In vitro effect of short peptides on expression of interleukin-2 gene in splenocytes

Synthetic peptides Vilon (Lys-Glu), Epithalon (Ala-Glu-Asp-Gly), and Cortagen (Ala-Glu-Asp-Pro) in vitro activated interleukin-2 mRNA synthesis in splenocytes from CBA mice in the absence of specific inductors. The intensity of interleukin-2 mRNA synthesis in splenocytes depended on the type, concentration, and duration of treatment with the peptides. Vilon and Epithalon were most potent, while Cortagen produced a less pronounced effect on interleukin-2 mRNA synthesis.     

Tissue-Specific Effects of Peptides

Synthetic peptides (cytogens) Cortagen, Epithalon, Livagen, and Vilon stimulated the growth of explants from rat brain cortex, subcortical structures, liver, and thymus, respectively, in organotypic cultures. These peptides produced tissue-specific effects: they stimulated the growth of explants from tissues, whose cytomedins (peptide complexes) were used for chemical synthesis.     

Effects of Short Peptides on Lymphocyte Chromatin in Senile Subjects

Effects of synthetic short peptides (Vilon, Epithalon, Livagen, Prostamax, and Cortagen) on activity of ribosome genes, parameters of common heterochromatin melting, polymorphism of structural heterochromatin (C segments) of chromosomes 1, 9, and 16, and variability of facultative heterochromatin were studied in leukocytes of subjects aged 75-88 years. All the studied peptides induced activation of ribosome genes, decondensation of densely packed chromatin fibrils, and release of genes repressed as a result of age-specific condensation of the cellular euchromatin regions (deheterochromatinization of facultative chromatin). Treatment with Epithalon, Livagen, and Prostamax led to decondensation of chromosome 1 pericentromeric structural chromatin, while Epithalon and Livagen treatment led to changes in chromosome 9 as well. Hence, short peptides activate heterochromatin and heterochromatinized regions of cell chromosomes in senile subjects.     

Studies of the effects of Vilon and Epithalon on gene expression in mouse heart using DNA-microarray technology

Expression of 15,247 clones from a cDNA library in the heart of mice receiving Vilon and Epithalon was studied by DNA-microarray technology. We revealed 300 clones (1.94% of the total count), whose expression changed more than by 2 times. Vilon changed expression of 36 clones, while Epithalon modulated expression of 98 clones. Combined treatment with Vilon and Epithalon changed expression of 144 clones. Vilon alone or in combination with Epithalon activated expression of 157 clones (maximally by 6.13 times) and inhibited expression of 23 clones (maximally by 2.79 times). Epithalon alone or in combination with Vilon activated expression of 194 clones (maximally by 6.61 times) and inhibited expression of 48 clones (maximally by 2.71 times). Our results demonstrate the specific effects of Epithalon and Vilon on gene expression.     

Expression of Argyrophilic Proteins in the Nucleolar Organizer Regions of Human Thymocytes and Thymic Epitheliocytes under Conditions of Coculturing with Vilon and Epithalon Peptides

Vilon stimulated and Epithalon suppressed the expression of argyrophilic proteins in nucleolar organizer regions of thymocytes and epithelial cells, stimulating or reducing, respectively, the formation, assembly, and transport of ribosomes into the cytoplasm and thus determining the intensity of protein synthesis in these cells. A direct mitogenic effect of Vilon was also revealed: this peptide promoted thymocyte transformation into proliferating blast cells.     

Production of lymphocyte-activating factors by mouse macrophages during aging and under the effect of short peptides

Age-specific characteristics of production of lymphocyte-activating factor by mouse peritoneal macrophages and modulation of this production by short synthetic peptides (Vilon, Epithalon, and Cortagen) were studied. The production of lymphocyte-activating factors by macrophages stimulated with lipopolysaccharides in vitro was lower in old animals. The opposite modulating effects of short peptides on the production of lymphocyte-activating factors by resident and lipopolysaccharide-stimulated macrophages in young and old mice were demonstrated for the first time. This is a possible mechanism of immune system dysfunction during aging, which opens new vistas for its correction with short synthetic peptides. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 142, No. 9, pp. 333–336, September, 2006     

Effect of Vilon and Epithalon on glucose and glycine absorption in various regions of small intestine in aged rats

Vilon (Lys-Glu) and Epithalon (Ala-Glu-Asp-Gly) administered orally for 1 month improved transport characteristics of the small intestine in aged rats. Vilon enhanced passive glucose accumulation in the serous fluid in inverted sac made from the distal region of the small intestine, while Epithalon enhanced this process in the medial region. Vilon stimulated active glucose accumulation in the serous sac of the medial small intestine, Epithalon - in the proximal and distal small intestinal segments. Glycine absorption increased only in the proximal intestinal segment under the effect of Epithalon.     

Synthesis of IL-2 mRNA in Cells of Rat Hypothalamic Structures after Injection of Short Peptides

In situ hybridization on paraffin sections of the rat brain showed that synthetic peptides Vilon, Epithalon, and Cortagen modulated the expression of IL-2 gene in vivo in cells of some hypothalamic structures depending on the terms and routes of administration.__________Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 139, No. 6, pp. 688–690, June, 2005     

Effects of Peptides on Generation of Reactive Oxygen Species in Subcellular Fractions of Drosophila Melanogaster

We studied the effects of Epithalon (Ala-Glu-Asp-Gly) and Vilon (Lys-Glu) on free radical processes in highly inbred HA⁺line of Drosophila melanogaster. Vilon inhibited generation of reactive oxygen species in mitochondria, but stimulated this process in the cytosol. We found sex- and age-related differences in the generation of reactive oxygen species and cytosol antioxidant activity.     

Epithalon inhibits tumor growth and expression of HER-2/neu oncogene in breast tumors in transgenic mice characterized by accelerated aging

Female transgenic FVB mice carrying breast cancer gene HER-2/neu were monthly injected with Vilon or Epithalon (1 g subcutaneously for 5 consecutive days) starting from the 2nd month of life. Epithalon markedly inhibited neoplasm development: the maximum size of breast adenocarcinomas was 33% lower than in the control (p<0.05). The intensity of HER-2/neu mRNA expression in breast tumors of Epithalon-treated mice was 3.7 times lower than in control animals. These results indicate that Epithalon inhibits breast tumor development in transgenic mice, which is probably related to suppression of HER-2/neu expression.     

Interleukin-2 concentration in hypothalamic structures of rats receiving peptides during mild stress

The number of hypothalamic IL-2-containing cells changed in rats receiving Vilon and Epithalon during mild stress (handling). The number of IL-2-positive cells in hypothalamic structures decreased 24 h after intramuscular injection of Epithalon and 2 h after intranasal administration of the test peptides. Adaptation of animals to experimental conditions prevented the decrease in the number of IL-2-positive cells in the supraoptic nucleus after intranasal administration of Epithalon. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 141, No. 4, pp. 371–374, April, 2006     

Enhancement of in vitro lipopolysaccharide-stimulated interleukin-1 production by levamisole

The production of interleukin-1 (IL-1) by the P388D1 mouse macrophage cell line and by adherent peritoneal exudate cells (PMs) was examined. In vitro IL-1 production by P388D1 cells treated with lipopolysaccharide (LPS) was enhanced by coculture with levamisole (0.1 to 10 microM). Oral administration of levamisole (3 mg/kg) to mice also resulted in potentiation of in vitro IL-1 production by thioglycollate-elicited peritoneal macrophages in response to in vitro LPS stimulation. Potentiation was approximately twofold. IL-1 production in the absence of LPS by either the P388D1 cells or the PMs was nil, and levamisole did not directly stimulate IL-1 production in these cases. IL-1 activity in the culture supernatants was measured by thymocyte comitogenic assays. The immunochemical identify of the thymocyte comitogenic activity as IL-1 alpha was confirmed by neutralization with a specific goat anti-mouse IL-1 alpha antiserum. These results suggest that one mechanism by which levamisole acts to normalize and restore immune responses may be enhancing the signals which enable activated macrophages to secrete IL-1.     

Effect of pineal tetrapeptide on antioxidant defense inDrosophila melanogaster

Effect of synthetic pineal tetrapeptide L-Ala-L-Glu-L-Asp-L-Glu (Epithalon) on specific catalase activity and the content of conjugated hydroperoxides in highly inbredDrosophila melanogaster lines differing in reproductive functions were studied. It was shown that Epithalon is a potent modulator of the antioxidant defense, whose biological activity 1000-fold surpasses that of the complex pineal peptide preparation Epithalamin.     

A macrophage-derived factor induced by alpha 1-acid glycoprotein that inhibits IL-1 comitogenic activity

After exposure to a concanavalin A (Con A)-unreactive variant of alpha 1-acid glycoprotein (AGP), macrophages released an inhibitor of interleukin-1 (IL-1) proliferative activity in the thymocyte comitogenic assay. This effect was observed with AGP concentrations above 100 micrograms/ml in the macrophage supernatant and would appear to be mediated by the macrophages, since native AGP had no activity on thymocyte proliferation. Preliminary physicochemical characterization showed that the factor was partially resistant to heating, undialyzable, and eluted with an apparent molecular mass of 50-100 kDa when subjected to Sephacryl S-200 chromatography. Murine IL-1 and human (h) recombinant (r) IL-1 were affected by this factor to the same extent. IL-1 and IL-2 co-induced thymocyte proliferation, which is mitogen-independent, was also inhibited, whereas hrIL-2 activity was not suppressed when assayed in thymocytes with PHA at a submitogenic concentration or in CTLL cells. The factor did not interfere with TNF alpha or hrIL-6 activity when tested against their specific cell line. These data indicate that the inhibitor may act specifically against IL-1 activity and further elucidate the possible role of AGP in the modulation of IL-1 activity via the secretion of an inhibitor.     

Effects of peptides on proliferative activity of retinal and pigmented epithelial cells

We studied the effects of Retinalamin (polypeptide preparation isolated from the retina) and a synthetic peptide Epithalon (Ala-Glu-Asp-Gly) on proliferative activity of retinal and pigmented epithelial cells. Experiments showed that Retinalamin and Epithalon (in certain concentrations) tissue-specifically stimulated proliferation of retinal and pigmented epithelial cell in culture.     

Epithalon decelerates aging and suppresses development of breast adenocarcinomas in transgenic her-2/neu mice

Female transgenic FVB/N mice carrying the breast cancer gene HER-2/neu received epithalon (Ala-Glu-Asp-Gly) in a dose of 1 mg subcutaneously 5 times a week to from the 2nd month of life to death. Epithalon prolonged the average and maximum lifetimes of mice by 13.5 (p<0.05) and 13.9%, respectively. The peptide prolonged the average lifetime of animals without neoplasms (by 34.2%, p<0.05). Epithalon decelerated the development of age-related disturbances in reproductive activity and suppressed the formation of neoplasms. The peptide decreased the incidence of breast adenocarcinomas, lungs metastases (by 1.6 times, p<0.05), and multiple tumors (by 2 times). Epithalon 3.7-fold increased the number of mice without breast tumors (p<0.05), while the number of animals with 6 or more breast tumors decreased by 3 times (p<0.05). Epithalon prolonged the lifetime of mice with breast tumors by 1.4 times (p<0.05). These results indicate that Epithalon possesses geroprotective activity and inhibits breast carcinogenesis in transgenic mice, which is probably related to suppression of HER-2/neu expression.     

Thymic reticulum in mice III. Phagocytic cells of the thymic reticulum in culture secrete both prostaglandin E2 and interleukin 1 which regulate thymocyte proliferation

A phagocytic cell of the thymic reticulum in culture (P-TR-C) has recently been isolated whose characteristics closely resemble those of the thymic interdigitating cell despite the modifications induced by culture conditions. In the present report it is demonstrated that this cell population is able to produce both interleukin 1 (IL1) and prostaglandin E₂ (PGE₂). IL1 has no direct mitogenic effect on thymocytes but is comitogenic with concanavalin A. PGE₂ has no direct effect on the secretion of IL1 by P-TR-C, but has an effect antagonistic to IL1, inhibiting thymocyte proliferation. P-TR-C are thus able to modulate thymocyte proliferation in vitro.     

Effect of the dipeptide vilon on activity of digestive enzyme in rats of various ages

Peroral administration of Vilon (Lys-Glu) to male and female Wistar rats aging 3 and 11 months changed activity of digestive enzymes (invertase, maltase, alkaline phosphatase, and amino- and dipeptidases) in various portions of the gastrointestinal tract. The increase in enzyme activity was most pronounced in 11-month-old animals, which diminished differences between rats of various ages. Our results indicate that Vilon produces positive effects on digestive enzyme activity during aging.     

苯二氮类药物对细胞免疫功能的影响

苯二氮类药物（安定）能促进小鼠胸腺细胞增殖,这种作用以安定5mg/kg腹腔注射,每天一次连续给药5d的作用最强。安定对小鼠脾脏细胞增殖、自然杀伤（NK）细胞毒活性、脾细胞初次抗体产生以及对体外培养胸腺细胞增殖等均无明显影响。安定对截肢应激所致小鼠胸腺、脾脏细胞免疫抑制有拮抗作用,并可有效地拮抗应激导致的小鼠胸腺重量减轻。