Phyto-adaptogens protect against environmental stress-induced death of embryos from the freshwater snail Lymnaea stagnalis

doi:10.1016/S0944-7113(00)80060-4

Phytomedicine

Volume 7, Issue 5, October 2000, Pages 389-399

https://doi.org/10.1016/S0944-7113(00)80060-4 Get rights and content

Summary

The main purpose of the studies presented in this paper is twofold: 1) to evaluate whether phyto-adaptogens (Acanthopanax senticosus and Rhodiola rosea) are able to exert a protective action against stress-induced death of embryos of the pond snail Lymnaea stagnalis; and 2) whether a possible protective action by phyto-adaptogens can be explained by the induction of heat shock proteins.

Enhancement in resistance by phyto-adaptogens was studied by applying plant extracts for a period of 20 hours to 3-day old larvae of the pond snail Lymnaea stagnalis. Subsequently they were exposed to a high and toxic dose of different environmental stressors. The following stress conditions were selected: a physical stress condition (heat shock: 43°C for 4 minutes), an oxidative stress condition (superoxide radicals induced by menadione (600μM for 2 hours)) and heavy metal-induced stress (copper (150μM for 1 hour) or cadmium (20μM during 1 hour)).

Both Acanthopanax and Rhodiola exert a strong protective action against a lethal heat shock. These adaptogens also significantly protect against the negative effect of superoxide radicals as induced by menadione. With respect to the protective action against exposure to heavy metals a small but significant protection was observed against intoxication with copper or cadmium by the phyto-adaptogens. In summary, there appears to be a difference in efficiency in enhancing resistance to the various stress conditions used (heat shock>menadione>copper>cadmium). Based on the results presented in this paper, we can conclude that phyto-adaptogens are able to enhance the resistance against the different stress conditions tested in developing individuals of Lymnaea. Although the degree to which resistance is enhanced appears to depend on the type of stressor applied, our results confirm the definition of phyto-adaptogens as being universal enhancers of non-specific resistance against different kinds of stress conditions.

With respect to the mechanism of enhanced resistance, the question was asked whether this protective action is caused by an induction of heat shock proteins (hsps), which are known to be involved in tolerance and adaptation. The phyto-adaptogens did not induce the synthesis of any of the hsps, nor did they modulate the normal heat shock induced synthesis of these stress proteins. We conclude that it is unlikely that hsps play a major role in obtaining an enhanced state of resistance provided by phyto-adaptogens.

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During the golden age of research of adaptogens, Brekhman [20] outlined the term adaptogen as: Non-specific action: This provides stimulance against physical, chemical or biological lethal agents. The non-specific protective outcome of phytoadaptogens enhanced the resistance of Lymnaea stagnalis larvae irrespective of stress conditions like: (i) Physical stress: heat shock of 43 °C for 4 min (ii) Oxidative stress: superoxide radicals induced stress by menadione at concentration of 600 μM for 2 h and iii) Heavy metal stress: copper induced stress at concentration of 150 μM for 1 h and cadmium induced stress at concentration of 20 μM for 1 h. Furthermore, the observed preventive action against environmental induced stress conditions appeared to be independent of nature of extract (aqueous or ethanolic) [21]. Similarly, in humans the immunobiological screening of a phytoadaptogenic preparation exerted non-specific immunomodulatory and interferonogenic potential on immunological phenotype of lymphocytes of patients suffering with ovarian cancer at different stages [22].
Natural products are of great surge in the identification of chemopreventive agents and biologically active molecules for the development of new promising therapeutic agents. These agents influence the cascade of biochemical and molecular signalling pathways involved in numerous physiological and pathological processes. The natural agents combat the dogma associated with the most dreaded, unconquered health concern and a multigenic disease- cancer. A category of plants known as adaptogens maintain perturbed homoeostasis, augment adaptations to noxious stimuli (exposure to cold, heat, pain, general stress, infectious organisms) and offer endurance to attenuate several disorders in human beings. The well known adaptogens and immunomodulators such as Rhodiola rosea, Withania somnifera, Tinospora cordifolia, Bacopa monnieri, Emblica officinalis, Glycyrrhiza glabra, Asparagus racemosus, Ocimum sanctum and Panax notoginseng claimed to have significant antioxidant and anticarcinogenic properties due to the presence of various biologically active chemical compounds. Their immunopotentiating activity is mediated through the modulation of T-cell immunity biochemical factors, transcription factors, some genes and factors associated with tumor development and progression. The combinatory formulation of active immunostimulating constituents from these plants may provide better homeostasis. These immunostimulant factors suggest their potential therapeutic significance in adjuvant or supportive therapy in cancer treatment.
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While most conventional antidepressant drugs for depression modulate brain monoamine and indolamine activity (Nutt 2008) recent data suggest that the antidepressant effect of R. rosea may be associated with key mediators of stress response, regulation of homeostasis of HPA axis activity (Fig. 2), modulation of G-protein coupled receptor (GPCR) signaling pathways (Fig. 3) and other molecular networks involved in depression (Panossian et al. 2012, 2013, 2014) (Tables 1–4). Mechanisms of action of R. rosea extract (and its active constituents) were studied in experiments on isolated cells (Panossian et al. 2012, 2013, 2014; Wiegant et al. 2008; Boon-Niermeijer et al. 2000; Schriner et al. 2009), nematodes (Wiegant et al. 2009), laboratory animals (Panossian et al. 2007, 2008a, 2008b; Panossian and Wagner 2005; Prodius et al. 1997) and humans (Olsson et al. 2009). Another putative antidepressant mechanism of R. rosea extract may be its effect on neuropeptide-Y (NPY) mediated upregulation of heat shock protein Hsp-70 which, in turn, down-regulates stress-induced JNK protein (suppressing glucocorticoid receptors and increasing cortisol) (Panossian et al. 2007) (Fig. 2).
Rhodiola rosea (R. rosea) is a botanical adaptogen with putative anti-stress and antidepressant properties. Evidence-based data supporting the effectiveness of R. rosea for depression in adults is limited, and therefore a comprehensive review of available animal and human studies suggesting a putative antidepressant action is warranted.
A review of the literature was undertaken to ascertain studies of possible antidepressant mechanisms of action and studies of the safety and effectiveness of R. rosea extracts in animals and adult humans.
A search of MEDLINE and the Russian state library database was conducted (up to October 2015) on R. rosea.
Mechanism of action: R. rosea extracts and its purified constituent, salidroside, has been shown to produce a variety of mediator interactions with several molecular networks of neuroendocrine-immune and neurotransmitter receptor systems likely to be involved in the pathophysiology of depression. A wide variety of preclinical in vivo and ex vivo studies with laboratory animals suggests the presence of several biochemical and pharmacological antidepressant-like actions.
Clinical assessment of R. rosea L. rhizome extracts in humans with various depressive syndromes is based upon results from two randomized, double-blind, placebo-controlled trials of 146 subjects with major depressive disorder and seven open-label studies totaling 714 individuals with stress-induced mild depression (diagnosed as asthenic syndrome or psychoneurosis). Overall, results of these studies suggests a possible antidepressant action for R. rosea extract in adult humans.
In contrast to most conventional antidepressants, R. rosea extract appears to be well-tolerated in short-term studies with a favorable safety profile.
R. rosea demonstrates multi-target effects on various levels of the regulation of cell response to stress, affecting various components of the neuroendocrine, neurotransmitter receptor and molecular networks associated with possible beneficial effects on mood.
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Adaptogens are the plant-derived biologically active substances that appear to induce a state of non-specific increase of resistance of the organism to diverse assaults that threaten internal homeostasis and improve physical endurance. The pharmacological assessment of adaptogens typically includes evaluation of their stimulating, tonic and stress protective effects in different screening models in which animals are challenged to acute and chronic stress conditions. Another clue to the identity of plant adaptogens are the chemical compounds found in them that may be responsible for the adaptogenic properties. Adaptogenic activity was seen in withania and rhodiola herbs. Mechanisms of action to improve adaptation to stress were as follows: W. somnifera demonstrated activity on the HPA axis, on the CNS and also had an antioxidant action Rhodiola species demonstrated possible CNS activity, antioxidant and anti-inflammatory actions.
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Exposing isolated Lymnaea stagnalis eggs individually proves to be a suitable method for generating chronic toxicity endpoints on snail development. Embryo toxicity tests with L. stagnalis have previously been conducted by Boon-Niermeijer et al. (2000) and Canton and Slooff (1977) but in a design that was less suitable for standardisation e.g. because egg masses were used. But nevertheless, working with isolated eggs offers various advantages.
The development of a chronic mollusc toxicity test is a current work item on the agenda of the OECD. The freshwater pond snail Lymnaea stagnalis is one of the candidate snail species for such a test. This paper presents a 21-day chronic toxicity test with L. stagnalis, focussing on embryonic development. Eggs were collected from freshly laid egg masses and exposed individually until hatching. The endpoints were hatching success and mean hatching time. Tributyltin (TBT), added as TBT-chloride, was chosen as model substance. The selected exposure concentrations ranged from 0.03 to 10 μg TBT/L (all as nominal values) and induced the full range of responses. The embryos were sensitive to TBT (the NOEC for mean hatching time was 0.03 μg TBT/L and the NOEC for hatching success was 0.1 μg TBT/L). In addition, data on maximum limit concentrations of seven common solvents, recommended in OECD aquatic toxicity testing guidelines, are presented. Among the results, further findings as average embryonic growth and mean hatching time of control groups are provided. In conclusion, the test presented here could easily be standardised and is considered useful as a potential trigger to judge if further studies, e.g. a (partial) life‐cycle study with molluscs, should be conducted.
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^*: Address: F.A.C. Wiegant, Department of Molecular Cell Biology, Utrecht University, PO Box 80.056, 3508 TB Utrecht, The Netherlands Phone: 0031-30-2533972/3184; Fax: 0031-30-2513655

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Phyto-adaptogens protect against environmental stress-induced death of embryos from the freshwater snail Lymnaea stagnalis

Summary

J. Therm. Biol.

Toxicology

Pharmacol.

Ecotoxicol. Environm. Safety

Phytomedicine

Phytomedicine

J. Biol. Chem.

Phytomedicine

The role of inducible stress proteins in cardioprotective effects of Rhodiola rosea

J. Biokhimiya (Moscow)

Heat shock effects in snail development

Results and problems in cell differentiation

Evidence for two states of thermotolerance

Int. J. Hyperthermia

Heat-induced changes in thermosensitivity and gene expression during development

Dev. Growth Differ.

Thermotolerance induced by heat and ethanol

Int. J. Hyperthermia

Man and biologically active substance; the effect of drugs, diet and pollution on health