Green Tea Dose-Dependently Sheds 7%-20% Body Weight in Mouse Model, But Higher Doses Turn Out to Be Pro- not Anti-Inflammatory In Recent Tufts Study

Image 1: A cup of freshly brewed
green tea (photo by Wikimol)

Green tea is healthy, it gets you going, will make you lose fat and calm down inflammation... wait! Is it possible that all that was just another marketing scam? Scientists from the Tufts University in Boston (Pae. 2011) have now found that at least in case of the capped green tea extracts, health (over-)conscious GNC-consumers are popping like candy, these days, the latter, i.e. the anti-inflammatory effect is absent.

And, it even gets worse, higher doses of a standardized EGCG extract (TEAVIGO; 95% EGCG content) even increased inflammation (measured via proinflammatory cytokines in the blood of the animals) in a group of 6-9 months old pathogen-free male mice who had been fed a diet enriched with 1% of the extract for 6 weeks:

Contrary to the assumption that EGCG would reduce inflammatory response, mice fed 0.15% and 0.3% EGCG diet exhibited no change while those fed 1% EGCG diet produced more proinflammatory cytokines tumor necrosis factor-α, interleukin (IL)-6, and IL-1β and lipid inflammatory mediator prostaglandin E2 in their splenocytes and macrophages (MΦ) and less IL-4 in splenocytes.

In that, it is quite interesting that the cytotoxicity of the green tea extract didn't hinder it from unfolding its unquestionably more marketable potential to reduce body weight.

Figure 1: Body weight of ECGC treated in the course of the 6 week treatment period; expressed relative to the weight of the unsupplemented control group (data calculated based on Pae. 2011)

As the data in figure 1 indicates, all three levels of EGCG enrichment produced statistically significant and surprisingly instantaneous reductions in body weight. The mean weight difference to the control group were -7%, -12%, -20% for the 0.15%, 0.3% and 1% EGCG groups, respectively. Based on previous research on the weight loss effects of EGCG extracts, Pea et al. hypothesize that the underlying mechanisms "involve decreased energy/lipid absorption and lipogenesis",  "increased fat oxidation" and improved glucose tolerance. Since the authors of respective studies were interested mainly in the weight loss effect, they failed to measure inflammatory status, as well, which is why Peat et al. are unable to tell, whether these beneficial metabolic effects are associated or even stem from EGCG's effect on inflammation status.

To evaluate the potentially detrimental effects of the wide-spread use of commercially available green tea extracts, the authors conduct the following dose-equivalent calculation:

Based on the average mice's consumption of 3 g/days, diets containing the medium dose (0.3%), or 3 g EGCG /kg diet, provided mice with a daily intake of 9 mg EGCG, or 300 mg/(kg BW/d) for a mouse of 30 g (average BW in the current study). When this dose is converted from mice consuming 12 kJ/day to humans consuming 2000 kJ/day by using isocaloric calculation, it is roughly equivalent to 22 mg/(kg BW/day) in humans, or 1540 mg EGCG per day consumed by a 70-kg person. Likewise, the low dose (0.15%) and high dose (1%) of EGCG for mice in this study are equivalent to human daily consumption of 770 and 5040 mg, respectively.

In view of the fact, a cup of tea contains only 150−180 mg EGCG, it is unlikely that habitual green tea (~3-5 cup = 450-900mg EGCG) consumption will increase daily intake to the "critical" level of  commercially available EGCG - it could, on the other hand, suffice to induce beneficial effects on body weight, glucose management and fatty acid metabolism.

Did you know that in many studies concentrations of 50µmol/L, i.e. roughly 230,000mg of epigallocatechin-3-gallate per liter, were used to elicit the often cited health benefits of the green tea phytochemical? In animal models, however, even a dose of 2,000mg/kg body weight incresed plasma concentrations to levels slightly below 1/5th (9µmol/L) of what has been studied in the petri dish.

In case of the consumption of commercially available EGCG caps, which contain anywhere from 150-900 mg EGCG/tablet (depending on capsule size and purity of the extract), the more-is-more mentality of our society along with the desire to lose weight as quickly as possible could however lead to epigallocatechin-3-gallate intakes at levels equivalent to those that were achieved in the study and would thus leave consumers slim, but heavily inflamed. So, if you want to avoid that, you better stick to the suggested serving size (in case of the NOW Foods EGCG product, for example, this practice would deliver ~600mg in three divided doses per 24h), instead of going through your cost-effective 250g bottle of bulk 90% EGCG extract within a single month :-)

Ask Dr. Andro: What Are Human Equivalent Doses (HED) and How Do I Calculate Them?

Image 1: Rat & guinea pig. Both very
popular animal models. Both about the
same size and weight, but with very
different reactions to identical treatments.

I won't waste too many words on this being a new series on the SuppVersity, I suppose the name "Ask Dr. Andro" is self-explanatory. It is based on questions posted in the comment section that are relevant for the majority of the visitors and require and deserve a more sophisticated response. So, if you want your question answered, learn a lesson from "Learner" who posted the following question in the comment section of "Spicing Up Fat Loss: Structural Similarity to Melanocortin-4 Agonists Powers Piperine's (Black Pepper Extract) Fat Loss and Lipid Lowering Effects".

Question from "Learner": Why is the mg/kg dosing ratio different for humans than rats, and does that vary by the agent?

Answer from Dr. Andro: Most of what we know about the effects of (new) drugs, supplements, vitamins, minerals, diets, macronutrient ratios and all the other stuff you read about here at the SuppVersity are derived from animal studies. The reasons thereof are manifold:

1. Animal models are "save" - you do not risk a human life, if you test your new drug on a rat, so this is "safe" for humans
2. Animal models are cheap and readily available - just order a bunch of lab mice, a few cages and the respective chow and get going; even in the days of slavery that would have been way more difficult with human beings
3. Animal models (at least some) are easily genetically modifiable - turn off a gene and see what it does
4. Animal models are time-efficient - if you want to know how life-long supplementation with substance A affects a critter, you better not use a tortoise or your experiment will literally "survive" you
5. Animal models have an endless list of other advantages, I do not want to list here, because they are not really relevant for the question at hand...

Notwithstanding all these benefits, as a faithful visitor of the SuppVersity you will be familiar with the problems that arise in interpreting data from studies conducted on one species in view of expected results of similar experiments conducted with another species. Even rats and mice can react very differently to a given substance and despite their similar weight and seize guinea pigs and rat models can by no means be used interchangeably.

But let's get back to the "human equivalent" - obviously, two of the most apparent differences between human beings and rats, are body size and body weight. As you probably know, most studies provide information about dosages relative to body weight, so that, for the reader, the difference in body weight would not matter, anyways. Unfortunately a direct translation of the effects observed in a rodent model to a human scenario, based solely on body weight turns out to be very misleading.

Example 1 (this is how it is not done): A very prominent example, where the futile effort of calculating human equivalent doses simply by multiplying the given dosages with the weight of an average human being, lead to major (public) confusion, were the studies on the beneficial effects of the red wine polyphenol resveratrol by Joseph A. Baur et al. (Baur. 2006) and Marie Lagouge et al. (Lagouge. 2006) that were interpreted by the "laypress" as meaningless, because the purported "equivalent" dosages (as mentioned before, calculated solely by multiplying the dose used in the study by the weight of an average human being, cf. figure 1) would amount to hundreds, if not thousands of liters of red wine per day.

Figure 1: Example of a simplistic (and inappropriate) calculation of the human dose equivalent.

This procedure, apparently fails to take into account that oxygen utilization and caloric expenditure are much greater in rodents than in human beings, or in other words: rodents burn through food and metabolize drugs, supplements and whatever you feed them much faster than humans. In the very extreme this may mean that what is a very small dose kilogram per kilogram for a mouse, may well kill an adult human being within minutes.

Image 2: Max Rubner was
probably the first to collect
reliable data on the correlation
of surface area, caloric expen-
diture and oxygen consumption
in mammals (image from The
Eskind Biomedical Library
at the Vanderbilt University)

Thanks to the meticulous research of the German scientist Max Rubner (Rubner. 1883), we do yet know that, in most mammals, both oxygen utilization and caloric expenditure correlate quite well with the ratio of the surface area of the critter to its body weight. An average laboratory mouse, for example, has a very small body weight of ~20g and a comparably large surface area of roughly ~0.006m². An average, normal weight male human being (height 1.73m), on the other hand, weighs roughly ~70kg and has a relatively small surface are of ~1.9m². This means that a man weighs about 3500x as much as a mouse, yet his surface area is only 316x larger than that of the small rodent.

You may now ask yourselves how all that relates back to energy expenditure and metabolic rate - well, no matter how thick the fur of the mouse may be, there is no way for the little critter to escape the basic laws of thermodynamics, which state that the heat flow (for the mouse heat loss) across the surface of an object is proportional to its surface area. On a pound per pound base, the mouse will thus expend way more energy than the man just to keep its body from cooling down.

With that being said, it should be quite obvious that the ratio of body weight [in kg] to surface area [in m²], the so called K_m factor, can be used as a relative measure of the metabolic rate of an animal. Using the previously given data, we can now calculate K_m factors of K_m =3 and K_m=37 for mouse and man, respectively. Then, the ratio of the two, K_m(mouse)/K_m(man) = 0.081 = 8.1%, indicates that the human metabolism runs at 8% of the metabolic rate of a mouse, or in other words, mice metabolise whatever supplement or medication you feed or inject them at a 12 times higher rate than human beings. If you compare that to the respective respiratory ratios of 165 breaths per minute for mice and 12-20 breaths per minute for healthy adults, the aforementioned correlation between body weight and surface area on the one hand and metabolic rate and oxygen consumption on the other becomes quite obvious.
Now, we can save ourselves future troubles associated with the experimental determination of the aforementioned metabolic parameters for each and every species (and animals of the same species, yet with different body sizes / form, e.g. children vs. adults) if we just collect the respective data on body weights and surface areas, and tabulate the respective K_m values. Using these values we can easily calculate the equivalent doses for a given species by using the following equation (cf. Reagan-Shaw. 2008):

HED (mg/kg) = Animal dose (mg/kg) * (Animal K_m : Human K_m)

Where the term in (mathematically unnecessary brackets) would be a conversion factor given by the ratio of the K_m value of the species that was used in the respective study to the K_m value of the target species. For the majority of the studies you will find on the SuppVersity, the original study was done on mice (K_m = 3) or rats (K_m = 6) and the conversion factors which are probably of greatest interest for you, i.e. the ones for humans would be 3/37=0.081 for mice (cf. discussion above) and 6/37=0.162 for rats, respectively.

The people from the FDA, who always tend to think nobody could be half as smart as they are, obviously wanted to play safe and included these "conversion factors" along with the K_m values in their Guidance for Industry on "Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers" (FDA. 2005):

Table 1: Km values for various species according to FDA Guidance for Industry (FDA. 2005)

Example 2 (this is how it is done): With this new knowledge you can also calculate how much black pepper you would have to consume to achieve the effective dosage of piperine that was used in the study mentioned in the SuppVersity news last Thursday. With an average piperine content of 3 to 8 g/100g and about 0.2-0.3g/100g of the minor alkaloids piperyline and piperettine in black pepper (Schulz. 2004), a rat would have to consume somewhere between 500-1300mg of black pepper corns per day to achieve the 40mg/kg dose that effectively accelerated the metabolism of the rats in the study by Shat et al. (Shah. 2011). All you have to do now, is look up the K_m values of rats (K_m=6) and humans (K_m=37) and insert the respective values in the given equation, so that

HED(40mg/kg in rats) = 40mg/kg * (6 kg/m² : 37kg/m²) = 6.5mg

would yield the amount of piperine and

HED(1000mg/kg in rats) = 1000mg/kg * (6 kg/m² : 37kg/m²) = 162mg

the amount of pepper corns you would have to use to achieve the "same effect" in human beings.

A final word on the italicized "same" in what would otherwise be the last sentence of this article: You will probably never see identical effects across different species, regardless of how exact you try to calculate the equivalent doses. There are simply too many differences between mice and men and the way they metabolize different substances to be able to translate results from experiments with oftentimes fundamentally different agents from mice to men and vice versa using nothing but the ratio of body weight to surface area as your guide. Consequently, you should consider human equivalent doses as a means to interpreted whether or not animal data is practically relevant and what amounts of a given substance would roughly have to be used in follow up studies to evaluate whether or not the results can be reproduced in human beings.

Leptin & AMPK Activation in Muscle - Did Scientists Investigate the Wrong Species?

Actually, this is something I have been reminding people of for years: Human beigns are no mice and although some people may be rats, genetically they are still very different. Usually I put this argument forward if anyone presents me with a study done on rodents that "shows" how bad dietary protein, especially from meat, is for you - thin of it: when was the last time you saw a mouse eating a cow? This time the issue is a little more sophisticated, though...

Australian scientists (Laker. 2010) have recently investigated the effect of leptin on AMP-activated protein kinase (AMPK) in muscle of sheep and (I do not know if to their, but maybe to some of the proclaimed experts surprise) they found that...

[...] leptin infusion reduced (P<0.05) food intake and body weight and it also increased plasma adrenaline concentration at 6 h and 7 days, suggesting increased sympathetic nerve activity. Despite this, and in contrast to rodent studies, central leptin infusion did not increase skeletal muscle AMPKα Thr172phosphorylation or ACCβ Ser221 phosphorylation.

With sheep being genetically closer to human beings than mice, this is an interesting finding and could possible lead into new insights into why leptin, once proclaimed as possible panacea for the obesity epidemic, does work on rodents, but has little positive effect on obese human beings. Hitherto, the most accepted theory is "leptin resistance", which, similar to insulin resistance, is supposed to prevent leptin from doing its magic. This study, however, suggests that there might be genetic issues involved, as well. As always, I will keep you posted...