Bergamot Extracts as Anti-Aging Agents for Red Blood Cells: A Comprehensive Exploration


Aging is an inevitable biological process that takes its toll on the physiological functions of all body organs over time. It is characterized by a progressive and non-reversible decline in the performance of these vital systems.

This decline can be attributed to the accumulation of damage and an increase in oxidative stress within the body. Over the years, several theories have attempted to explain the underlying mechanisms of aging, one of which is the free radical theory of aging proposed by Denham Harman in 1972.

This theory suggests that the accumulation of reactive species, or free radicals, is responsible for the oxidation of biological macromolecules and subsequent cell injury, leading to the alterations observed in cell functions during natural aging.

The Role of Reactive Oxygen Species

Aerobic cell metabolism, which relies on the presence of oxygen as the final electron acceptor in oxidation reactions, results in the generation of reactive oxygen species (ROS) as byproducts.

Red blood cells (RBCs) play a crucial role in transporting respiratory gases, specifically oxygen, from the lungs to the body’s tissues and vice versa. In the bloodstream, RBCs are continually exposed to both endogenous and exogenous reactive species that can damage their structure and compromise their functionality.

Unchecked reactive species can react with the lipids and proteins in RBC plasma membranes, leading to lipid peroxidation and protein oxidation or fragmentation. Furthermore, the accumulation of reactive species triggers glycation reactions, resulting in the production of advanced glycation end products (AGEs). This process can lead to changes in the rheological properties of RBCs.

Effects of Oxidative Stress on RBC Aging

Oxidative stress-related aging in RBCs is associated with several significant changes, including a decrease in cell volume and hemoglobin content, along with an increase in cell density. These alterations are closely linked to the loss of cholesterol and phospholipids, resulting in a reduction in surface area, which is indicative of a loss of membrane lipids and protein constituents.

This reduction in surface area can be attributed to membrane blebbing and vesiculation, processes that are accelerated in aged RBCs. These changes limit the RBCs’ ability to maintain their highly deformable biconcave shape, which is necessary for them to pass through narrow capillaries. Consequently, these alterations contribute to the premature removal of RBCs from circulation.

Role of Band 3 Protein

One of the primary goals of RBC redox regulation is to protect their main plasma membrane protein, band 3 (SLC4A1/AE1). Band 3 is a transmembrane protein that serves multiple functions, including maintaining anion homeostasis, connecting plasma membrane lipids to cytoskeletal proteins, and acting as a docking site for various structural proteins and glycolytic enzymes. Additionally, band 3 can bind to cytosolic proteins such as hemoglobin. Hemoglobin, aside from its role in oxygen transport, serves as an oxygen sensor and plays a role in the regulation of RBC metabolism.

The balance between glycolysis and the pentose phosphate pathway (PPP) is crucial for RBCs.

When oxidant stress is high, glycolytic enzymes are bound to band 3, leading to glucose oxidation through the PPP to generate NADPH, which fuels endogenous antioxidant systems. Conversely, when oxidant stress is low, deoxygenated hemoglobin binds to band 3, promoting the release of glycolytic enzymes and favoring energy generation through glycolysis.

This balance can be disrupted by oxidative stress or RBC aging, leading to a loss of metabolic plasticity and potentially contributing to their premature removal from circulation.

Protective Mechanisms in RBCs

To counteract the detrimental effects of oxidative stress, RBCs are equipped with a robust cytosolic antioxidant system, comprising both non-enzymatic and enzymatic antioxidants. These antioxidants protect not only RBCs themselves but also other cells and tissues in the body. In this context, natural secondary metabolites, such as polyphenol-rich extracts with antioxidant properties, have shown promise as a means to support the integrated antioxidant system. One such source of these beneficial compounds is the bergamot, particularly its flavonoid fraction.

Bergamot: A Potential Anti-Aging Solution

Bergamot, a small tree belonging to the Rutaceae family, thrives in a specific microclimate in the region of Calabria, Italy. While the essential oil of bergamot has been extensively utilized in the cosmetic and food industries, recent attention has turned to the potential health benefits of by-products such as pulp and juice. These benefits include cholesterol reduction, antioxidant properties, and anti-inflammatory effects, all of which could be harnessed as nutraceuticals. However, scientific studies exploring the anti-aging properties of bergamot extracts on human RBCs have been lacking.

The Experimental Model of Natural Aging

To investigate the potential anti-aging effects of bergamot peel and juice extracts from the Femminello cultivar, a model of natural aging was employed. This model utilizes long-term exposure to high doses of D-Galactose (D-Gal) to simulate the aging process in the laboratory setting.


The aging process, driven by the accumulation of oxidative stress and damage to biological macromolecules, plays a significant role in the development of age-related diseases. Natural antioxidant compounds have gained increasing attention due to their potential in retarding or even reversing the course of such diseases.

These antioxidants can compete with substrates sensitive to oxidation, thus inhibiting or delaying reactions between reactive species and biological macromolecules. Excessive production of reactive species can lead to cellular damage and the onset of chronic diseases. Dietary intake of natural antioxidants can boost the endogenous antioxidant system, aiding in the prevention of pathological conditions.

In this study, we explored the potential anti-aging effects of bergamot peel and juice extracts on oxidative stress-induced aging, focusing on human red blood cells (RBCs). We employed a cell-based model involving prolonged exposure to D-Galactose (D-Gal) to simulate natural aging. Bergamot extracts, including both peel and juice, were tested across a range of concentrations and incubation times to evaluate their effects on RBCs and exclude potential damage caused by direct exposure to the extracts.

Notably, higher concentrations of peel or juice extract, particularly when combined with prolonged incubation times, were found to induce hemolysis and exhibit a pro-oxidant effect. This emphasizes the importance of careful assessment of concentration and incubation time when testing potential antioxidant compounds in cell-based assays.

For subsequent analyses, we selected a 15-minute pre-treatment with 5 μg/mL of either peel or juice extract to estimate the antioxidant capacity. We measured levels of reactive oxygen species (ROS), thiobarbituric acid reactive substances (TBARS), and total content of sulfhydryl groups in RBCs that had been incubated for 24 hours with 100 mM D-Gal.

RBCs are particularly vulnerable to ROS-induced damage due to their high polyunsaturated fatty acid content and the presence of iron (Fe2+)-rich hemoglobin, which acts as a catalyst in redox reactions and lipid peroxidation. Our results showed that a 15-minute pre-exposure of RBCs to 5 μg/mL peel or juice extract effectively prevented ROS production induced by D-Gal.

Moreover, these extracts provided protection against both lipid and protein components of the RBC plasma membrane from oxidative damage. This indicates that the bioactive molecules in bergamot extracts have scavenging properties, neutralizing reactive species and free radicals to prevent their harmful impact on biological macromolecules.

Aging also affects the plasma membrane transport systems of RBCs, especially the anion exchange mediated by the band 3 protein. The rate of sulfate (SO42−) uptake, used as a measure of anion exchange, was accelerated in RBCs exposed to D-Gal. This phenomenon reflects the impact of oxidative stress on band 3 in human RBCs, a finding that aligns with previous research.

Notably, a 15-minute pre-treatment of RBCs with 5 μg/mL peel or juice extract was able to partially or fully restore the rate constant for SO42− uptake. This suggests that these extracts have a protective role against oxidative stress-induced functional changes in RBCs, which are essential for maintaining homeostasis.

The interactions between band 3 and hemoglobin are critical for RBC function. Exposure to D-Gal caused the shedding of protein-containing vesicles, impacting both the expression and distribution of band 3. Despite this, pre-treatment with 5 μg/mL of peel or juice extract was found to restore band 3 expression levels. These extracts play a role in preventing membrane shedding, glycated hemoglobin formation, and structural instability, all of which are characteristics of aging RBCs.

Oxidative stress can exhaust the endogenous antioxidant defense system. In this study, the activities of catalase (CAT) and superoxide dismutase (SOD) were measured.

Both CAT and SOD play vital roles in RBCs. In response to D-Gal treatment, the activities of these enzymes were significantly increased. However, this increase was insufficient to compensate for the elevated free radicals and subsequent lipid peroxidation and protein oxidation. Furthermore, the GSH/GSSG ratio, a marker of redox balance, was reduced by D-Gal treatment. Pre-incubation with peel and juice extract was effective in preventing the upregulation of CAT and SOD activities and restoring the GSH/GSSG ratio, indicating that these extracts may work synergistically with the endogenous antioxidant system to counteract oxidative stress and preserve RBC integrity.

In summary, this study sheds light on the potential anti-aging properties of bergamot peel and juice extracts, emphasizing their protective role against oxidative stress-induced aging in human RBCs. These extracts prevent oxidative damage to both the lipid and protein components of the RBC plasma membrane, maintain anion exchange, and restore the redox balance. The findings presented here provide valuable insights into the use of natural antioxidant compounds in mitigating the effects of aging, ultimately contributing to overall health and well-being. Further research in this area may uncover new strategies to combat age-related diseases and enhance longevity.

In deep…..

Bergamot Extracts as Anti-Aging Agents for Red Blood Cells

Red blood cells (RBCs) are essential for carrying oxygen throughout the body. As we age, RBCs become less efficient at transporting oxygen, which can lead to a number of health problems, including fatigue, shortness of breath, and cognitive decline.

Bergamot extracts have been shown to have anti-aging properties in both in vitro and in vivo studies. Bergamot is a citrus fruit native to Italy and is known for its unique fragrance and flavor. It is also a good source of flavonoids, which are powerful antioxidants.

How Bergamot Extracts Protect RBCs

Bergamot extracts protect RBCs from aging in a number of ways, including:

  • Reducing oxidative stress: Oxidative stress is a major contributor to RBC aging. Bergamot extracts contain flavonoids, which are powerful antioxidants that can neutralize free radicals and protect RBCs from oxidative damage.
  • Improving membrane fluidity: As RBCs age, their membranes become less fluid. This can make it difficult for RBCs to transport oxygen efficiently. Bergamot extracts have been shown to improve membrane fluidity in RBCs, which can help to improve their oxygen-carrying capacity.
  • Increasing antioxidant enzyme activity: RBCs contain a number of antioxidant enzymes that help to protect them from oxidative damage. Bergamot extracts have been shown to increase the activity of these enzymes, which can help to further protect RBCs from aging.

How to Use Bergamot Extracts to Protect RBCs

There are a few different ways to use bergamot extracts to protect RBCs:

  • Supplements: Bergamot extracts are available in supplement form, typically in capsule or powder form. The recommended dosage for bergamot extract supplements varies depending on the product. It is important to follow the dosage instructions on the product label.
  • Tea: Bergamot tea can also be used to protect RBCs. Bergamot tea is made by steeping bergamot leaves or flowers in hot water. To make bergamot tea, add 1-2 teaspoons of dried bergamot leaves or flowers to a cup of hot water. Steep for 5-10 minutes before drinking.
  • Essential oil: Bergamot essential oil can also be used to protect RBCs. Bergamot essential oil is very concentrated, so it is important to dilute it before using it. To dilute bergamot essential oil, add 1-2 drops of oil to a carrier oil, such as jojoba oil or almond oil. The diluted oil can then be applied to the skin or used in aromatherapy.

When to Use Bergamot Extracts

Bergamot extracts can be used at any age to help protect RBCs from aging. However, they may be particularly beneficial for older adults, who are at increased risk of RBC aging and its associated health problems.

Safety Considerations

Bergamot extracts are generally safe for most people. However, it is important to talk to your doctor before using bergamot extracts, especially if you have any underlying health conditions.

Bergamot essential oil is photosensitive, so it should not be applied to the skin before going out in the sun. It is also important to do a patch test before using bergamot essential oil or extract on the skin to make sure that you are not allergic to it.


Bergamot extracts are a safe and effective way to protect RBCs from aging. Bergamot extracts can be used in a variety of ways, including supplements, tea, and essential oil. If you are interested in using bergamot extracts to protect your RBCs, talk to your doctor first to make sure that they are right for you.

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