Overtraining, Inflammation, Insufficient Repair: Scientists Shed Some More Light on the Counterproductive Triad of Ups & Downs in Testosterone, IL-6, IL-10, COX II & Co

Image 1: Part of Christian Bale's protocol to become the skinny Machinist was, you guessed it, overtraining - accompanied by undereating, the results (left) were profound.

The debate on the practical implications and even the existence of a certain physical condition that is generally referred to as "overtraining" is probably one of the most longstanding debates in the realms of physical culture. From a scientific point of view, it stands out of question that the same exercises that turn a stringbean into a heavily muscled berserk can also change him back into a stringbean, when the narrow yet tremendously productive margin between "just enough" and "already too much" is repeatedly exceeded. Scientists from the Department of Sports Medicine at the Shanghai University of Sport probed the underlying causes of this phenomenon and came up with a few interesting observations (Xiao. 2012).

Update: My friend Carl Lanore from Super Human Radio happened to read this post, yesterday and as chance would have it, had a show scheduled with Brooks Kubik on the issue of Overtraining and How it Pertains to General Health, he called me and we had a nice round-table discussion on air. So, in case you are interested, here is the podcast.

Overtraining is determined by an imbalance of pro- and anti-inflammatory factors

While many of  the results of their 11-week rodent study are not actually new (e.g. weight loss, drop in testosterone, etc. figure 1) and their significance for the average physical culturist, who won't be following an exclusively treadmill based exercise protocol are questionable, their findings pertaining to the cytokine response to overtraining could shed some light onto the underlying molecular underpinnings of both skeletal muscle atrophy and hypertrophy.

Figure 1: Study design (left) and relative changes in body weight, hemoglobin and testosterone compared to sedentary control in the overtrained rodents (data calculated based on Xiao. 2012)

It does not take much to identify the profound imbalance in pro- and anti-inflammatory cytokines the three weeks of excessive exercise brought about (cf. figure 2). In order to be able to understand how these changes affect your gains and eventually even make it impossible for your body to repair the damage you are doing on a daily basis, it is yet important to understand what the physiological role of the individual cytokines is.

Figure 2: Changes in inflammatory and anti-inflammatory factors (left) and change in gastrocnemius weight and ratio of body weight to gastrocnemius weight in the overtrained rat immediately after the protocol (OT) or after recovery (OTR; data calculated based on Xiao. 2012)

While most of you will probably be familiar with the "bad guys", IL-6, TGF-beta1, the physiological role of the "good guys", interleukin 10 (IL-10), cyclooxygenase II (COX II) or the more or less exotic urokinase type plasminogen activator (uPA) could be all Greek to you, even though they may be the ones which make the exercise induced gains we often take for granted possible:

• IL10 is the "calming" counterpart to IL-6 and co. it acts directly on monocytes and inhibits the synthesis of pro-inflammatory cytokines such as IFN-γ, IL-2, IL-3, TNFα & Co.
   
• COX in particular has been identified as a necessary factor for satellite cell activity (Hill. 2003) and muscle regeneration (Bondesen. 2004)
• The inhibition of COX activity by ingestion of nonsteroidal anti-inflammatory drugs (NSAIDs) has been shown to to suppress the increase in mixed muscle protein synthesis due to exercise (Trappe. 2002) 
• COX-2 null mutants also showed less macrophage invasion of injured muscle during regeneration (Bondesen. 2004), a factor about which you should have read in the Intermittent Thoughts on Building Muscle (Part II) that is a critical factor for the recruitment and "installation" of satellite cells into damaged or growing muscle tissue.
   
• Urokinase type plasminogen activator (uPA) promotes the migration of numerous cell types, including macrophages, activated peripheral blood monocytes, endothelial cells, smooth muscle cells, and myoblasts (Novak. 2004). 
• Several studies have shown that the accumulation of macrophage the damaged muscle was impaired in uPA-null mice (Lluis. 2001; Koh. 2005), with the result being impaired repair of the muscle tissue.
• uPA increase the so-called hepatocyte growth factor, a paracrine signalling molecule that tells progenitor cells to "get going" and is required for skeletal muscle regeneration (Sisson. 2009), as well as the proliferation, migration and fusion of satellite cells (Bonavoaud. 1998; Munoz-Canoves. 1997; Fibbi. 2001) . 

You could thus summarize the results as follows, the profound skeletal muscle damage cannot be repaired, because

1. the downregulation of IL-10 exasperates the inflammatory reaction to exercise induced muscle damage
2. the reduction of COX II inhibits or mitigates the protein synthetic response to exercise
3. low COX II and uPA levels counteract the necessary replacement of damaged, let alone the installment of new myonuclei from quiescent progenitor cells (satellite cells)

The results are profound and, as the body weight to gastrocnemius weight ratio in the OTR group suggests, had detrimental effects on the body composition of the rodents (cf. figure 2, right). So, unless your goal is to be skinny and you are willing to overtrain to stay that way for the rest of your life, you better stick to a reasonable training volume and allow your body the rest it needs to recover and grow.

Practical implications: Questionable

Yet while the study results should have made it pretty clear and yes, this means dragging yourself to the gym with delayed-onset-muscle soreness (DOMS) that requires "treatment" with NSAIDs is no longer an option, it is of little help to determine where exactly the initially mentioned margin begins and where it ends. I believe I have given you a couple of good starting points in the Step By Step Guide to Your Own Workout Regimen, but in the end, the number of factors which will influence both the position as well as the width of this margin are so numerous that is would be unrealistic to assume that any study, rodent or human, aerobic or anaerobic, one or twelve week, ... will ever provide you with the answer to the question I know you were just about to type into the comment area of this post: "Am I already training too much? Or would it be better if..."

Figure 3: Overview of selected biomarkers that have been investigated for their usefulness to identify over-reaching or overtraining (first published in "The Overmotivational Roots of Overtraining"; based on an overview in Purvis. 2010)

What could yet potentially come out of this data is a test kit, one which would probably be a more reliable indicator of overtraining than DOMS or the creatine kinase levels which have long been touted as a potential candidate to distinguish between load and overload (cf. figure 3 and "The Overmotivational Roots of Overtraining"). Whether we will see respective test kits being available for the average Joe or Jane, is yet as questionable as whether the latter won't still rather waste his / her money on the latest and greatest supplement scam than on an optimized training routine. After all, a good personal trainer should - without any fancy tool-kits - be able to prescribe a routine that may not be 100% optimal, but will at least keep you within the repeatedly mentioned margin of productive overload, today!

References:

• Bonavaud S, Charriere-Bertrand C, Rey C, Leibovitch M P, Pedersen N, Frisdal E, Planus E, Blasi F, Gherardi R, Barlovatz-Meimon G. Evidence of a non-conventional role for the urokinase tripartite complex (uPAR/uPA/PAI-1) in myogenic cell fusion. J Cell Sci 1997; 110 : 1083 – 1089
• Bondesen B A, Mills S T, Kegley K M, Pavlath G K. The COX-2 pathway is essential during early stages of skeletal muscle regeneration. Am J Physiol 2004; 287 : C475 – C483
• Fibbi G, Barletta E, Dini G, Del Rosso A, Pucci M, Cerletti M, Del Rosso M. Cell invasion is affected by differential expression of the urokinase plasminogen activator/urokinase plasminogen activator receptor system in muscle satellite cells from normal and dystrophic patients. Lab Invest 2001; 81 : 27 – 39
• Hill M, Goldspink G. Expression and splicing of the insulin-like growth factor gene in rodent muscle is associated with muscle satellite (stem) cell activation following local tissue damage. J Physiol 2003; 549:409 – 418  
• Koh T J, Bryer S C, Pucci A M, Sisson T H. Mice deficient in plasminogen activator inhibitor-1 have improved skeletal muscle regeneration. Am J Physiol 2005; 289 : C217 – C223 
• Lluis F, Roma J, Suelves M, Parra M, Aniorte G, Gallardo E, Illa I, Rod-riguez L, Hughes S M, Carmeliet P, Roig M, Munoz-Canoves P. Urokinase-dependent plasminogen activation is required for efficient skeletal muscle regeneration in vivo. Blood 2001; 97 : 1703 – 1711
• Munoz-Canoves P, Miralles F, Baiget M, Felez J. Inhibition of urokinase-type plasminogen activator (uPA) abrogates myogenesis in vitro. Thromb Haemost 1997; 77 : 526 – 534
• Novak M L, Bryer S C, Cheng M, Nguyen M H, Conley K L, Cunningham A K, Xue B, Sisson T H, You J S, Hornberger T A, Koh T J. Macrophage-specific expression of urokinase-type plasminogen activator promotes skeletal muscle regeneration. J Immunol 2011; 187 : 1448 – 1457 
• Sisson T H, Nguyen M H, Yu B, Novak M L, Simon R H, Koh T J. Urokinase-type plasminogen activator increases hepatocyte growth factor activity required for skeletal muscle regeneration. Blood 2009; 114 : 5052 – 5061
• Trappe T A, White F, Lambert C P, Cesar D, Hellerstein M, Evans W J. Effect of ibuprofen and acetaminophen on postexercise muscle protein syn-thesis. Am J Physiol 2002; 282 : E551 – E556
• Xiao W, Chen P, Dong J. Effects of Overtraining on Skeletal Muscle Growth and
  Gene Expression. Int J Sports Med. 2012 May 16. [Epub ahead of print]

Science Round-Up Seconds: Cytokine Modulating Power Cakes, Endurance & Resistance for Maximal Cardiovascular Benefit, Gluten Degrading Bacteria and Cancer Fighting Green Tea + Levitra Combo. Plus: Sirt-3 & Protein Blends

Not sure if the "Power Cakes" in the Kerasioti study looked like these, but it would sure have been appropriate, after all the poor study participants had to cycle for 3h (in total) - makes you wonder if they are WADA approved, doesn't it?

Today's installment of the SuppVersity Science Round-Up Seconds has some delicious stuff for you in stock. We will start out with an interleukin 6 & 10 modulating CRP reducing cake from Greece as an appetizer, or pre-workout, whatever you like better. When we have finished that one, the table or gym talk will center around debunking the myth that only aerobic training was good for your cardiovascular system. When even the last i**** has gotten that, it's about time to devour the delicious anti-carcinogenic and gluten destroying bacteria stew with green tea and Levitra(R), I've been cooking for ya ;-)

Before you go for the cake, however, I want to apologize that I was somewhat out of it on yesterday's show (click here to listen to the podcast). As a compensation today's Seconds have - as you can see - become pretty "nutritious" at least on a quantitative level ;-)

Looking for the perfect peri-workout meal? What about some cake?

(Kerasioti. 2013) -- It may not really sound like "high performance fuel", but in fact the protein cake a group of Greek scientists fed to their nine physically active and pretty well-conditioned male subjects (age, 28 ± 2 yr; height, 184 ± 3 cm; body fat, 11 ± 2%; body mass index, 23 ± 1 kg/m²) turned out to be much more than just performance fuel after the 2 h of continuous cycling on cycle ergometer at an intensity corresponding to 60-65% of the subjects established VO2max.

Figure 1: Interleukin 6, interleukin 10 and c-reactive protein (CRP) levels before and after the first 2h exercise bout with experimental (EXP) and placebo (PLA) cake; data expressed relative to baseline (Kerasioti. 2013).

As the data in figure 1 goes to show you, the cakes which contained a combination of 0.9g/kg carbohyrates and 0.26g/kg protein did in fact have surprisingly more beneficial effects on the rise and fall of the pro- and anti-inflammatory cytokines IL-6 and IL-10 (IL = interleukin) during the two counterbalanced trials, than the placebo, an identical cake with a different macro-nutrient ratio of 1.1g carbs and 0.1g protein per kg body weight.

Training twice a day? No problem with the power cake ;-)
 
Only four hours later the scientists shooed their subjects onto the bike again for another hour of medium intensity exercise and a 95% VO2max time-trial sprint at the end, "to determine if the cake administration affects performance" (Kerasioti. 2013). Sounds logical, right? What's confusing, though is that Kerasioti et al. don't even mention the performance effects they did or didn't observe in the second bout in the discussion of their results - so I suspect that (a) there were no differences or (b) Kerasioti et al. regard the cytokine response to the second trial as a "performance" marker and stick to that as their "effect on performance" (I don't know about you, bun in my humble opinion the latter would be an even greater flaw than leaving the non-significant information out).

Figure 2: IL-6, IL-10 and CRP response over the whole study period; expressed relative to baseline (Kerasioti. 2013)

If you add the cytokine expression in the steady state 2h trial from figure 1 to the data that was recorded after the completion of the 1h steady state + time trial cycling session four hours later and the 48h post levels of the inflammatory markers, the resulting graph exposes a very neat symmetry with an almost identical rise and fall in the expression of the short-lived cytokines and an eye-catching difference in the intermediate inflammatory response evidenced by the increase in CRP in the recovery phase of the placebo trial (just as a reminder: C-reactive protein aka CRP is the same stuff you will also see chronically elevated in diabesity and all sorts of inflammatory diseases).

A propos protein: We also talked about the benefits of protein blends with fast and slow proteins.Here is the SuppVersity article about the human study on the casein + whey combination I mentioned.

Bottom line: Compared to carbohydrates alone the combination of carbs + whey does not only offer a muscle building, but also an inflammatory edge, especially endurance athletes and anyone with a high training density will benefit from -- just like the 3g glutamine (1.5g pre, 1.5g post workout), Carl and I talked about during yesterday's installment of the SuppVersity Science Round-Up, by the way, with the only difference being that the effects of whey protein are probably mediated by it's glutathione (the master oxidant) repleting glutamylcysteine groups (Bounous. 1991), beta-lactoglobulin, lactoferrin and other anti-inflammatory compounds in whey proteins (Peng. 2009).

Cardio and weights: Your cardiovascular system loves both

(Spence. 2013) -- There is this longstanding and die hard myth that only aerobic training would be good for the heart. With the publication of a recent paper by a group of scientists from the University of Western Australia and the Liverpool John Moores University, you do now finally have something to print out, and tack it to every idiots forehead who still insists that only half-marathon running, but not resistance training could save you from cardiovascular infarction ;-)

For their 6-months experiment, the scientists recruited 23 27±5 year-old healthy male subjects who were then randomized to either either endurance (ET, n=10) or resistance training (RT, n=13) in order to evaluate the long-term effects of these training modalities on brachial, femoral and carotid artery diameter and wall thickness (IMT), as well as femoral and brachial flow-mediated (FMD) and glyceryl trinitrate (GTN) mediated dilation. Ha? Well, let's say they wanted to know how the different exercise protocols, the scientists desribe as follows,

For those of you who want to add something to their regimen that boosts the longevity sirtuin, sirt-3 Carl and I have talked about during yesterday's show, here is the HMB + Leucine study I mentioned.

"[...] the ET intervention consisted of a progressivelyoverloaded programme of walking, jogging and running, inclusive of specified training phases over the 24week period. The focus of the periodized RT programme was Olympic weightlifting with incorporated supplemental exercises (e.g. dead-lift, back squat, front squat, bench press and overhead press) to develop overall strength and technique. Relative intensities for the ET and RT interventions were monitored throughout the sessions, individualized and progressed to ensure that subjects were exercising at prescribed percentages of VO2peak and 1RM, respectively" (Spence. 2013)

would effects the structure and performance of the hearts of their subjects, who completed three 1h training sessions of the respective exercise type per week.

As even resistance training enthusiasts should have expected, the RT group did not see significant improvements in O2Peak (ET yielded +5% increase in this indicator of cardiovascular fitness). On the other hand, the endurance training had no effect on upper body strength, which increased by statistically highly significant +21% in the resistance training group. Both these improvements are yet not really newsworthy and the whole study would not have made it to the SuppVersity Science Round-Up, respectively the Seconds, if it had not been for the high-resolution ultrasound images of the heart, which revealed:

Figure 3: Changes in body composition
relative to pre values (Spence. 2013)

• Resistance training affects the brachial artery, increases brachial artery resting (+8%, P<0.05), peak FMD  and GTN-mediated (P<0.01) diameters
• Endurance training affects the femoral artery, increases resting (+3%, P<0.05), peak FMD femoral artery diameters, and improvements in the femoral FMD-to-GTN% ratio 
• Both forms of training had similar beneficial effects the carotid artery wall thickness

In summary: Despite the fact that the two training forms will have differential effects on your overall conditioning and strength, they both increase lean mass (see figure 3) and decrease the artery wall thickness of the artery which supplies your brain with oxygen (the carotid artery). And with their unique effects on the brachial artery in the upper arm (RT) and the femoral artery in the tigh (ET), they make a perfect team to protect you from cardiovascular disease and stroke.

"I'd like a Levitra(R) enhanced green tea with probiotics..." What?

The last two items from yesterday's list that did not make it into the show are actually not brand new studies. One is a comment from the Journal of Clinical Investigation and the other one is a study that has been around since 2011, already. I picked them up in the course of the week and found them news-worthy, although it is not really sure, a combination of green tea, viagra and rothia bacteria is going to prevent cancer and solve your problems with gluten or whether one or the other or a combination of all is going to kill you ;-) Anyways, here are the details:

• 

  Rothia has hitherto rather been known as a bad guy, which can, if it gets into the wrong places cause endocarditis (heart), meningitis (brain), peritonitis (an inflammation of the peritoneum, the thin tissue that lines the inner wall of the abdomen and covers most of the abdominal organs) and other infections (Photo by Nathan Reading)

  Rothia bacteria in the upper gastrointestinal tract degrade gluten (Zamakharchi. 2011) -- Back in 2011 already a group of researchers from the Boston University Henry M. Goldman School of Dental Medicine published a paper in which they describe the astonishing ability of a common gram positive bacteria called Rothia, which is also present in the oral cavity could degrade 50% of the glyiadin peptides the scientists had added to the petri dish before the they were incubated for 30min.
  
  In view of the fact that the immunogenic parts of the peptides, that cannot be cleaved by our natural digestive enzymes, it does appear like a too happy coincidence that our microbial subtenants from the oral and upper gastrointestinal tract possess the enzymatic machinery to degrade the harmful gluten peptides for us - don't you think so?
  
  One thing is for sure, the presence of a bacteria that deals with the stuff, we cannot deal with on our own in our oral cavity certainly does support what I said in the context of the "bulimia apparatus" - the digestive process with all it's downstream effects on your metabolism starts in the mouth:
  

  "During mastication (chewing) foods are mixed with whole saliva helping to accelerate the break-down by digestive enzymes during the residency time in the oral cavity. Oral microorganisms in the swallowed food bolus may or may not survive and/or continue to exert proteolytic activities during or after gastric passage. Our in vitro data with R. aeria show that its enzymes are not abolished at acidic pH values, and are optimally active under more basic pH conditions. In vivo, this could mean that during gastric passage the enzymes will neither be active nor destroyed, and that enzymatic reactivation would occur upon transfer to the duodenum." (Zamakharchi. 2011)

  Aside from the fact that they could make it into the large intestine, previous studies have already confirmed that R. mucilaginosa can even gain a foothold there (Ou. 2009), so that probiotic supplementation with Rothia mucilaginosa could in fact turn out to be a viable treatment / prevention strategy for celiac disease and other gluten related health problems.

• EGCG + PDE-5 = cancer killer (Yang. 2013) -- Cancer is like the Learnean Hydra from ancient Greek mythology (see image on the right). Whenever you cut off one of its heads it'll grow two new ones... well, i must admit the analogy isn't perfect, but it's still useful to understand why the PDE-5 inhibitor vardenafil worked wonders when it was co-administered with the green tea catechin EGCG.
  
  

  

  You really have to wonder why these ancient heroes who had the cunning to kill the Hydra did not even wear a pair of speedos, when they entered into it's watery realms ;-)

  Just like the many Greek warriors who died in their battle with the Hydra before it was finally slain by Heracles, EGCG is well capable of doing major harm to the cancer cells. Just like the snake-like beast the cancer cells have yet found a way to resist the onslaught. Obviously not by growing new heads, though, but rather by over-expressing PDE-5 in order to repress the EGCG increased cGMP activity that would otherwise initiate apoptosis, i.e. the natural self-destruction mechanism of a cell.
  
  So what could be more straight forward than using a PDE-5 inhibitor just like Iolaus, Heracles cousin, used a firebrand to scorch the neck stumps of the Hydra after each decapitation. Clever and effective, don't you think so?
  
  Well, there still is one downside: Just like the story about the Hydra is nothing but a myth, the studies Yang and Wang reference in their commentary in the latest edition of the Journal of Clinical Investigation are only in-vitro studies and whether drinking green tea and popping one of Bayer's Levitra(R) pills after the other, is going to prevent, let alone kill existing cancer, will still have to be elucidated.

That's if for today! And since I am a bit in a hurry, I will leave it up to you to check out the latest really short news on the SuppVersity Facebook wall. Contrary to the green tea + Levitra treated cancer cells, the facebook news are guaranteed to proliferate even before the official Saturday installment of On Short Notice is going to be posted in ~24h (in fact, it will first have to researched and written ;-)

References:

• Bounous G, Gold P. The biological activity of undenatured dietary whey proteins: role of glutathione. Clin Invest Med. 1991 Aug;14(4):296-309.
• Kerasioti E, Stagos D, Jamurtas A, Kiskini A, Koutedakis Y, Goutzourelas N, Pournaras S, Tsatsakis AM, Kouretas D. Anti-inflammatory effects of a special carbohydrate-whey protein cake after exhaustive cycling in humans. Food Chem Toxicol. 2013 Jan 26.  [epub ahead of print] 
• Ou G, Hedberg M, Horstedt P, Baranov V, Forsberg G, et al. Proximal small intestinal microbiota and identification of rod-shaped bacteria associated with childhood celiac disease. Am J Gastroenterol. 2009;104:3058–3067.
• Peng X, Xiong YL, Kong B. Antioxidant activity of peptide fractions from whey protein hydrolysates as measured by electron spin resonance. Food Chemistry. 2009; 113(1):196–201. 
• Spence AL, Carter HH, Naylor LH, Green D. A prospective randomised longitudinal study involving 6-months of endurance or resistance exercise on conduit artery adaptation in humans. J Physiol. 2013 Jan 28. [Epub ahead of print] 
• Yang CS, Wang H. Cancer therapy combination: green tea and a phosphodiesterase 5 inhibitor? J Clin Invest. 2013 Jan 25:1-3. [Epub ahead of print]
• Zamakhchari M, Wei G, Dewhirst F, Lee J, Schuppan D, Oppenheim FG, Helmerhorst EJ. Identification of Rothia bacteria as gluten-degrading natural colonizers of the upper gastro-intestinal tract. PLoS One. 2011;6(9):e24455.