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THE EFFECT OF BIOLOGICALLY ACTIVE SUBSTANCES OF ARALIA MANDSHURICA AND DEUTERIUM DEPLETED WATER ON THE STRUCTURE OF SPLEEN IN MUS MUSCULLUS Cornelia Editoiu, Carmen Popescu, Greta Ispas, G. C. Corneanu , M. Zagnat , I. Stefanescu
VICTOR BABES CLINIC HOSPITAL OF INFECTIOUS DISEASES AND PNEUMOPHTISIOLOGY, CRAIOVA; EMERGENCY HOSPITAL, CRAIOVA; MEDICAL CLINIC, CRAIOVA; VASILE GOLDIS WEST UNIVERSITY, ARAD; GRIGORE TRAIAN POPA UNIVERSITY OF MEDICINE AND PHARMACY, IASI; N.R.-D.I. FOR CRYOGENICS AND ISOTOPES SEPARATION, RAMNICU VALCEA
Summary
In this experiment it was analyzed the effect of some biologically active substances extracted from the cortex of the Aralia mandshurica plants (Chisinau population) on laboratory mice (Mus muscullus), compared with a stress factor (X- ray radiation). The dilutions were made in distilled water and in deuterium depleted water. The combined administration of Aralia mandshurica plants cortex extracts presented a protective effect against X-ray radiation but also an immunostimulating effect, especially if the dilution was carried out with DDW. Key words: Aralia mandshurica, volatil oil, polyphenols, Mus muscullus, radioprotective effect
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Introduction
Aralia mandshurica (Araliaceae Fam., Dicotyledonatae Class, Angiospermatophyta Phylum) (Srbu, 1999) is a shrub of 3 to 6 meters high, originating from Primorie region in Siberia (Russia), grows spontaneously in the Far East and is inserted in culture, including in Moldavia. As a vegetable product, the plant roots are used because of the therapeutic properties (immunostimulating - Corneanu et al, 2003, stimulating and repairing Zagnat, 2004). The plant root contains triterpenoid saponoside - aralozide (4.7%), volatil oil (0.12%), flavonozide, resins and the aralin alkaloid (Nistreanu, 2000; Stanescu et al, 2004). In the Oriental traditional medicine are used extracts from the roots of species particularly in cases of physical and mental strain, convalescence, asthenic states, the beneficial effects being attributed to aralozides which have the property to increase the ability of adaptability of the organism in conditions of overload (Stanescu et al, 2002). 212 The deuterium depleted water / ultra light water (DDW) is low-deuterium water, 30-120 ppm deuterium; its physical aspect looks like colorless liquid, flavored water, with a low content in ion and hydrogendeuterium isotope. The effects of this kind of water are great: increased vascular reactivity, nonspecific immune stimulation, increased animal resistance to sublethal and lethal doses of radiation, remission of tumor cell lines, etc. (Titescu, 1999). DDW affects both the normal cells proliferation and the pathological ones, being involved in the processes of apoptosis and mechanisms anti carcinogens (Corneanu et al, 2005). In this experiment we analyzed the effect of administration of some biologically active substances in dilutions made with normal water or ultra light water, to the action of a stress factor, the X-rays. The experiment is part of a doctoral thesis done under the direction of Professor Doctor Corneanu Gabriel.
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Table 1. Mus muscullus experimental variants treated with bioactive substances diluted with distilled water
Treatment bioactive substances diluted with distilled water Untreated Volatil oil A. mandshurica Polyphenols A. mandshurica
Table 2. Mus muscullus experimental variants treated with bioactive substances diluted with deuterium depleted water
Variations Treatment bioactive substances diluted with deuterium depleted water non- irradiated C-DDW (control) Untreated V2 Volatil oil A. mandshurica V4 Polyphenols A. mandshurica
Spleen is the largest lymphoid organ of the body, with an important role in immune function and serves as a filter for the abnormal or senescent cells. The White Pulp is the spleen lymphoid component and includes B and T lymphocytes compartments. In terms of histology it consists of: periartehale pehlimfatice sheaths and splenic Malpighi corpuscles. The Periarteholare lymphatic sheaths (PALS) represent lymphoid tissue sleeves (timodependent areas) which contain 213
effector and regulatory T lymphocytes, interdigitating dendritic cells and macrophages. Red Pulp surrounds the white pulp and consists of splenic tissue cords (the Bilroth cords) situated among the venous sinuses. The Bilroth cords are composed of a three-dimensional network of reticular cells and reticular fibers, forming a functional unit with sinuses adventice. In this network, free spleen cells, with phagocytic function, move (macrophages). The core of these
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macrophages also shows nuclear appendix (Corneanu et al, 2007). At the Bilroth Cords level, the traffic has an open nature, because blood comes in direct contact with the spleen cells (Cesta, 2006, Raica et al, 2004). The process of apoptosis
plays an important role in the development of spleen, especially the lymphoid compartment, predominantly in the germ centers of B cells (Corneanu et al, 2005).
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Conclusions
X-radiations induce structural changes - Malpighi lymphatic follicle hypertrophy and hyperplasia, many lymphocytes present also in the red pulp and thin fibrous connective septa. The administration of DDW, only, produces structural alterations in the white pulp, with the presence, in the red pulp of lymphocytes whose cell membrane is broken; the lymphocyte alteration induced by DDW being stronger than the one induced by X-rays. The administration of essential oil from the cortex of Aralia mandshurica, on non-irradiated variants produces reduced inflammatory infiltrates, probably due to the monoterpenes hydrocarbons contained in the essential oil which, diluted in DDW, has an antiseptic, anti-inflammatory and immunomodulating effect. Similar issues were reported in other studies, too (Istudor, 2001). Acknowledgements These researches are part of the grant 33.062/2004, sponsored by CNCSISBucharest. The authors address sincere thanks to Mr. Dr. Marin Popa, head of Flaw Laboratory from SC Electroputere SA In irradiated variants, administration of the essential oil diluted with DDW shows a stronger protective effect on spleen structure than in distilled water, observed in splenic follicles hyperplasia, red pulp expansion and immunostimulative by increasing the number of megakaryocytes. In case of using the polyphenols extract from the cortex of Aralia mandshurica diluted with DDW has a protective effect on the structure of spleen, observed in the marked hypertrophy of the red pulp by intense dilation of capillary sinusoids and with the presence of numerous megakaryocytes. The flavones derivatives develop antiinflammatory, anti-allergic, antimicrobials, antioxidant and capillary protective effects (Stanescu et al, 2002).
Craiova, who performed X-irradiation of animals. Also thank Dr. Mr. Marin Zagnat, UMF Iasi, which used to extract bioactive substances (volatile oil and polyphenols) from the cortex of Aralia mandshurica.
References
Cesta MF., 2006, Normal structure, function, and histology of the spleen. Toxicol. Pathol. 34:455465. Corneanu G., Craciun C., Ciupina V., Prodan G., Corneanu M., Atyim P., Stefanescu I., Iacob M., Corneanu L-M., 2005, The efect of Nigella sativa L. alkaloids and of deuterium-depleted water on Mus musculus L. spleen, Revue de Cytologie et Biologie vegetales-Le Botaniste. 28 (s.i.). 84-91. Corneanu G., Corneanu M., Atyim P., Craciun C., 2007, The COX-2 gene implication in the immune response (experiments on animals, Mus musculus L.), Analele Universitatii din Craiova, Agricultura-Montanologie-Cadastru. vol. XXXVII/A. 525-535. Istudor V., 2001, Farmacognozie. Fitochimie si fitoterapie, Ed. Medicala, Bucuresti. Nistreanu A., 2000, Farmacognozie. USMF ..Nicolae Testemitanu", Chisinau. Raica M., Mederle O., 2004, Histologie teoretica si practica, Ed. Brumar, Timisoara: 287-301, 137140. Sarbu A., 1999, Biologie vegetala, Note de curs, Editura Universitatii Bucuresti. Stanescu U., Miron A., Hancianu M., Aprotosoaie C., 2002, Bazele farmaceutice, farmacologice si clinice ale fitoterapiei, (vol. II). Ed. Gr. T. Popa , Iasi, 76-145. Stanescu U., Miron A., Hancianu M., Aprostosoaie C., Plante medicinale de la A la Z, monografii ale plantelor de interes terapeutic, 2004, (vol. II). Ed. Gr. T. Popa , UMF Iasi, 613. 10. Titescu G., Iliescu R., Nastasa V., Bild W., Stefanescu I., Haulica I., Lupusoru C., 1999, Apa echilibrata in deuteriu. Research concerning the radioprotective and immunostimulating efects of deuterium-depleted water, Rom. J. Physiol. 36 (3-4): 18-205.
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Figura 2. Thin fibro conjunctive septa of mouse spleen (oc. 10 x ob. 20) C-X
Figura 3. Numerous lymphocytes at the red pulp level (oc. 10 x ob. 10) C-DDW
Figura 4. Big splenic follicles of mouse spleen (oc. 10 x ob. 10) C-DDW-X
Figura 5. Hyperplasia red pulp level and rare megakaryocytes of mouse spleen (oc. 10 x ob. 10) V2
Figura 6. Dilatation of red pulp capillary and hypertrophy of fibro conjunctive septa of mouse spleen (oc. 10 x ob. 40) V2-X
Figura 7. Numerous follicles with germ centers of mouse spleen (oc. 10 x ob. 20) V4
Figura 8. Atrophy of white pulp of mouse spleen (oc. 10 x ob. 10) V4-X
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