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Roeland Van Wijk and Fred AC Wiegant are from the he essence of homeopathy is the stimulation of bio-
Department of Molecular Cell Biology, Faculty of Biology,
Utrecht University, Utrecht, the Netherlands.
The Similia Principle as a Therapeutic Strategy ALTERNATIVE THERAPIES, MARCH 1997, VOL. 3, NO. 2 33
death. This article thus focuses on the research strategies for Major stress protein families and their functions
determining which cellular components play a role in defense
and on the integration of this information. The chosen research Hsp Stress protein Functions in cell metabolism
strategy is on complex systems, which requires the analysis of family and cellular stress defense
nonlinear interactions in a network. The task of discovering the
hsp100 hsp100 Heat and ethanol tolerance
nature of the defense process during interventions according to
the similia principle is the next topic of research. Next, the pos- hsp90 hsp90 (hsp84) Stabilization of proteins;
tulated, precise kinetic and interactive behavior of defense pro- maintenance of the inactive
teins must be tested and it must be shown that the results of the protein form during trans-
port
various experiments, when fitted into a quantitative model, are
in accord with the observed changes. Essential for cell viability
34 ALTERNATIVE THERAPIES, MARCH 1997, VOL. 3, NO. 2 The Similia Principle as a Therapeutic Strategy
Pattern of Protector Proteins is Damage Specific replenishment of these protector proteins starts with activation
A variety of different protector proteins have been found. of associated protector protein gene promoters on the cell’s
Interestingly, the pattern of protector proteins that is induced DNA. This highly specific activation occurs by binding of specif-
appears to be damage specific.8-10 Under different deleterious ic DNA-binding factors, called heat shock transcription factors
conditions, qualitatively different patterns of protector proteins (HSFs), on these DNA sites.11 The binding of the HSF to the pro-
seem to be induced. Although an explanation for such a complex moter on the cell’s DNA is the signal that triggers transfer of
regulation is lacking, the patterns do suggest some differentia- information from DNA into messenger RNA (mRNA), leading
tion in the induction of the type of damage-induced protection eventually to synthesis of new protector proteins. Whether or
mechanisms. not these DNA-binding factors interact with the DNA depends
Because these protector proteins are essential to survive on the existing quantity of protector proteins in the cell. The
threatening conditions and because their induction occurs at genome is specifically activated to trigger this synthesis of addi-
concentrations of various toxic compounds that are not yet tional protector proteins only when the quantity of protector
lethal for the cells, the damage-specific pattern of induction may proteins falls below a certain threshold. Normally, at least one
be considered as the cellular equivalent of the homeopathic rem- type of protective protein, hsp70, and a DNA-binding factor,
edy picture. HSF, form a complex that provides the basis for this regulation.
If protector proteins are required to neutralize abnormal pro-
The Problem of Regulation teins, this complex then dissociates, releasing HSF, which then
of Availability of Protector Proteins binds to the promoters and induces production of mRNA, with
A simple model for the regulation of protector proteins in the ensuing synthesis of new protector proteins. When sufficient
defense after cell damage is depicted in Figure 1. The quantity of new protector proteins have been produced, that is, when the
free protector proteins available in the cell decreases after cell level of these proteins is raised above the threshold value, hsp70
damage. As long as these protector proteins are available, dam- will again form a complex with HSF molecules, uncoupling the
age is reduced to a minimum. However, when a shortage of pro- HSF from DNA, with a concomitant halt in production of
tector proteins arises in the case of an overload of damage, the mRNA. In terms of systems theory, one might say that this is the
abnormal protein molecules can complex with other cell struc- autoregulation loop that forms the basis of damage-induced
tures. Cell damage and death can then be avoided only by pro- recovery processes.
duction of increasing amounts of new protector proteins. The
Mathematical Modeling as a Future Step
in Understanding the Regulation of Defense
Defense and recovery are not simple and singular phenom-
Proteotoxicity Compensation cycle ena but are determined by the kinetic parameters of the autoreg-
HSF (inactive) ulation loop as illustrated in Figure 2. Insight into the
effectiveness of this loop for cellular defense can therefore be
Pool of free obtained only from studies involving the parameters mentioned
HSPs in this mathematical model. This mathematical model of defense
Stress
Protector protein and recovery processes has only recently started with the devel-
opment of a model of hsp70 regulation in the cell.12 So far, five
main blocks, each describing one or more processes involved in
HSF (active) the functions of hsp70 and its synthesis, have been developed.1,12
Denatured These blocks constitute the basic structure of a mathematical
DNA
protein model for the regulation of hsp70 in cells. The major processes
included in the model so far are (1) denaturation of proteins
HSPmRNA after an increase in temperature and the binding of hsp70 to
denatured or nascent proteins; (2) interaction of hsp70 with HSF
and the activation of HSF not bound to hsp70; (3) interaction of
activated HSF with a specific HSF binding sequence (heat shock
element, HSE) on DNA; (4) the relationship between HSE and
HSP transcriptionally active HSF and the level of hsp70mRNA active-
ly engaged in synthesis of hsp70; and (5) translation of viable
FIGURE 1 Basic processes of damage and defense at the cellular hsp70mRNA into its protein.
level. Proteotoxicity is illustrated as a change in protein conforma-
In the model constructed so far, the output of this regula-
tion. Protector proteins interact with these abnormal proteins.
After depletion of the pool of free HSPs, supplementation of these tion is the level of free, that is, unbound, hsp70 in the cell. Future
protector proteins occurs by way of the compensation cycle. steps in developing this mathematical model concern the inclu-
sion of lethality in order to predict stress-dependent survival and
The Similia Principle as a Therapeutic Strategy ALTERNATIVE THERAPIES, MARCH 1997, VOL. 3, NO. 2 35
SIMILIA PRINCIPLE
2.0 A According to homeopathic insights, recovery can be stimu-
lated by all kinds of substances, provided that they are applied in
a specific way. Of all substances, the one most suitable for stimu-
Relative synthesis of hsp70
1.5 lation of recovery is the one that can produce the artificial situa-
tion of disorder that most closely resembles the disordered state.
In other words, this concept hinges on the resemblance between
1.0 symptoms of the disturbed system and the symptoms caused by
the applied substance in a healthy system.
40
has been studied by a step-down heating protocol in which the
initial treatment with high heat is immediately followed by a sec-
ond treatment at lower doses. When these step-down heating
conditions were used, an enhanced synthesis of protector pro-
20 teins was indeed seen.13-15 Other studies show an enhanced occur-
rence of thermotolerance when mild step-down heatings were
used.16
The next relevant question is whether this response is a gen-
0
C x X X-x eral occurrence and can thus also be observed after stressor con-
ditions other than heat shocks. To this end, a damage-inducing
treatment of cells with arsenite or cadmium was followed by low
FIGURE 2 Schematic presentation of the effect of a low dose of a doses of arsenite or cadmium, respectively. When this step-down
homologous stressor on the synthesis of hsp70 (A) and on devel- treatment with arsenite or cadmium was used, cells exhibited an
opment of tolerance (B). When the stress condition is followed
enhanced synthesis of various protector proteins and develop-
by a low dose of the same stressor, an enhancement of hsp70 syn-
thesis (filled triangles) and of tolerance development is observed ment of tolerance.17,18 The lower doses had no effect on synthesis
in comparison with the effect of the stress condition alone (open of protector proteins or development of tolerance in cells that
circles). A low dose of the stressor does not influence the control were not pretreated (ie, healthy cells).
cells (open triangles). C=control, x=low-dose stressor, X=stres- Figure 2 shows, schematically, the transient induction of
sor, X-x=stressor followed by low-dose stressor. hsp70. The synthesis of this protein is further enhanced when a
low stressor dose is applied during the so-called posttreatment
period. The level of tolerance that is achieved is also higher when
the low dose is applied after treatment as compared to cells that
changes in cellular sensitivity to stressors. received a pretreatment only.
To sum up, the integrity of the cellular systems, their
defense and recovery, depends not only on the available protec- The Specificity of the Low-Dose Effect
tive proteins but also on the speed of activation of the total res- With respect to the specificity of the defense-enhancing
cue mechanism. Future developments in this research are effect of low doses of an analogous stressor, some recent studies
directed at the damage dependency and differentiation of the examined induction of protector proteins in sensitized cells by
mentioned production process. A quantitative experimental low doses of analogous but potentially damaging conditions. So
analysis and a mathematical evaluation are considered essential far, experiments have examined the effects of pretreatments
for this study and for understanding any interventions in this given with either a heat shock, sodium arsenite, or cadmium
production process, particularly interventions according to the chloride. After these pretreatments, the cultures were exposed to
similia principle. low doses of heat, arsenite, or cadmium or to control conditions.
36 ALTERNATIVE THERAPIES, MARCH 1997, VOL. 3, NO. 2 The Similia Principle as a Therapeutic Strategy
The stimulating effect on the synthesis of various protector pro-
teins was then studied. This application of either heat shock, 6 6
arsenite, or cadmium at subliminal conditions to already disor-
Heterologous Reinduction
Recently, the specificity of desensitization of protector pro-
1.0 tein induction was tested with heat shock, sodium arsenite, and
cadmium chloride used as primary and secondary inducers of
protector proteins.23 The data suggest that the degree, but not
the pattern, of reinduction of protector proteins is influenced by
the type of stressor used in the pretreatments. Thus, stimulation
of synthesis of protector proteins after cadmium as the sec-
0 ondary stressor is severely inhibited in cadmium-pretreated
After 0 as cd 0 as cd cells, whereas reinductions after arsenite and heat shock are
Before As Cd specifically inhibited in cells pretreated with arsenite and heat
shock, respectively.
In summary, homologous sensitization is transient, and
FIGURE 3 Schematic presentation of the specificity in stimula- homologous desensitization develops specifically. Apparently, a
tion of HSP synthesis by low doses of stressors. A high dose common denominator regulates the coordinate expression of a
induces a certain level of HSPs, which is further enhanced when group of protector proteins. Heterologous reinduction can still
subsequently incubated with a low dose of the same stressor, but
occur but, in that case, the pattern of protector proteins induced
not with a low dose of stressor conditions that were not similar.
A low dose exerts no effects on the level of hsp70 found in con-
by the secondarily applied stressor shows a stressor specificity
trol conditions (black bars). that is dependent only on the second treatment and is indepen-
dent of any pretreatment.
The Similia Principle as a Therapeutic Strategy ALTERNATIVE THERAPIES, MARCH 1997, VOL. 3, NO. 2 37
Future Research Aimed application of low doses of damaging conditions. The selection
at Further Understanding of the Similia Principle of condition can be based on the molecular symptoms of the cel-
The possible implication of the results mentioned so far is lular defense process. In this respect, the application of low
that the increased production of protector proteins at a later doses according to the similia principle warrants further study.
time after an injury, that is, during the period of homologous tol- It should now also be easier to understand the role of these
erance, occurs only after application of a heterologous damaging cellular recovery processes in an organ’s functionality after expo-
compound. When we consider the pattern of synthesized protec- sure of an intact organism to stressful conditions. Therefore, the
tor proteins as the molecular symptoms of recovery, the speci- similia principle can be expected to manifest itself as a general
ficity of the similia principle reveals itself in the degree of biological phenomenon. This offers exciting opportunities for
relatedness between the effects of the disturbing compound on developing new avenues for therapeutic interventions in bio-
the pattern of synthesis of these protector proteins. We must medicine.
search for analogous damaging conditions that induce patterns
of protector proteins similar to those generated by the primary Acknowledgments
C A van der Mast, PhD, is gratefully acknowledged for critical reading of the manuscript. This work
stressor without being identical to this primary stressor. For this was supported by the HomInt organization, Karlsruhe, Germany.
step, different compounds must be tested at the cellular level.
Another possibility is to look for conditions that induce the References
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38 ALTERNATIVE THERAPIES, MARCH 1997, VOL. 3, NO. 2 The Similia Principle as a Therapeutic Strategy