Polyphenols, Hormesis and Disease: Part I

What are Polyphenols?

Polyphenols are a diverse class of molecules containing multiple phenol rings. They are synthesized in large amounts by plants, certain fungi and a few animals, and serve many purposes, including defense against predators/infections, defense against sunlight damage and chemical oxidation, and coloration. The color of many fruits and vegetables, such as blueberries, eggplants, red potatoes and apples comes from polyphenols. Some familiar classes of polyphenols in the diet-health literature are flavonoids, isoflavonoids, anthocyanidins, and lignins.

The Case Against Polyphenols

Mainstream diet-health authorities seem pretty well convinced that dietary polyphenols are an important part of good health, due to their supposed antioxidant properties. In the past, I've been critical of the hypothesis. There are several reasons for it:

1. Polyphenols are often, but not always, defensive compounds that interfere with digestive processes, which is why they often taste bitter and/or astringent. Plant-eating animals including humans have evolved defensive strategies against polyphenol-rich foods, such as polyphenol-binding proteins in saliva (1).
2. Ingested polyphenols are poorly absorbed (2). The concentration in blood is low, and the concentration inside cells is probably considerably lower*. In contrast, essential antioxidant nutrients such as vitamins E and C are efficiently absorbed rather than excluded from the circulation.
   
3. Polyphenols that manage to cross the gut barrier are rapidly degraded by the liver, just like a variety of other foreign molecules, again suggesting that the body doesn't want them hanging around (2).
4. The most visible hypothesis of how polyphenols influence health is the idea that they are antioxidants, protecting against the ravages of reactive oxygen species. While many polyphenols are effective antioxidants at high concentrations in a test tube, I don't find it very plausible that the low and transient blood concentration of polyphenols achieved by eating polyphenol-rich foods makes a meaningful contribution to that person's overall antioxidant status, when compared to the relatively high concentrations of other antioxidants in blood (uric acid; vitamins C, E; ubiquinone) and particularly inside cells (SOD1/2, catalase, glutathione reductase, thioredoxin reductase, paraoxonase 1, etc.).
   
5. There are a number of studies showing that the antioxidant capacity of the blood increases after eating polyphenol-rich foods. These are often confounded by the fact that fructose (in fruit and some vegetables) and caffeine (in tea and coffee) can increase the blood level of uric acid, the blood's main water-soluble antioxidant. Drinking sugar water has the same effect (2).
6. Rodent studies showing that polyphenols improve health typically use massive doses that exceed what a person could consume eating food, and do not account for the possibility that the rodents may have been calorie restricted because their food tastes horrible.
   

The main point is that the body does not seem to "want" polyphenols in the circulation at any appreciable level, and therefore it gets rid of them pronto. Why? I think it's because the diversity and chemical structure of polyphenols makes them potentially bioactive-- they have a high probability of altering signaling pathways and enzyme activity, in the same manner as pharmaceutical drugs. It would not be a very smart evolutionary strategy to let plants (that often don't want you eating them) take the reins on your enzyme activity and signaling pathways. Also, at high enough concentrations polyphenols can be pro-oxidants, promoting excess production of free radicals, although the biological relevance of that may be questionable due to the concentrations required.

A Reappraisal

After reading more about polyphenols, and coming to understand that the prevailing hypothesis of why they work makes no sense, I decided that the whole thing is probably bunk: at best, specific polyphenols are protective in rodents at unnaturally high doses due to some drug-like effect. But-- I kept my finger on the pulse of the field just in case, and I began to notice that more sophisticated studies were emerging almost weekly that seemed to confirm that realistic amounts of certain polyphenol-rich foods (not just massive quantities of polyphenol extract) have protective effects against a variety of health problems. There are many such studies, and I won't attempt to review them comprehensively, but here are a few I've come across:

• Dr. David Grassi and colleagues showed that polyphenol-rich chocolate lowers blood pressure, improves insulin sensitivity and lowers LDL cholesterol in hypertensive and insulin resistant volunteers when compared with white chocolate (3). Although dark chocolate is also probably richer in magnesium, copper and other nutrients than white chocolate, the study is still intriguing.
• Dr. Christine Morand and colleagues showed that drinking orange juice every day lowers blood pressure and increases vascular reactivity in overweight volunteers, an effect that they were able to specifically attribute to the polyphenol hesperidin (4).
• Dr. F. Natella and colleagues showed that red wine prevents the increase in oxidized blood lipids (fats) that occurs after consuming a meal high in oxidized and potentially oxidizable fats (5).
• Several studies have shown that hibiscus tea lowers blood pressure in people with hypertension when consumed regularly (6, 7, 8). It also happens to be delicious.
  
• Dr. Arpita Basu and colleagues showed that blueberries lower blood pressure and oxidized LDL in men and women with metabolic syndrome (9).
• Animal studies have generally shown similar results. Dr. Xianli Wu and colleagues showed the blueberries potently inhibit atherosclerosis (hardening and thickening of the arteries that can lead to a heart attack) in a susceptible strain of mice (10). This effect was associated with a higher expression level of antioxidant enzymes in the vessel walls and other tissues.

Wait a minute... let's rewind. Eating blueberries caused mice to increase the expression level of their own antioxidant enzymes?? Why would that happen if blueberry polyphenols were themselves having a direct antioxidant effect? One would expect the opposite reaction if they were. What's going on here?

In the face of this accumulating evidence, I've had to reconsider my position on polyphenols. In the process, and through conversations with knowledgeable researchers in the polyphenol field, I encountered a different hypothesis that puts the puzzle pieces together nicely.


* Serum levels briefly enter the mid nM to low uM range, depending on the food (2). Compare that with the main serum antioxidants: ~200 uM for uric acid, ~100 uM for vitamin C, ~30 uM for vitamin E.

Nitrate: a Protective Factor in Leafy Greens

Cancer Link and Food Sources

Nitrate (NO3) is a molecule that has received a lot of bad press over the years. It was initially thought to promote digestive cancers, in part due to its ability to form carcinogens in the digestive tract. As it's used as a preservative in processed meats, and there is a link between processed meats and gastric cancer (1), nitrate was viewed with suspicion and a number of countries imposed strict limits on its use as a food additive.

But what if I told you that by far the greatest source of nitrate in the modern diet isn't processed meat-- but vegetables, particularly leafy greens (2)? And that the evidence specifically linking nitrate consumption to gastric cancer has largely failed to materialize? For example, one study found no difference in the incidence of gastric cancer between nitrate fertilizer plant workers and the general population (3). Most other studies in animals and humans have not supported the hypothesis that nitrate itself is carcinogenic (4, 5, 6). This, combined with recent findings on nitrate biology, has the experts singing a different tune in the last few years.

A New Example of Human Symbiosis

In 2003, Dr. K. Cosby and colleagues showed that nitrite (NO2; not the same as nitrate) dilates blood vessels in humans when infused into the blood (7). Investigators subsequently uncovered an amazing new example of human-bacteria symbiosis: dietary nitrate (NO3) is absorbed from the gut into the bloodstream and picked up by the salivary glands. It's then secreted into saliva, where oral bacteria use it as an energy source, converting it to nitrite (NO2). After swallowing, the nitrite is reabsorbed into the bloodstream (8). Humans and oral bacteria may have co-evolved to take advantage of this process. Antibacterial mouthwash prevents it.

Nitrate Protects the Cardiovascular System

In 2008, Dr. Andrew J. Webb and colleagues showed that nitrate in the form of 1/2 liter of beet juice (equivalent in volume to about 1.5 soda cans) substantially lowers blood pressure in healthy volunteers for over 24 hours. It also preserved blood vessel performance after brief oxygen deprivation, and reduced the tendency of the blood to clot (9). These are all changes that one would expect to protect against cardiovascular disease. Another group showed that in monkeys, the ability of nitrite to lower blood pressure did not diminish after two weeks, showing that the animals did not develop a tolerance to it on this timescale (10).

Subsequent studies showed that dietary nitrite reduces blood vessel dysfunction and inflammation (CRP) in cholesterol-fed mice (11). Low doses of nitrite also dramatically reduce tissue death in the hearts of mice exposed to conditions mimicking a heart attack, as well as protecting other tissues against oxygen deprivation damage (12). The doses used in this study were the equivalent of a human eating a large serving (100 g; roughly 1/4 lb) of lettuce or spinach.

Mechanism

Nitrite is thought to protect the cardiovascular system by serving as a precursor for nitric oxide (NO), one of the most potent anti-inflammatory and blood vessel-dilating compounds in the body (13). A decrease in blood vessel nitric oxide is probably one of the mechanisms of diet-induced atherosclerosis and increased clotting tendency, and it is likely an early consequence of eating a poor diet (14).

The Long View

Leafy greens were one of the "protective foods" emphasized by the nutrition giant Sir Edward Mellanby (15), along with eggs and high-quality full-fat dairy. There are many reasons to believe greens are an excellent contribution to the human diet, and what researchers have recently learned about nitrate biology certainly reinforces that notion. Leafy greens may be particularly useful for the prevention and reversal of cardiovascular disease, but are likely to have positive effects on other organ systems both in health and disease. It's ironic that a molecule suspected to be the harmful factor in processed meats is turning out to be one of the major protective factors in vegetables.

Masai and Atherosclerosis

I've been digging deeper into the health of the Masai lately. A commenter on Chris's blog pointed me to a 1972 paper showing that the Masai have atherosclerosis, or hardening of the arteries. This interested me so I got my hands on the full text, along with a few others from the same time period. What I found is nothing short of fascinating.

First, some background. Traditional Masai in Kenya and Tanzania are pastoralists, subsisting on fermented cow's milk, meat and blood, as well as traded food in modern times. They rarely eat fresh vegetables. Contrary to popular belief, they are a genetically diverse population, due to the custom of abducting women from neighboring tribes. Many of these tribes are agriculturalists. From Mann et al: "The genetic argument is worthless". This will be important to keep in mind as we interpret the data.

At approximately 14 years old, Masai men are inducted into the warrior class, and are called Muran. For the next 15-20 years, tradition dictates that they eat a diet composed exclusively of cow's milk, meat and blood. Milk is the primary food. Masai cows are not like wimpy American cows, however. Their milk contains almost twice the fat of American cows, more protein, more cholesterol and less lactose. Thus, Muran eat an estimated 3,000 calories per day, 2/3 of which comes from fat. Here is the reference for all this. Milk fat is about 50% saturated. That means the Muran gets 33% of his calories from saturated fat. This population eats more saturated fat than any other I'm aware of.

How's their cholesterol? Remarkably low. Their total serum cholesterol is about half the average American's. I haven't found any studies that broke it down further than total cholesterol. Their blood pressure is also low, and hypertension is rare. Overweight is practically nonexistent. Their electrocardiogram readings show no signs of heart disease. They have exceptionally good endurance, but their grip strength is significantly weaker than Americans of African descent. Two groups undertook autopsies of male Masai to look for artery disease.

The first study, published in 1970, examined 10 males, 7 of which were over 40 years old. They found very little evidence of atherosclerosis, even in individuals over 60. The second study, which is often used as evidence against a high-fat diet, was much more thorough and far more interesting. Mann et al. autopsied 50 Masai men, aged 10 to 65. The single most represented age group was 50-59 years old, at 13 individuals. They found no evidence of myocardial infarction (heart attack) in any of the 50 hearts. What they did find, however, was coronary artery disease. Here's a figure showing the prevalence of "aortic fibrosis", a type of atherosclerotic lesion:


It looks almost binary, doesn't it? What could be causing the dramatic jump in atherosclerosis at age 40? Here's another figure, of total cholesterol (top) and "sudanophilia" (fatty streaks in the arteries, bottom). Note that the Muran period is superimposed (top).


There's clearly a pattern here. Either the Masai men are eating nothing but milk, meat and blood and they're nearly free from atherosclerosis, or they're eating however they please and they have as much atherosclerosis as the average American. There doesn't seem to be much in between.

Here's a quote from the paper that sums it up well:

We believe... that the Muran escapes some noxious dietary agent for a time. Obviously, this is neither animal fat nor cholesterol. The old and the young Masai do have access to such processed staples as flour, sugar, confections and shortenings through the Indian dukas scattered about Masailand. These foods could carry the hypothetical agent."

I know this blog is starting to sound like a broken record, but I'll say it again: you can eat a wide variety of foods and be healthy, except industrial grain products (particularly wheat), sugar, industrial vegetable oil and other processed food. The Masai are just one more example of a group that's healthy when eating a traditional diet.

What to do if Your Study Contradicts Conventional Wisdom

I just read a recent paper from the British Journal of Sports Medicine, "Daily Energy Expenditure and Cardiovascular Disease Risk in Masai, Ruran and Urban Bantu Tanzanians". The study caught my eye because I think we have a lot to learn from healthy traditionally-living populations worldwide.

The Masai have a very unique diet consisting almost exclusively of whole cow's milk, cow's blood and meat. As you might imagine, they eat a lot of fat, a lot of saturated fat and a modest amount of carbohydrate (from lactose). They also have low total cholesterol, low blood pressure, and virtually no overweight. They have been a thorn in the side of the lipid hypothesis for a long time.

The Bantu are an agricultural population that traditionally eat a diet low in fat and high in carbohydrate. Their staples are root vegetables, corn, beans, fish and wild game. The paper also describes a group of urban Bantu, which eats a diet intermediate in fat and carbohydrate. Incidentally, the investigators describe it as a "high-fat diet", despite the fact that the percentage fat is about the same as what Americans and Europeans eat, shamelessly exposing their bias.

The investigators recorded the three groups' diets, activity levels, physical characteristics and various markers of cardiovascular disease risk. Here's what they found: only 3% of Masai were obese, compared to 12% of rural Bantu and 34% of urban Bantu (they'd fit right in here!). The Masai, despite smoking like chimneys, had generally lower CVD risk factors than the other two populations, with the urban Bantu being significantly worse off than the rural Bantu.

Overall, the Masai came out looking really good, with the rural Bantu not too far behind. The urban Bantu look almost as bad as Americans. How do we make sense of these two conflicting facts? 1) The urban Bantu eat an amount of fat and saturated fat that's right in the middle of what the Masai and the rural Bantu eat, yet they seem the most likely to keel over spontaneously. 2) Saturated fat KILLS!! Answer: keep digging until you find something else to blame your results on.

They certainly did find something, and it's the reason the study was published in the British Journal of Sports Medicine rather than the American Journal of Clinical Nutrition. The Masai exercise more than either of the other two groups. I don't have too much trouble believing that. However, the authors used a dirty trick to augment their result: they normalized calorie expenditure to body weight. They present their data as kcal/kg/day. In other words, the fatter you are, the lower your apparent energy expenditure! It makes no sense to me. But it does inflate the apparent exercise of the Masai, simply because of the fact that they're thinner than the other two groups.

Due to this unscrupulous number massaging, here's what they got (data re-plotted by me):


I'm going to try to un-massage the data. Here's what it looks like when I factor bodyweight out of the equation. Calories expended (above resting metabolic rate) is on the Y-axis. The bars look a bit closer together...



Here's what it looks like when you add back resting metabolic rate. I assumed 1500 kcal/day. This graph is an approximation of their total energy expenditure per day:



Hmm, the differences keep getting smaller, don't they? I'm not challenging the fact that the Masai exercise more than the other two groups, but I do have a problem with this kind of manipulation of the data in misleading ways.

Their conclusion is that exercise is protecting the Masai from the deadly saturated fats in their diet. A more parsimonious explanation is that saturated fat per se doesn't cause heart disease. It's also more consistent with other healthy cultures that ate high-fat diets like the Inuit, certain Australian aboriginal groups, and some American Indian groups. It's also consistent with the avalanche of recent trials of low-carbohydrate diets, in which people consistently see improvements in weight, blood pressure, and CVD markers, among other things. Not that I have much faith in blood lipid markers of CVD.

My conclusion, from this study and others, is that macronutrients don't determine how healthy a diet is. The specific foods that compose the diet do. The rural Masai are healthy on a high-fat diet, the rural Bantu are fairly healthy on a low-fat, high carbohydrate diet. Only the urban Bantu show a pattern really consistent with the "disease of civilization", despite a daily energy expenditure very similar to the rural Bantu. They're unhealthy because they eat too much processed food: processed vegetable oil, processed grain products, refined sugar.

Thanks to kevinzim for the CC photo

Scientist Discovers that Only Pills can Control Hypertension

I went to a presentation today by a prominent hypertension researcher. His talk began with a slide that had two pictures side-by-side: one of the late fitness advocate Jim Fixx, and the other of Winston Churchill. Fixx was a marathon runner, while Churchill was inactive, overweight and had a famous appetite. Fixx died of a sudden heart attack at 52, while Churchill lived to 90. The presenter went on to state that this is an example of how genes control CVD risk, implying that despite Fixx's exemplary lifestyle, his genes had condemned him to an early death.

I wanted to jump up and yell "I think you're leaving out the alternate hypothesis: running marathons and stuffing yourself with grains isn't healthy!" But instead I suffered quietly through what ended up being an inane yet informative presentation.

His lab looks for gene variations that affect blood pressure (BP). There's a huge amount of money and research going into this. His lab and others have come up with two classes of mutations:

• Common allele variants that have an insignificant but measurable effect on blood pressure.
• Rare genetic mutations that have a significant effect on BP. The most common affects 1 in 2,000 people in the US.

Despite truckloads of funding and research, they have yet to uncover any gene or combination of genes that accounts for even a fraction of hypertension in Americans. So what's the next step? Keep looking for genes.

I suspect they will never find anything interesting. The reason? Hypertension is tightly linked to lifestyle. It's a quintessential aspect of the "disease of civilization". It's highly responsive to carbohydrate restriction, as a number of clinical trials have shown. Remember the Kuna? They don't get hypertension when they live a non-industrial, grain-free lifestyle (despite eating more salt than the average American), but as soon as they move to the city their hearts explode. It's been demonstrated in a number of other similar cases as well. Genetics are clearly not responsible.

Don't get me wrong, I do think genetics can modify a person's response to a poor lifestyle. But when the lifestyle is healthy, the vast majority of these differences fade away. I have a more thorough discussion of this point here.

If you give just the right dose of poison to a group of animals, 50% will die and 50% will survive (called the EC50 dose). You might then conclude that genetics had determined who lived and died. You wouldn't be wrong, but you'd be missing the point that what killed them was the poison.

The thing that really bothers me about this thinking is it's disempowering. The presenter suggested that the reason for the difference between Fixx and Churchill was their genes. If genes have us in such a tight grip, why bother trying to live well? The only logical solution is to pop hypertension pills and eat cake all day.

My guess is that if they had lived a more natural lifestyle, Fixx would have made it to 90 and Churchill would have been fit and lean.