Tonya Estherhuizen, biostatistician from the College of Health Sciences, for her help with the statistical analyses. Purpose: The purpose of this study was to determine whether administration of Traumeel® S five days before and three days after a 90-minute downhill treadmill run at 75% V02 peak significantly alters systemic markers of the inflammatory response.
Conclusions and recommendations 58
Appendices 71
Introduction
Introduction to the Problem
The Effect of Traumeel On Inflammatory Markers in the Blood
In studies performed on whole blood cultures, increases in the concentration of Transforming Growth Factor Beta (TGF-6), which is a cytokine synthesized by Type 3 T-Helper (Th3) lymphocytes, were reported after administration of Traumeel® S (Heine & Schmotz, 1998). To determine whether 90 minutes of downhill treadmill running at 75% VO2 peak results in an increase in neutrophils, muscle enzyme (creatine kinase; CK), stress hormone (Cortisol), type 1 T-helper (Thl) cytokine (IL- 12p70) ), type 2 T-helper (Th2) cytokine (IL-6, IL-10), chemokine (IL-8) and pro-inflammatory cytokine (TNF-a) concentrations in the circulation.
Hypotheses
Limitations and Scope of the study
Subjective perceptions of postexercise delayed-onset muscle soreness (DOMS) were also monitored during the 72 h following each exercise trial.
Eccentric Exercise and Muscle Damage
Abnormally high circulating levels of enzymes such as CK are therefore considered to reflect changes in the integrity of muscle fiber membranes, either due to damage to, or increased permeability of, the membrane to the enzyme (Eston et al, 1995). This facilitates an inflammatory response and results in the clearance of the cellular debris and subsequent healing of the tissue (Pedersen, 2000; Pedersen and Hoffman-Goetz, 2000; Pedersen et al, 2001).
Cytokines and Exercise
- Introduction
- Pro-Inflammatory cytokines
- Tumor Necrosis Factor Alpha (TNF-a)
- Interleukin lp (IL-lp)
- Interleukin 1-Receptor Antagonist (ILl-ra)
- Chemokines
- Interleukin-8
- Immunomodulatory Cytokines
- Interleukin-12
- Pro and Anti-inflammatory Cytokine Response to Eccentric Endurance Exercise
Several studies have found a slight increase in blood IL-8 levels after moderate-intensity exercise (Henson et al., 2000; Suzuki et al., 2002). These play a crucial role in the control and isolation of the inflammatory process (Bagby et al., 1996; Peters, 2004).
The Thl - Th2 Differentiation; Corresponding Cytokine Production And its Effects
- Introduction
- Regulation of the Thl - Th2 Differentiation
- The Thl - Th2 Shift Effect During Exercise
- The Role of Cortisol
It is therefore considered an anti-inflammatory cytokine that aids wound healing by limiting the inflammatory response (Bonecchi et al., 1998; Elenkov and Chrousos, 1999). Early secretion of EL-4 leads to polarization of Th cell differentiation towards Th2-like cells (Mosmann et al., 1986). The Th2 cell secretion of IL-4 and IL-10 also leads to suppression of Th1 responses by down-regulating the production of macrophage-derived IL-12 and inhibiting the differentiation of Th1-type cells (Mosmann et al., 1986) Kelso, 1995).
Cortisol is thought to function by inhibiting the production of IL-12 by antigen-presenting cells (Elenkov and Chrousos, 1999) and also by reducing the ability of T cells to respond to IL-12 (Franchimont et al. , 2000). These cells express more 62-adrenoreceptors on the surface compared to Thl cells (Pedersen and Hoffman-Goetz, 2000) which may explain that only the percentage of type 1 T cells correlated with epinephrine during exercise (Steensberg et al ., (2000) ) study.
The Repeated Bout Effect
Postexercise concentrations have been shown to increase by more than 50% after 60 min of downhill running (gradient -13.5%) at 75% V02max (Smith et al., personal correspondence), but. The exercise-induced increase in systemic cortisol concentration is a sequel to stimulation of the HPA axis and adrenal cortex by IL-6, and increased circulating cortisol concentration has been reported in and in the bone marrow, causing a further release of neutrophils into blood. current (Starsky et al., 2003; Pedersen and Hoffman-Goetz, 2000; Suzuki et al., 1999). The connective tissue theory predicts that after the first period of eccentric exercise, muscles are protected by a general increase in intramuscular connective tissue (Frieden and Lieber, 1992; McHugh et al., 1999; Frieden and Lieber, 2001; McHugh, 2003).
An increase in the content of connective tissue should be ensured by intermediate filaments in muscle fibers, which consist of proteins, desmin, vimentin and synemin. During this process, the muscle-tendon complex is thought to adapt its viscoelastic properties to absorb and transmit the forces generated during sudden and strong contractions (Yu et al., 2003a; 2003b). 1999) explain that the cellular theory proposes that muscle damage occurs as a result of irreversible sarcomere tension during eccentric contractions.
The Effect of Traumeel On Inflammatory Markers in the Blood
Lussignoli et ai, 1999 report that the anti-inflammatory effect of Traumeel®S is presumably the result of the activity of individual components on the different phases of inflammation. The results of their study suggested that the local administration of Traumeel®S reduced edema development, and this inhibition was similar to the effect of aspirin at a dose of 30 mg/kg in the same experimental model. In an adjuvant arthritis model, the intrapaw administration of Traumeel®S every other day led to a significant reduction of acute local inflammation without affecting the chronic arthritic process (Conforti et al., 1997).
1999) reported a series of experiments aimed at improving knowledge of the therapeutic action of Traumeel®S using an intrapaw injection of a small amount of homologous blood mimicking a traumatic extravasation of blood, a condition commonly treated with Traumeel®S. They also tested the activity of individual components of Traumeel®S on blood edema and of a combination containing only components with known anti-inflammatory activity. Although additional studies are needed to clarify the mode of action of Traumeel®S and demonstrate the causal relationship between the inhibition of pro-inflammatory cytokine/chemokine secretion in cell culture and the reported clinical effects of the preparation, this in vitro work of Porozov et. al. 2004) and Heine and Schmotz (1998) shed more light on a possible mechanism for the anti-inflammatory effects of Traumeel®S observed in clinical use.
Traumeel® S and Exercise Induced Muscle Inflammation
2004) conducted a study examining the effects of Traumeel S in vitro on the ability of resting and PHA-, PMA-, or TNF-α-activated human T cells, monocytes, and intestinal epithelial cells to express LL-ip, TNF to divorce. -a and IL-8 for a period of 24-72 hours. Heine and Schmotz (1998) also demonstrated that Traumeel®S in whole blood cultures had the effect of increasing the levels of Transforming Growth Factor (TGF-β), a Th3 cytokine that suppresses Th1 and Th2 cell activity and also described acts as an anti-inflammatory cytokine (Mosmann and Coffman, 1989; while non-steroidal anti-inflammatory drugs act by inhibiting the arachidonic acid pathway and thus the production of prostaglandins through inhibiting the cyclooxygenase and lipoxygenase enzymes (Lussignoli et al., 1999 ; Porozov et al., 2004 , Ganong, 2005), however, Traumeel®S has been shown to control the generation of reactive oxygen species (ROS) by activated neutrophils.
Therefore, increased neutrophil oxidative burst activity and lymphocyte function are possible with the ingestion of Traumeel, as the suppressive effect of high levels of ROS on the leukocytes would be neutralized by Traumeel®S (Conforti et al., 1997, Peters, 1997b). These results were consistent with those of Bohmer and Ambrus (1992) and Wright-Carpenter et al. 2004), who reported improved recovery times for the investigated injuries in their respective studies compared to placebo.
Conclusion
- Study design, ethical approval and insurance
- Sample
- Study Protocol
- Baseline Laboratory Testing
- Track testing
- Matching of subjects
- The Intervention and Trial Procedure
- Post-Trial Recovery
- Treatment of the blood samples
- Analysis of Blood Samples
- Determination of Serum CK and Cortisol Concentrations
- Determination of Plasma IL-6 concentration by sandwich ELISA
- Flow Cytometric Analysis of Cytokine Concentrations 1. Introduction
- Subjective Rating of Perceived Muscle Soreness
- Statistical Analyses
Moreover, the subjects who did not adhere to the methodology of the study were also excluded. Prior to beginning the study, subjects were required to undergo basic laboratory and field testing. The coding was only revealed to the researchers after the study was completed and a preliminary report of the results was made available to Heel (Pty).
Subjects were asked to return to the exercise laboratory at the same time in the morning on days following the trial to obtain follow-up blood samples and to record subjective ratings of delayed muscle soreness. This was done using a flow cytometer equipped with a 48-nm laser capable of detecting and distinguishing fluorescence emissions at 576 and 670 nm (eg BD FACScan™ or BD FACSCalibur™ instruments) and BD Cellquest™ software.
Results
- Subject Characteristics
- Training Status and Performance during Field Testing
- Neutrophil Count
- Cortisol
- DOMS
- Plasma TNFa concentrations
- Plasma IL-6 concentrations
- Plasma IL-8 concentrations
- Plasma IL-10 concentrations
- Plasma IL-12p70 concentrations
The TRAU and PLAC groups did not differ significantly in terms of mean HR max, RPE max, VE max or peak V02 (p > 0.05). Simple post hoc analyzes with contrast verified these findings (p > 0.05) at each of the time points. Downhill running resulted in a small (5-fold, 5.2-fold) but significant elevation in IL-6 (p < 0.05) when compared to pre-test values. The difference in exercise-induced change between the two groups was, however, not statistically significant {p > 0.05).
In the TRAU and PLAC groups, there was a significant increase in IL-8 (p < 0.05) when compared to pre-test values. The difference in exercise-induced change between the two groups was, however, not statistically significant (p > 0.05).
Discussion
- Pre-Trial Data
- Trial Performance
- Post-Trial Data
- Serum Creatine Kinase
- Neutrophil Counts
- Serum Cortisol
- Plasma Interleukin - 6
- Plasma Interleukin - 8
- Plasma Interleukin -10
- Plasma Interleukin - 12p70
Interestingly, despite consistently higher mean cortisol concentrations in the TRAU group across the 5 time points, the difference between the two groups was not statistically significant. The exercise-induced increase in IL-6 was, however, relatively modest compared to previous reports in the literature. Downhill running resulted in an approximately 9-fold increase in serum IL-8 concentrations in the TRAU group and the PLAC group.
However, the difference in the exercise-induced change between the two groups was not statistically significant (p > 0.05). In both groups, therefore, the downhill running protocol not only resulted in increased neutrophil concentrations in the blood, but also increased concentrations of the chemotactic cytokine, IL-8. In addition, the concentrations of cortisol, IL-6, IL-8 and IL-10 in the blood were not significantly higher in the TRAU group.
Annotation of genes with potential roles in the immune system: six new members of the IL-1 family.
APPENDIX A
I agree to participate in a study to be conducted by Edith Peters-Futre and colleagues from the Department of Physiology at the University of KwaZulu-Nalal Medical School. Sublingual ingestion of 1 x TRAUMEEL or placebo tablet 3 x per day 30 minutes before the meal with each meal (i.e. 3 x per day), for 5 days prior to the exercise test (to be carried out in the Exercise Laboratory, Nelson R Mandela School of Medicine I am aware that there may be some degree of discomfort when the blood is drawn and that the exercise tests will involve strenuous, prolonged exertion which may be associated with certain medical risks and result in muscle pain in the 48- 72 hours after the test.
I also understand that I have no recourse against the University in the event of an acute medical event that occurs while I am running on the treadmill. I understand that participation is completely voluntary and that I can withdraw from the study at any time.
APPENDIX C
