea, or allergic reactions, known as an adverse drug reaction (ADR) (Kommu et al., 2025; Coleman and Pontefract, 2016). To avoid the negative effects of chemical medicine, people all over the world prefer to use herbal medicine to treat and prevent hypercholesterolemia. This has led scientists to investigate the benefits of employing herbal remedies and natural products to reduce hypercholesterolemia, lessen the risk of cardiovascular disorders, and shield the heart from heart attacks (El Rabey et al., 2018; Al-Sieni et al., 2020; Alshareef et al., 2024).

The safflower (Carthamus tinctorius L.; Family Asteraceae) is a highly branched annual herbaceous plant grown for its culinary and medicinal properties (Delshad et al., 2018; Adamska and Biernacka, 2021). Its yellowish-red blossoms are used for many medical purposes, and its seeds are crushed to yield edible oil. Because of its analgesic and antipyretic properties, it has also been used in traditional medicine to alleviate the symptoms of menstrual discomfort. It was also used in food coloring and flavoring as well as other medicinal applications such as a purgative, an antidote for poisoning, and as a remedy for postpartum hemorrhage, trauma, joint discomfort, and osteoporosis (Delshad et al., 2018).

Worldwide, safflower seeds have long been used to prevent osteoporosis and encourage bone growth (Che et al., 2016). Carthamin, safflower yellow, safflamin C, isocarthamidin, hydroxysafflor yellow A, Carthamidin, safflor yellow A, luteolin caryophyllene, 1-acetoxytetralin, heneicosane, and p-allyltoluene are the primary active ingredients of safflower flowers (Delshad et al., 2018). According to Adamska and Biernacka (2021), the primary ingredient in safflower flower extract, hydroxysafflor (HSYA), exhibits anti-inflammatory, antibacterial, and antioxidant properties, and has been shown to combat obesity in rats and mice.

The FDA-approved medication atorvastatin, often known as Lipitor, is a synthetic drug of the statin class used to treat high blood pressure and heart issues by preventing cardiovascular illnesses in patients with dyslipidemia and cardiac risk factors (Livingstone et al., 2016; Abd-Elmonsif and Gamal, 2025).

Vitamin D is a micronutrient, sunlight-dependent vitamin that maintains a healthy skeleton and guards against liver fibrosis and is considered vital for numerous metabolic processes (Che et al., 2016; Keane et al., 2018). Rickets, osteomalacia, cancer, and cardiovascular disease all occur due to a lack of vitamin D, which is either produced endogenously as vitamin D3 (cholecalciferol) or acquired through diet (ergocalciferol) (Zhang et al., 2017; Deepika et al., 2025). Numerous diseases, including inflammation, carcinogenesis, endocrinopathies, autoimmunity, type 1 and type 2 diabetes mellitus, Hashimoto’s or Graves’ thyroiditis, Cushing’s disease, hyperaldosteronism, adrenocortical tumors, and the adrenal gland, have been linked to vitamin D, according to both in vitro and in vivo research (Al Refaie et al., 2022).

As a chemical messenger that serves as a central nervous system neurotransmitter to convey nerve impulses within nerve, muscle, or gland cells, adrenaline, also known as epinephrine, is generated by the adrenal glands and is “a part of the emergency response system to the fight-or-flight response.” Costa and Schoenbaum, 2022). Sugar consumption and hypercholesterolemia cause metabolic dysfunction as well as other mental and neurological conditions such as anxiety and depression (Fulton et al., 2022).

This study was focused at clarifying the probable hypolipidemic effect of atorvastatin and safflower in male rats with induced hypercholesterolemia. Some other therapeutic activities of both materials were also studied.

This study evaluated the therapeutic advantages of C. tinctorius aqueous extract and atorvastatin in male rats with induced hypercholesterolemia. This investigation utilized the aqueous extract of safflower flowers, a prevalent food ingredient, and the trial was limited to 4 weeks to mitigate the risk of toxicity associated with prolonged safflower administration (Adamska and Biernacka, 2021; Alshareef et al., 2024).

Administration of a fat-rich food to the experimental animals resulted in hypercholesterolemia in the positive control group, as evidenced by increased triglycerides, total cholesterol, LDL, and vLDL, alongside a reduction in HDL (El Rabey et al., 2018; Alshareef et al., 2024). The administration of safflower and atorvastatin significantly restored the lipid profile of the hypercholesterolemic rats in G3 and G4, respectively. C. tinctorius has various health advantages, including antioxidant, antidiabetic, anti-inflammatory, and analgesic properties, in addition to its nutritional benefits (Che et al., 2016). These significant therapeutic benefits can be attributed to its higher content of polyphenols, such as quercetin, chlorogenic acid, and HSYA’s (Adamska and Biernacka, 2021). The polyphenols of safflower, such as quercetin and chlorogenic acid, and HSYA’s enhance the production and activity of hormone-sensitive lipase (HSL) while inhibiting adipocyte proliferation (Sandoval et al., 2020). Conversely, atorvastatin is an approved drug for lowering cholesterol levels and preventing cardiovascular diseases (Livingstone et al., 2016; Abd-Elmonsif and Gamal, 2025). It reduces cholesterol by inhibiting 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, thereby preventing the conversion of HMG-CoA to mevalonate, which diminishes cholesterol synthesis in the liver and enhances the quantity of LDL receptors on hepatic cells (McIver and Siddique, 2025; Dagli-Hernandez et al., 2022; Abd-Elmonsif and Gamal, 2025).

The induction of hypercholesterolemia in the positive control group markedly impacted liver function, elevating liver enzymes and bilirubin levels while diminishing total protein (Alshareef et al., 2024). Furthermore, these modified liver functions resulted from the direct manifestation of the histopathological damage caused by hypercholesterolemia on the liver tissues (Al-Sieni et al., 2020). The hepatoprotective effect of safflower in the treatment group may be attributed to the free radical scavenging properties of its polyphenolic compounds (Ruyvaran et al., 2022; Alshareef et al., 2024). Moreover, atorvastatin’s protective effects may stem from its essential function in inhibiting fat buildup, hence enhancing the functionality of other organs and mitigating cardiovascular illnesses and dyslipidemia (Livingstone et al., 2016; Abd-Elmonsif and Gamal, 2025). Safflower also protected against liver damage and hepatic steatosis in type 2 diabetes mellitus via Nrf2-dependent pathways, attributable to its antioxidant, hypoglycemic, hypolipidemic, and anti-inflammatory properties (Alshareef et al., 2024).

The induction of hypercholesterolemia resulted in increased lipid peroxidation, as indicated by elevated MDA levels and reduced activity of the antioxidant enzymes catalase, SOD, and GSH, in comparison to the negative control group. Conversely, treatment with safflower and atorvastatin improved these disrupted oxidative stress parameters. Furthermore, the hepatoprotective properties of safflower may be attributed to polyphenols such as quercetin, chlorogenic acid, and HSYA’s ability to augment the synthesis of hepatic antioxidative enzymes that regulate obesity. Additionally, the administration of safflower extract, which comprises both HSYA and safflower yellow (also known as carthamine yellow), elevates the mRNA levels of antioxidative enzymes and enhances the superoxide dismutase (SOD) activity in liver homogenates (Yan et al., 2020). HSYA addresses obesity by modifying intestinal microbiota, enhancing particular digestive bacterial populations in the colon, and optimizing digestive tract function and systemic metabolism (Adamska and Biernacka, 2021).

The elevation of heart function enzyme activity (troponin and CK) in the hypercholesterolemic positive control group is corroborated by histological analyses that indicated damaged cardiac tissues and oxidative stress markers (El Rabey et al., 2018; Al-Sieni et al., 2020). The notable enhancement in cardiac function metrics following treatment with safflower and atorvastatin may be attributed to their hypolipidemic and antioxidant properties, which alleviated oxidative stress (Yan et al., 2020; Abd-Elmonsif and Gamal, 2025). Recent studies have identified hyperlipidemia as a risk factor for stroke in patients with coronary vascular diseases (CVD), prompting ongoing research aimed at mitigating this risk factor to reduce the incidence of stroke and myocardial infarction (MI) (Kim et al., 2017; Olamoyegun et al., 2016; Alloubani et al., 2021).

The current findings indicated a notable reduction in vitamin D and calcium levels due to induced hypercholesterolemia in the positive control group, while treatment with safflower and atorvastatin resulted in an elevation of these levels. Calcium and vitamin D levels exhibit a positive correlation; therefore, calcium is essential for vitamin D production. Recent research indicates that hypercholesterolemia is associated with a deficiency in vitamin D levels and vice versa (Dibaba, 2019; Han et al., 2021). The elevation of calcium levels is associated with the enhancement of vitamin D to optimal levels; thus, vitamin D is integral to calcium homeostasis at these optimal levels (Mattioli et al., 2025). Furthermore, Che et al. (2016) indicated that safflower seeds exhibit protective effects against bone loss induced by estrogen shortage through the stimulatory action of polyphenolic chemicals on osteoblast growth. Vitamin D facilitates the development of healthy calcified vertebrate skeletons, whereas its receptor in calcium-regulating organs modulates calcium metabolism and enhances bone health (Izzo et al., 2024). In addition, liver illness is associated with a deficit in vitamin D (Elangovan et al., 2017). Vitamin D supplementation safeguards liver health from fibrosis, and its production is contingent upon the liver’s health and functionality (Keane et al., 2018).

The induced hypercholesterolemia markedly elevated adrenaline levels in the present investigation. This finding is consistent with earlier research linking hypercholesterolemia to elevated adrenaline levels (Maduka et al., 2015; Fulton et al., 2022). Treatment of hypercholesterolemic rats with safflower and atorvastatin enhanced adrenaline levels, practically returning them to normal as observed in the negative control group. This enhancement is associated with the hypolipidemic effects of safflower and atorvastatin, which benefit the liver. Adrenaline is a catecholamine, along with dopamine and norepinephrine, that significantly contributes to the progression of liver disorders (Lelou et al., 2022). The preventive impact of safflower may be attributed to its elevated levels of the polyphenol’s quercetin and chlorogenic acid, recognized for their superior antioxidant, antidiabetic, and hypolipidemic properties (Jiang et al., 2019; El Rabey et al., 2018; Liu et al., 2022). The association of vitamin D with hypercholesterolemia and the adrenal gland is attributed to their shared genetic pathway (Al Refaie et al., 2022).

The induced hypercholesterolemia also caused hepatic and cardiac histological alterations in the positive control group relative to the negative control (El Rabey et al., 2018; Al-Sieni et al., 2020). These altered changes in the hepatic and cardiac tissues caused an elevation in the liver and heart markers in the positive control group (Lelou et al., 2022; Ognik et al., 2021). Administration of safflower and atorvastatin to hypercholesterolemic rats restored the histology of liver and heart, approaching the normal state and protecting against histological alterations (Lelou et al., 2022; Liu et al., 2022).

The total body weight was gradually increased with increasing hypercholesterolemia (Al-Sieni et al., 2020; El Rabey et al., 2020; Liu et al., 2022), while treating these hypercholesterolemic rats with safflower and atorvastatin significantly ceased this weight gain. The quercetin (an Omega-3 fatty acid) and chlorogenic acid, which are naturally occurring phenolic compounds, represent the major constituents of safflower, which play a significant role in improving lipid metabolism and thus decreasing weight gain compared to the positive control group (Jiang et al., 2019; Tahir et al., 2023). Similarly, atorvastatin treated the hypercholesterolemia and thus attenuated the increased body weight gain (Livingstone et al., 2016; Abd-Elmonsif and Gamal, 2025).

Atorvastatin is an effective cholesterol-lowering medication recommended by the FDA (Livingstone et al., 2016; Abd-Elmonsif and Gamal, 2025). In this study, we observed that treatment with safflower increased antioxidant enzyme activity, calcium, and vitamin D, and decreased total cholesterol, LDL, triglycerides, lipid peroxidation, liver function enzymes, adrenaline, troponin, and CK levels (Fulton et al., 2022; Alshareef et al., 2024; Deepika et al., 2025).

Safflower extract has been used in folk medicine for short periods and in specific doses to avoid toxicity resulting from large doses. It is used to treat many diseases such as hyperlipidemia, diabetes, and many symptoms associated with anxiety, stress, and fear. Hence, the idea for this study arose, and we hope that the results of this study will be used to develop pharmaceutical formulations of safflower to treat these diseases and alleviate pain. The dosage of safflower extract may need to be elevated to achieve enhanced protective efficacy. Additional research is necessary to investigate the function of each component in improving health and elucidating its protective effects on different organs. In addition, this study was conducted on rats, despite they do not accurately model human lipoprotein profiles or atherosclerosis progression, because rodents are considered ideal for atherosclerosis research because of their fast breeding, low cost of production, and short time needed for developing atherosclerosis (Daniels et al., 2018; Zhao et al., 2020).

Safflower and atorvastatin exhibited numerous health advantages. They elevated antioxidant and vitamin D levels and reduced hyperlipidemia, lipid peroxidation, CK, troponin, and adrenaline. They further safeguarded the liver and heart of the generated hypercholesterolemic animals. They also function as anxiolytics by reducing adrenaline levels. Atorvastatin has shown superior efficacy compared to safflower in safeguarding against hypercholesterolemia and its associated consequences.