The Nrf2 Pathway and Nutrition: What You Need to Know

Genes Mentioned

• GSTP1
• NQO1

Did you know that your body makes some of its own antioxidants? That’s right, antioxidants aren’t exclusively to be found in a supplement bottle, we produce our own “endogenous” antioxidants to help protect our cells from damage. Oxidative stress damages cells, so as a protection mechanism to make sure levels of cellular stress don’t get too high, the body produces enzymes such as superoxide dismutase 2 (SOD2), or specific beneficial nutrients such as glutathione. These direct acting enzymes and nutrients are great at fighting the fire of oxidative stress, which is why the body makes more of them when it feels under attack. When oxidative stress levels get too high, the Nrf2 pathway is activated. The Nrf2 pathway is the system the body turns on when it wants to make its “in-house antioxidants” and there are nutrients we can turn to to activate this pathway as well. Nutrients known to activate Nrf2 include:

• sulforaphane from broccoli seed extract;
• curcumin from the turmeric plant;
• and carotenoids such as zeaxanthin and lutein.

Before we dive into the importance of the Nrf2 pathway, and how to activate it, we’ll go over a little primer on what happens when our bodies generate energy at the cellular level.

Cellular energy generation produces free radicals

Most compounds and molecules in our body exist in a relatively stable state, this is why we need enzymes (think of them as molecular power tools) to break them down or turn them into other more useful molecules. This is how the body produces energy. However, the byproduct of energy metabolism are damaging free radicals.

Free radicals are unstable

These free radicals are chemically unstable, and highly reactive. Two common examples are O₂^– sometimes called superoxide and H₂O₂ or hydrogen peroxide. O₂ is the normal stable form of oxygen, however when involved in some reactions in the body O₂^– is formed. That little ^– sign means that an extra electron has been added. Superoxide is therefore highly reactive as it is trying to find a way to lose that extra electron, and the only way to do that is to transfer it to another molecule or compound.

Antioxidants clean up free radicals

This is where antioxidants step in, it’s their function to mop up these highly reactive species making sure they don’t interact with any of the important molecules in a cell. This mopping up occurs constantly in the body, however, when antioxidant function in the body becomes overwhelmed by free radicals, oxidative stress can occur. ¹

Free radicals are a necessary part of energy production

It is important to understand that free radicals in themselves are not bad, their formation is part and parcel of cellular energy generation, and they serve a vital role in other functions such as bacterial killing.² So your aim should never be to remove free radicals, but rather ensure your system is in balance.

Oxidative stress and disease

So, the goal is not to stop the production of free radicals, but rather to stay in balance so the body has adequate opportunity to neutralize these compounds. However chronic oxidative stress, where levels of stress overwhelm the body’s systems for cleaning it up, is associated with several diseases including Parkinson’s disease, Alzheimer’s disease, cardiovascular disease and several cancers.³⁴⁵⁶ The exact impact is hard to pin down due to the long time scales involved in the development of these diseases, but it is an area of very active research. Other major areas of interest are dietary and environmental exposures. At a base level things like alcohol and tobacco or a meat rich diet can increase oxidative stress. However, these are relatively easy to avoid and if you’re reading this blog chances are you’re already making positive health choices. There are other factors however which are more difficult to avoid such as air pollution, excessive exposure to pesticides, herbicides, or heavy metals.⁷⁸⁹¹⁰ In these cases chronic oxidative stress can be generated, leaving existing anti-oxidant defenses overwhelmed.

Fighting chronic oxidative stress

So we know that endogenous antioxidants, such as glutathione, are great at mopping up free radicals on a one to one basis, and that enzymes such as the SOD or glutathione peroxidase (GPX) enzyme families can rapidly reduce oxidative stress. But what happens in a chronic system where these defenses have been overwhelmed?

The Nrf2 Pathway and antioxidant activity

Nuclear factor erythroid 2 [NF-E2]-related factor 2, known by the much easier to remember acronym Nrf2 is a transcription factor which is a major regulator of cell protection, including antioxidant, responses.¹¹ Transcription factors are a class of protein which bind to DNA and induce the expression of particular genes, in the case of Nrf2, the pathway stimulates the production of potent antioxidants such as: NAD(P)H:quinone oxidoreductase 1 (NQO1) and glutathione S-transferases (GSTs), and many others, as shown in the table below. ^{12 13} Table 1 – Genes activated by NRF2. Adapted from: Baird, L. and Dinkova-Kostova, A.T. Arch. Toxicol. 85,241-272 (2011)

Nrf2 in health

In health, Nrf2 exists in the cytosol of a cell, or the region outside of the nucleus where it cannot interact with DNA. It is held here by another protein called Keap1, which prevents it moving into the nucleus. ¹⁴

Inflammation stimulates Nrf2 activation

Keap1 contains several sensors for reactive oxygen species, along with receptor regions for other cell proteins associated with cell stress.¹⁵ If these receptors are activated then Nrf2 is released, and can pass into the nucleus.¹⁶ Once within the nucleus Nrf2 binds to a region of DNA known as an “Antioxidant Response Element” or ARE.¹⁷ These ARE elements are closely associated with the genes for NQO1 and the GSTs (and many other genes) as discussed above, and when Nrf2 binds it induces the production of active proteins from these genes. Both of these proteins exhibit potent antioxidant capacity and can rapidly clear oxidative stress. The important role for Nrf2 has been shown by several studies with a particular focus on animal models. Mice which have been genetically engineered to lack Nrf2 protein are considerably more sensitive to chemical carcinogens, and also chemical toxins leading to increased inflammation in the lung and brain, all major markers of oxidative stress.

Foods and supplements that activate Nrf2

While oxidative stress alone is able to activate Nrf2, its effect is greatly enhanced by the presence of certain chemical compounds. The simplest of these compounds were identified as 1,4-benzenediol (hydroquinone), tert-butylhydroquinone, and 1,2-benzendiol (catechol), and more complex examples include the isothiocyanate sulforaphane from broccoli seed extract, curcumin from the turmeric plant and carotenoids such as zeaxanthin and lutein. All are complex in name and structure, but they share a common function in that they support increased and prolonged expression of Nrf2 gene targets.^{18 19}

Issues with Nrf2 activation

Whilst Nrf2 activation is undoubtedly beneficial in the vast majority of cases, there are some instances where it may prove harmful. Once again using a mouse model, this time lacking Keap1, meaning that Nrf2 would be constantly activated, researchers demonstrated that severe health effects would develop.²⁰

Nrf2 and cancer risk

Additionally, mutations in the Nrf2 gene in human studies have been associated with an increased risk of cancer, with the authors proposing that constance Nrf2 activity, prevents anti-cancer targeted oxidative stress, allowing cancer cells to survive.²¹ It is unlikely that dietary supplementation would ever induce such strong effects but it’s a useful example of how oxidative stress isn’t always bad. Importantly there are no currently reported SNPs in Nrf2 which impact on its function. A lack of variation in genes like this typically points to how important they are to life, however an alternative option is that Nrf2 has not been widely investigated.

Take-home message on Nrf2

The take home message in this instance is quite simple. Nrf2 is a potent modulator of antioxidant response and can rapidly target oxidative stressors. While Nrf2 responds to oxidative stress directly, certain key nutrients can improve this effect, the exact source of these nutrients varies widely but dark green and leafy vegetables (think spinach, kale and broccoli, especially the seed extract), or reddy vegetables and spices (saffron, tumeric or paprika) are a particularly rich source. Importantly, these foods are also typically rich in direct antioxidants and display numerous other health benefits, so they are a great way to improve overall health.