Research Update #16

by Hypertrophy

As we approach the new millennium we find the science of building muscle progressing faster than ever before. Long gone are the days of simple trial and error when it comes to building muscle. The modern bodybuilder demands more than just "hear say" if they are to adopt a new training routine or nutritional supplement. This column was created to keep today’s bodybuilder on the cutting edge of scientific research that might benefit them in their quest for body perfection.

Are steroids your key to preventing Andropause?

Title: Long-term suppression of leydig cell steroidogenesis prevents leydig cell aging.

Researchers: Chen H, Zirkin BR

Division of Reproductive Biology, Department of Biochemistry and Molecular Biology, Johns Hopkins University, School of Hygiene and Public Health, 615 North Wolfe Street, Baltimore, MD 21205, USA.

Source: Proc Natl Acad Sci U S A 1999 Dec 21;96(26):14877-81

Based on the observations that reactive oxygen is capable of damaging components of the steroidogenic pathway and that reactive oxygen is produced during steroidogenesis itself, we hypothesized that long-term suppression of steroidogenesis might inhibit or prevent age-related deficits in Leydig cell testosterone production. To test this, we administered contraceptive doses of testosterone to groups of young (3 months old) and middle-aged (13 months old) Brown Norway rats via Silastic implants to suppress endogenous Leydig cell testosterone production. After 8 months, the implants were removed, which rapidly (days) restores the ability of the previously suppressed Leydig cells to produce testosterone. Two months after removing the implants, when the rats of the two groups were 13 and 23 months of age, respectively, the Leydig cells in both cases were found to produce testosterone at the high levels of young Leydig cells, whereas significantly lower levels were produced by the 23-month-old controls. Thus, by placing the Leydig cells in a state of steroidogenic "hibernation," the reductions in Leydig cell testosterone production that invariably accompany aging did not occur. If hormonal contraception in the human functions the same way, the adverse consequences of reduced testosterone in later life (osteoporosis, reduced muscle mass, reduced libido, mood swings, etc.) might be delayed or prevented.

Discussion:

OK, I will admit that I am biased about the use of androgens. Frankly, I think politicians and the popular press have unjustly vilified them. This being the case I tend to take notice to studies involving androgens. This particular study was a real surprise.

Male aging is accompanied by reduced testosterone production by the Leydig cells, the testosterone-producing cells of the testis. The mechanism by which this occurs is unknown. In the above study they suppressed testosterone production with androgen implants in male rats for eight months. The researchers called this behavior of the testes as "steroidogenic hibernation".

This is the same phenomenon that all male bodybuilders experience when using testosterone of other androgens. After removing the hormone implants they measured the ability of the leydig cells to produce testosterone by bathing them in luteinizing hormone (LH). They measured testosterone production as well as steroidogenic enzyme activities. As you can see from the graph above, the leydig cells of old rats that had their testosterone production temporarily shut down, retained their ability to produce testosterone equal to young rats. The activities of each enzyme tested were significantly lower in Leydig cells from the 23-month-old control rats compared with the 13-month-old control rats. All the steroidogenic enzyme activities were reduced in Leydig cells isolated from 11- and 21-month-old rats tight after exogenous testosterone treatment for 8 months. Two months after the implants were removed, when the rats were 13 and 23 months of age, respectively, the activities of each of the suppressed enzymes were at the high levels of 13-month-old control rats, in each case significantly higher than those of the 23-month-old controls.

The researchers hypothesize that this phenomenon is the result of decreased oxidative damage to leydig cells. In the course of normal cell function, reactive oxygen species are produced and can cause damage to DNA, proteins (e.g. enzymes), and cell membranes including mitochondrial membranes. This could contribute to age related loss of testosterone production. It is known that reactive oxygen species can damage the steroidogenic pathway in vitro (Quinn, 1985).

The authors of this study considered the dose of testosterone needed to accomplish this as a "contraceptive" dose. In men this is about 100 mg testosterone enanthate per week. They will usually throw in some progesterone too. This is far below what you would need to be a competitive bodybuilder. It could be that in doses used by larger bodybuilders is too much and may actually cause permanent losses in testosterone production. On the other hand, these doses might be equally effective or even more so than contraceptive doses. Time will tell. In any case, a preservation of testosterone production in later life means decreased risk of osteoporosis, sarcopenia, loss of strength, mood swings, etc. Appropriate studies carried out on humans is clearly needed.

Title: Effects of anabolic steroids on the muscle cells of strength-trained athletes.

Researchers: Kadi F, Eriksson A, Holmner S, Thornell LE Department of Integrative Medical Biology, Umea University, Sweden.

Source: Med Sci Sports Exerc 1999 Nov;31(11):1528-34

Athletes who use anabolic steroids get larger and stronger muscles. How this is reflected at the level of the muscle fibers has not yet been established and was the topic of this investigation. METHODS: Muscle biopsies were obtained from the trapezius muscles of high-level power lifters who have reported the use of anabolic steroids in high doses for several years and from high-level power lifters who have never used these drugs. Enzyme-immunohistochemical investigation was performed to assess muscle fiber types, fiber area, myonuclear number, frequency of satellite cells, and fibers expressing developmental protein isoforms.

RESULTS: The overall muscle fiber composition was the same in both groups. The mean area for each fiber type in the reported steroid users was larger than that in the nonsteroid users (P < 0.05). The number of myonuclei and the proportion of central nuclei were also significantly higher in the reported steroid users (P < 0.05). Likewise, the frequency of fibers expressing developmental protein isoforms was significantly higher in the reported steroid users group (P < 0.05).

CONCLUSION: Intake of anabolic steroids and strength-training induce an increase in muscle size by both hypertrophy and the formation of new muscle fibers. We propose that activation of satellite cells is a key process and is enhanced by the steroid use. The incorporation of the satellite cells into preexisting fibers to maintain a constant nuclear to cytoplasmic ratio seems to be a fundamental mechanism for muscle fiber growth. Although all the subjects in this study have the same level of performance, the possibility of genetic differences between the two groups cannot be completely excluded.

This study was trying to answer one basic question, "How do anabolic steroids produce muscle growth?" If you were to ask the average bodybuilding enthusiast I think you would hear, "they increase protein synthesis." This is not untrue, its just that it is only part of the answer. In fact, the answer must include virtually every mechanism involved in skeletal muscle hypertrophy. These include:

• Enhanced growth factor activity (e.g. GH, IGF-1, etc.)
• Enhanced activation of myogenic stem cells (i.e. satellite cells)
• Enhanced myonuclear number (to maintain nuclear to cytoplasmic ratio)
• Enhanced protein synthesis
• New myofiber formation

You can read more about these mechanisms elsewhere in Mesomorphosis . Now, where do androgens fit in to this? Starting with growth factor activity, we know that testosterone increases GH and IGF-1 levels. In a recent study by Fryburg (Fryburg, 1997) the effects of testosterone and stanozolol were compared for their effects on stimulating GH release. Testosterone enanthate (only 3 mg per kg per week) increased GH levels by 22% and IGF-1 levels by 21% whereas oral stanozolol (0.1mg per kg per day) had no effect whatsoever on GH or IGF-1 levels. This study was only 2-3 weeks long, and although stanozolol did not effect GH or IGF-1 levels, it had a similar effect on urinary nitrogen levels. What does this difference between testosterone and stanozolol mean? In my opinion it means that stanozolol can increase protein synthesis by binding to AR receptors in existing myonuclei, however, because it does not increase growth factor levels it is ineffective at activating satellite cells and therefor may not increase satellite cell activity and thus myonuclear number.

Enhanced activation of satellite cells requires IGF-1. Those androgens that aromatize are effective at not only increasing IGF-1 levels but also the sensitivity of satellite cells to growth factors (Thompson, 1989). This action has no direct effect on protein synthesis. It leads to a greater capacity for protein synthesis by increasing fusion of satellite cells to existing fibers. This increases the number of myonuclei and therefore the capacity of the cell to produce proteins.

So it is not only that testosterone increases protein synthesis by activating genetic expression, it also increases the capacity of the muscle to grow in the future by leading to the accumulation of myonuclei which are required for protein synthesis. There is good reason to believe that testosterone in high enough doses may even encourage new fiber formation. To quote the authors of this study:

"Intake of anabolic steroids and strength-training induce an increase in muscle size by both hypertrophy and the formation of new muscle fibers. We propose that activation of satellite cells is a key process and is enhanced by the steroid use."

Just a note for those of you who will use androgens but refuse to use supplements. There is good evidence that creatine specifically stimulates the formation of myofibrillar protein in newly developing fibers (Ingwall, 1972; 1974; 1975; 1976). The effect is concentration dependent, maxing out at about 250 µM. Normal plasma concentrations are about 100 µM. In my opinion, creatine supplementation is certainly called for in athletes using heavy androgens.

A separate argument related to this study

I would like to quickly address a separate issue related to steroid use. Although I believe Bill Roberts has competently addressed this issue previously in earlier issues of Mesomorphosis, I would like to make my own contribution here. It has been thought, and is still commonly believed, that using steroids decreases the number of "steroid" receptors. This argument is used to explain the fact that growth eventually stops while using a given amount of steroid. Once you understand all of the effects of testosterone on growth factor levels and muscle cells you come to realize that the opposite is in fact the case. Simply stated, supraphysiological levels of testosterone gives rise to increased numbers of myonuclei and thereby an increase in the number of total androgen receptors per muscle fiber. Therefor, the larger you get from using steroids, the more receptive your muscle become to the presence of testosterone. Keep in mind that I am referring to testosterone and testosterone esters. Not the neutered designer androgens that people take to avoid side effects (Fryburg, 1997). This is not an argument to rapidly increase the dosages you use. It takes time for these changes to occur and the benefits of higher testosterone levels will not be immediately realized.

Ingwall JS, Morales MF, Stockdale FE, Wildenthal K. Creatine: a possible stimulus skeletal cardiac muscle hypertrophy. Recent Adv Stud Cardiac Struct Metab 1975;8:467-81

Ingwall JS, Morales MF, Stockdale FE. Creatine and the control of myosin synthesis in differentiating skeletal muscle. Proc Natl Acad Sci U S A 1972 Aug;69(8):2250-3

Ingwall JS, Weiner CD, Morales MF, Davis E, Stockdale FE. Specificity of creatine in the control of muscle protein synthesis. J Cell Biol 1974 Jul;62(1):145-51

Ingwall JS, Wildenthal K. Role of creatine in the regulation of cardiac protein synthesis. J Cell Biol 1976 Jan;68(1):159-63

Even pathetic training routines can cause hyperplasia in untrained women.

Title: Training affects myosin heavy chain phenotype in the trapezius muscle of women.

Researchers: Kadi F, Thornell LE.   Department of Anatomy, Umea University, Sweden.

Source: Histochem Cell Biol 1999 Jul;112(1):73-8

The aim of this investigation was to determine whether 10 weeks of three different types of training could alter the myosin heavy chain (MyHC) composition of the trapezius muscle. Twenty-one women were randomly assigned to three training groups that performed strength (n=9), endurance (n=7) or coordination training (n=5). Pre and post biopsies were taken from the upper part of the descending trapezius muscle and were analyzed for MyHC isoform content using 5% gel electrophoresis. In addition, we have studied the expression of embryonic and neonatal MyHCs using double-immunofluorescence staining. In the strength-trained group, there was a significant increase in the amount of MyHC IIA and a significant decrease in the amount of MyHC IIB and MyHC I. In the endurance group, there was a significant decrease in the amount of MyHC IIB. MyHC composition in the coordination group was not altered. Following the training period, myotubes and individual small-sized muscle fibers were observed in the strength and endurance trained groups. These structures were stained with the markers for early myogenesis (MyHC embryonic and neonatal). These data suggest that specific shifts in MyHC isoforms occur in the trapezius muscle following strength and endurance training. The presence of small-sized muscle fibers expressing the developmental isoforms of MyHC suggests that strength and endurance training induced the formation of new muscle fibers.

Discussion:

The title of this study doesn’t immediately grab your attention, however, after reading the results we are witness to a paradigm shift in thought. Let me clarify by stating that this paradigm shift is occurring in the ivory towers of academia, not in the more progressive thinking minds in bodybuilding. This shift is occurring to accommodate the fact that human adult muscle tissue does experience hyperplasia.

This is not the first study to shown the formation of new muscle fibers as a result of training. Even this same group has shown hyperplasia to occur in power lifters (Kadi, 1999). However, this is the only one I have came across involving previously untrained women.

In this study, groups of women participated in rather mild exercise programs. One group lifted weights (3X10-12 reps), another group used an upper body cycle ergometer (4X3 minutes) combined with very easy arm exercises with elastic bands (30 reps). The final group performed yoga-like exercises emphasizing balance and serenity with insignificant (I assume without weights of any kind) resistance during movements. Following the training period, newly formed myotubes and individual small-sized muscle fibers expressing embryonic and neonatal proteins were observed in both the strength and endurance trained groups. The amount of these small fibers producing neonatal and embryonic proteins was small (~3%) nevertheless, they were clearly a result of the exercise. Previous investigations of power lifters found the same relative proportion of newly forming fibers as a result of training (Kadi, 1999).

The power lifters in a subsequent study by Kadi were highly competitive and participated in Swedish national and international competitions. This is in stark contrast to the study above. The athletes in this later study participated regularly in international competitions (World Champion-ships, European Championships and Swedish Championships in power events). The athletes trained regularly for 2–3 hours, 4–6 times a week. The sessions consisted of 4–7 sets of exercises that were repeated 3–12 times per session. These athletes claimed to have never used anabolic steroids. They were selected from the same club where they signed a contract that committed them never to use any drug. Four of them participated voluntarily as controls for another project aiming to find more effective methods to detect drugs. As in this study, they found an average of ~3% small fibers producing neonatal and embryonic proteins.

As I have predicted in the past, future bodybuilding training routines will focus on producing damage instead of strength or fatigue. Of course the later two are inevitable consequences of training, nevertheless, efficiency will depend on narrowing our focus on the true stimulus of muscle growth, namely fiber microtrauma and subsequent activation of satellite cells. I currently have many of my clients using just such a routine training every body part three times per week, focusing on load (more weight, more stretch, more eccentrics, more frequency, more soreness.) instead of high volume, waiting for complete recovery, "strength endurance" training (my description of how most natural bodybuilders train). Soft tissue massage is also an integral yet difficult to obtain factor which enhances activation of satellite cells. Early results are promising. All (both men and women) who are using my system are sore almost continuously despite minimal training volume and are making new personal records in both size and strength.

Kadi F · Anders Eriksson · Staffan Holmner Gillian S. Butler-Browne · Lars-Eric ThornellCellular adaptation of the trapezius muscle in strength-trained athletes. Histochem Cell Biol (1999) 111:189–195