Inflammation Is a True Fat Burner: BSO-Induced Glutathione Depletion Wards off Fat Gains on Hypercaloric Diet

Image 1: This little bugger obviously has too little inflammation going on ;-)

Are you "on fire"? Inflammation has been implicated as the root cause of almost all modern disease: obesity, diabetes, heart disease, cancer, you name it. Soothing the flames via natural and supplemental anti-oxidants has thusly been proposed and marketed as a solution for many of the aforementioned health problems.

Yet, despite tons of vitamins, anti-oxidants and all the other "healthy" stuff we are taking and consuming on a daily basis, the number of morbidly obese people, diabetics and heart attack patients appears to be ever-increasing... how can that be?

A possible answer to that question comes from scientists from the Saha Cardiovascular Research Center at the University of Kentucky College of Medicine in Lexington, Kentucky, US (Findeisen. 2011) - we simply got everything wrong! The observation that insulin resistance and beta-cell dysfunction usually occur in the presence of large amounts so-called reactive oxygen specimen (ROS) lead scientists to propose that there was a causative relationship between these two events, of which the data only shows that they are corollary.

Image 2: Whenever there is a fire, the firefighters are not far away, but does this correlation indicate that all firefighters are firebugs? (img texarkanagazette.com)

Despite the fact that the distinction between correlation and causation should be obvious, correlations have a long history of being mistaken as causative factors in the history of science. The corollary elevation of total cholesterol in heart disease patients, for example, is the reason that millions of well-educated people world-wide still believe that cholesterol would cause heart disease - an erroneous conclusion for which my friend, Carl Lenore, has coined a very fitting analogy (actually the analogy spans all those "corollary causation"): When there is a fire in down-town New York, it won't take long until the place is packed with firefighters, nevertheless, no sane observer would get the idea that the corollary appearance of firefighters on the scene would be the reason for the fire.

Here, at the SuppVersity, you have already learned that a group of researchers from Germany has invested a lot of work into research on the beneficial effects of inflammation (Ristow. 2010). Now, with the data from Hannes M. Findeisen (who unquestionably is a German or has German ancestors, as well ;-) et al., evidence begins to accumulate that the role of reactive oxygen specimen in glucose homeostasis could in fact be a beneficial and not a detrimental one. After all, Findeisen and his colleagues were able to show that the pharmacological depletion of glutathion, our natural broadband fire-extinguisher, made mice resistant to diet-induced obesity, increased energy expenditure and enhanced insulin sensitivity.

If you have listened to all the installments of the Amino Acids for Super Humans Series on Carl Lenore's Super Human Radio, you will already have heard me mention that a methionine/cysteine-free diet has been shown years ago to have profound fat-burning, or I should say, weight-reducing effects on mice - no wonder, with methionine and cysteine being essential substrates for mammalian gluthation production, a lack of these dietary sulfur-amino acids induced a similar glutathion depletion as the addition of 30mmol/l BSO to the drinking water of the mice in the Findeisen study (for more on the glutathion depleting effects of BSO, cf. Skapek. 1998; Mira. 2002; Cattan. 2008)

Even before the works of Ristow et al. and now Findeisen et al., it has been well-established that reactive oxygen specimen, the purported villains of the 21st century, enhance cellular signaling (Veal. 2007). About a year ago, Chang and Chang  reported that H2O2, in particular, is a potent activator of protein signaling pathways, including insulin signaling and can even mimic insulin's effects by the inhibition of oxidation-sensitive protein tyrosinases (Chang. 2010). With glutathion being the primary H2O2 scavenger in mammalian tissue, it is thus not surprising that the BSO treated and thusly glutathion depleted mice in the Findeisen study displayed a more favorable response to a glucose tolerance test after 6 weeks of treatment with BSO and a 45%(high)-fat diet (cf. figure 1).

Figure 1: Glucose levels in mg/dl after oral glucose tolerance test in mice after 6 weeks on a high-fat diet (47% fat) with or without 30mmol/L BSO in their drinking water (data adapted from Findeisen. 2011)

These results are surprising, also because the daily food and water intake of the mice was identical. The latter cannot be said of their calorie-expenditure, daily activity level (cf. figure 2) and the activity of the "fat burning" uncoupling protein UCP2 (+100%), the elevation of which increases thermogenesis and energy expenditure.

Figure 2: Relative changes in energy expenditure and daily activity due to BSO induced glutathion depletion in mice on a high fat diet compared to non-treated control (data calculated based on Findeisen. 2011)

Now, most importantly for you, as a physical culturist, may be that glutathione depleted mice did not simply fail to thrive or shrivel away - they were, as Findeisen points out...

completely protected from diet-induced obesity, despite similar food intake and water consumption. Analysis of body composition in mice fed a HFD diet confirmed significantly decreased fat mass in BSO-treated mice without significant differences in lean body mass, indicating that the difference in body weight was due to reduced fat mass in BSO-treated mice.

If you don't believe the words, I suggest you take a look at the data in figure 3 - while the control mice had a body fat percentage of whopping 32% the mice on BSO with their ~16% body fat were well within the normal range for lab-mice.

Figure 3: Fat and lean mass (in g) of mice from the control group and the glutathion-depleted group after 6 weeks on a hypercaloric high fat diet (data adapted from Findeisen. 2011)

Even the researchers appeared to be surprised by the profound effects glutathion depletion had on the rodent's ability to accumulate body fat. As far as the underlying reasons are concerned, they speculate that it was ...

[...] possible  that  the  observed  increase in the expression of UCP-2 and UCP-3 in BSO-treated mice induced  mitochondrial  uncoupling [...] Alternatively, the enhanced energy expenditure in BSO-treated mice  might  be  the  result  of  increased  locomotor  activity.  In skeletal muscle, ROS are necessary for optimal contractile function, force production, and exercise-induced adaptations. Furthermore, particularly H2O2 is increasingly recognized as
a potent neuromodulator. It is therefore conceivable, that glutathione depletion may lead to activity-stimulating changes in the redox environment of muscle or brain.

Now, it is however questionable in how far any of these three phenomena would occur in human beings, as well. While the lack of large amounts of UCP-sensitive brown adipose tissue would decrease the UCP induced thermogenic response to glutathione depletion, locomotor activity is something that appears to be completely blocked in the modern couch potato, anyways. It would thus warrant further research (and studies into the general safety of this approach) before it would appear warranted that you take a spoon of BSO with every meal to counter the negative effects of your last binge ;-)

Not All Vitamin C is Created Equal: AA-2βG, a Powerful Vitamin C Analogue From Goji Berries Outperforms Its Cousin L-Ascorbic Acid and Teaches Scientists "Nature Still Knows Best!"

Vitamin C, also known as L-ascorbic acid probably is the best known of all anti-oxidants; and the marketing  departments of the food companies know that and how to make use of its popularity with slogans like "Extra rich in vitamin C", "Extra Vitamin C", etc. Back in the days, when food was still exclusively nourishing and nobody expected it to heal the ailments it, or other food was causing, ascorbic acid was mostly added to products to extend their shelf-life (this is still common practice, btw.). Today, however, the highly processed foodstuff the unhealthy majority of the fast food society, we have become, is consuming on a daily basis contains vitamin C to... well, I guess to be more marketable. After all, scientific evidence for the purported beneficial effects of vitamin C in isolation, i.e. outside of the natural nutrient mix of real food (vegetables, fruits, meat, eggs, etc.) is scarce and a recent study (Zhang. 2011) from the College of Life Science at the Ningxia University in Yinchuan, Ningxia, China, suggest that structural and compositional differences between vitamin C as we know it, i.e. l-ascorbic acid, and the naturally occurring mix of ascorbic acids and its structural analogues may be the actual reason for the lack of effect all the added ascorbic acid in our foodstuff has on the health of its consumers.

Image 1: Dried Goji or Wulfberries, a
natural source of powerful VitaminS(!) C.
1 gram of the dried fruit contains about
5mg of the power-antioxidant AA-2βG
(data from Toyoda-Ono. 2004)

The paper (Zhang. 2011), which appeared in the May issue of the Archives of Pharmacological Research, reports the results of in-vitro and in-vivo analyses of the anti-oxidant activities of AA-2βG, a natural vitamin C analogue from Goji berries (Lycium barbarum L.). From a molecular perspective, 2-O-β-D-Glucopyranosyl-L-ascorbic acid (AA-2βG) is nothing but plain ascorbic acid (AA) with an added D-glucose moiety and a β-glucoside linkage at the C2 position of the AA molecule. While the fact that this "extension" should (see below) reduce the count of hydrogen radicals or electrons the molecule can donate to scavenge NO2- molecules from two (AA) to one (AA-2βG), it adds to the stability of the molecule, which in its original form (ascorbic acid) cannot "is poorly stored in the body [which] makes it difficult to sustain high concentrations of AA within the body for therapeutic interventions". So, other than science, nature, in her infinite wisdom ;-), obviously knew about the storage problem with vitamin C all along and hid the solution in a fruit that, despite having being used in traditional Chinese medicine for hundreds of years, appeared on the screen of western medical science (and in a huge amount of commercial supplements) only very recently: Goji berries, lat. Lycium barbarum L., the dry fruit of L. barbarum (cf. image 1).

For the alpha- variety, AA-2αG (also known as AA-2G), which has a D-glucose moiety and an α-glucoside linkage at the C2 position, studies similar to the one performed by Zhang et al. had already found that despite the reduced amount of donable hydrogen radicals or electrons (see above), compared to normal ascorbic acid, the AA-2G radical that formed in the process of a first DPPH  [2,2-diphenyl-1-picrylhydrazyl is a dark-colored crystalline powder composed of stable free-radical molecules used to test the in-vitro anti-oxidant capacities of various chemicals compounds] scavenging was "able to react with another DPPH molecule to form a covalent adduct", this covalent adduct from the second reaction was then capable of "slowly quenching a third DPPH radical molecule to generate an unidentified product", so that one molecule of this vitamin C analogue, despite ostensible structural inferiorities, scavenges one additional DPPH radical compared to plain L-ascorbic acid. And while the 2-O substituted AA derivative does lack the ability to to scavange O2−radicals in vitro,

[...] it was more efficient at scavenging H2O2 and OH- than AA (p < 0.01 and p < 0.05, respectively) (Figs. 2B, 3A). This implied that the AA-2βG and AA [ascorbic acid] antioxidant mechanisms differ such that the antioxidant activities of AA-2βG are efficient at scavenging H2O2 and OH- instead of directly scavenging O2−radicals.

With different mechanisms of action, however, the radical scavenging effects of L-ascorbic acid and its derivative(s) add up. This makes the naturally occuring mix of vitamins C an even more potent weapon in the nutritional, supplemental and pharmacological war against oxidative stress reduction and the prevention of lipid peroxidation and cell damage (e.g. erythrocyte hemolysis; cf. figure 1).

Figure 1: Percentage of hemolytic red blood cells in a solution with 500µM H2O2 at various concentrations of Ascorbic Acid (AA) and AA-2βG cells vs. control (0µM) without added anti-oxidants (data adapted from Zhang. 2011)

If you bear in mind that even in the absence of H2O2 roughly 9% of the erythrocytes are damaged by oxidation, the level (~13%) at which the protective effects of AA-2βG saturate (cf. figure 1) indicates that the vitamin C analogon blocks H2O2 induced hemolysis almost completely.

Figure 2: Liver-protective effect of different doses of AA-2βG(dose in mg/kg) in CCl4-induced mouse injury
model; ALT, AST on primary, MDA on secondary axes (data adapted from Zhang. 2011)

Similarly astonishing are the liver protective effects of AA-2βG, of which Zhang et al. found in experiments with CCl4 intoxicated rodents that, in contrast to untreated animals,

[...] serum ALT and AST levels in animals pretreated with AA-2βG were reduced significantly compared to those in the untreated group (Table III) [and] AA-2βG exhibited dose-dependent protection against liver injury, as serum ALT and AST activities in mice given a high AA-2βG dose (300 mg/kg) decreased dramatically to levels similar to those in untreated animals.

So, after all, this seems to be another instance, where nature knew best. Even with all the bioflavonoids, esters, minerals or whatever else supplement producers keep adding to plain L-ascorbic acid to justify the exorbitant prices of their products, the natural vitamin C mix in a bunch of fresh Goji or Wulfberries still outperforms the fanciest supplement.