Modeling of Benign Prostatic Hyperplasia in Rats with a High Dose of Testosterone.

In order to optimize the testosterone model of benign prostatic hyperplasia, we studied the effect of castration and different doses of testosterone on the induction of the proliferative process in the prostate of Wistar rats. It was shown that 4-week subcutaneous administration of testosterone propionate in a dose of 20 mg/kg causes pronounced proliferative and hemodynamic disorders in the dorsolateral gland morphologically similar in castrated and non-castrated males. Administration of testosterone in a dose of 3 mg/kg had no significant effect on the dynamics of the pathological process in non-operated rats and normalized the structure of the gland in castrated animals. Morphological study showed that castration of males provides no visible advantages in reproducing the testosterone model of benign prostatic hyperplasia. The proposed non-traumatic modification of the model with a high dose of testosterone has good reproducibility and sensitivity to therapeutic agents, as shown by the example of finasteride.

Prostatotropic Action of Glycyrrhizic Acid Disodium Salt in Benign Prostatic Hyperplasia Models.

The prostatotropic activity of glycyrrhizic acid disodium salt (Na2GA) was studied in the models of benign prostatic hyperplasia (BPH) induced by chronic injection of sulpiride (40 mg/kg intraperitoneally for 8 weeks) or testosterone propionate (20 mg/kg subcutaneously for 4 weeks) in the Wistar rats. The oral administration of Na2GA in a dose of 100 mg/kg produced a moderate antiproliferative effect in both BPH models resulting in reduced volume density of prostatic epithelium (in the testosterone model) and increased volume density of the glandular lumen (in both models). The observed prostatotropic effects of Na2GA were similar to those of Permixon and finasteride, but they were less pronounced as confirmed by smaller drops in epithelial volume density and epithelial-to-stromal ratio compared to the effects of both reference drugs.

Ultrastructure of the Liver in Response to Cyclophosphamide and Triterpenoids.

Ultrastructural reorganization of liver cells after isolated injections of cyclophosphamide, betulonic acid or its ß-alanylamide, and combined treatment with the cytostatic and each of the triterpenoids is studied. Cyclophosphamide causes significant ultrastructural changes in all intracellular compartments of hepatocytes. Both triterpenoids cause moderate cytotoxic and stimulatory effects on the liver cell populations (hepatocytes, sinusoidal endotheliocytes, and Kupffer cells), when used alone. The cytotoxic effect of betulonic acid manifests in modification of the fine structure of hepatocyte mitochondria, sequestration of glycogen, intensification of autophagic processes, emergence of necrobiotic changes in hepatocytes and endotheliocytes; betulonic acid amide actively modifies the mitochondrial fine structure (hypertrophic organelles, matrix rarefaction, uneven dilatation of cristae). The effects of combinations of cyclophosphamide with betulonic acid or its amide on liver are polytarget: the cytotoxic activity of the cytostatic is potentiated towards some cells, while in other cells the regeneratory reactions are stimulated. The common cytological cytoprotective effects of betulonic acid and its amide used alone and in combination with cytostatics include stimulation of the endocytotic (pinocytotic) activities of the cells and stimulation of intracellular regeneration processes in them.

Morphological Assessment of Prostatotropic Activity of (3,5-Dimethyl-4-Hydroxy)Benzyl Thiododecane on a Model of Benign Prostatic Hyperplasia in Rats.

Prostatotropic activity of (3,5-dimethyl-4-hydroxy)benzyl thiododecane (T-DD) was evaluated on a model of benign prostatic hyperplasia induced in Wistar rats by chronic (2 months) intraperitoneal administration of sulpiride (40 mg/kg). Morphometric analysis of the dorsolateral lobe of the prostate showed that after the 2-month course of intragastric T-DD (100 mg/kg) administered in parallel with sulpiride, the volume density of glandular epithelium decreased by 1.7 times, while the volume density of prostate stroma increased by 2 times. After administration of the reference drug Permixon at a dose of 50 mg/kg, the volume densities of epithelium decreased by 1.3 times and stromal volume density increased by 1.5 times. The observed effects are presumably related to suppression of 5α-reductase activity and modulation of estrogen receptors in the prostate.

Cardiotoxic and Dyslipidemic Effects of Doxorubicin and Betulinic Acid Amide.

Changes in the blood lipid spectrum and structural reorganization of the rat myocardium in response to injection of a single sublethal dose of doxorubicin (7 mg/kg) alone and in combination with course administration of betulonic acid amide (100 mg/kg/day for 14 days) were studied. Betulinic acid amide in the specified dose exhibited less pronounced cardiotoxic (necrobiotic impairment of cardiomyocytes) and dyslipidemic (increase of cholesterol and triglyceride levels) effects in comparison with doxorubicin. Combined treatment with betulinic acid amide and doxorubicin led to more pronounced remodeling of the myocardium, which was shown by a significant increase of the connective tissue/cardiomyocyte volume ratio detected by day 14 of the experiment.

Specific Features of Progression of the Parasitic Invasion, caused by Opisthorchis felineus, in Golden Hamsters.

The influence of Opisthorchis felineus invasion on the development of pathological changes in the hepatobiliary system was studied in 120 golden hamsters in a long-term experiment (42 weeks) after single infection per os in the dose of 50 metacercariae per animal. The animals were sacrificed on weeks 4, 8, 12, 16, 28 and 42. Chronic experimental infestation with O. felineus triggered a cascade of morphogenetic processes in both extrahepatic and intrahepatic biliary systems. At the early stages of the experiment, polyps and strictures of bile ducts were formed in the lobar bile ducts; in portal tracts, hyperplasia and adenomatous transformation of the newly formed epithelial structures were observed. At the later stages, third-degree biliary intraepithelial neoplasia developed in the lobar bile ducts; in the intrahepatic bile ducts, increased epitheliocyte hyperplasia and invasive growth of cell cords were observed, that impaired tissue architectonics. Progressing cell atypia can be classified as cholangiocellular cancer.

[Synthesis and Anti-Inflammatory activity of the propargylamino derivatives of naphthoquinonlevopimaric acid].

Modification of naphthoquinonlevopimaric acid was carried out by introducing of propargylamino residues. Anti-inflammatory activity Mannich bases were studied.

[Synthesis of new analogues of combretastatin A-4 and the study of their anti-inflammatory activity].

New approach to synthesis of analogs of natural Combretastatin A-4 based on interaction of α-acetylenic ketones with secondary amines (diethyl amine, pyrrolidine, piperidine, morpholine) is offered. Unknown analogs of Combretastatin A-4 with ß-aminovinylcarbonyl bridges are received earlier. Anti-inflammatory activity of the received connections is studied.

Hepatoprotective properties of betulonic acid amide and heptral in toxic liver injury induced by carbon tetrachloride in combination with ethanol.

Toxic liver injury with the development of fibrosis and cirrhosis was modeled in Wistar rats by intragastric administration of 0.1 ml/kg CCl4 in combination with 5% ethanol with glucose 3 times a week for 6 weeks. The animals were treated with betulonic acid amide (50 mg/kg in Tween aqueous solution) and heptral (6 mg/kg) as hepatoprotective compounds. It was found that betulonic acid amide stimulated the regenerative response in hepatocytes under conditions of combined toxic exposure and promoted recovery of their qualitative and quantitative characteristics, which was accompanied by a significant decrease in the severity of liver fibrosis and the absence of cirrhotic transformation of the liver.

Complex investigation of the effects of lambertianic acid amide in female mice under conditions of social discomfort.

The effects of chronic administration of a new substance lambertianic acid amide and previously synthesized methyl ester of this acid were compared in female mice living under conditions of social discomfort. For modeling social discomfort, female mouse was housed for 30 days in a cage with aggressive male mouse kept behind a transparent perforated partition and observed its confrontations with another male mouse daily placed to the cage. The new agent more effectively than lambertianic acid methyl ester improved communicativeness and motor activity of animals, reduced hypertrophy of the adrenal glands, and enhanced catalase activity in the blood. These changes suggest that lambertianic acid amide produces a pronounced stress-protective effect under conditions of social discomfort.

Soluble chitosan-carrageenan polyelectrolyte complexes and their gastroprotective activity.

The soluble polyelectrolyte complexes (PEC) κ-carrageenan (κ-CG):chitosan was obtained. Binding constant value (2.11 × 10(7)mol(-1)) showed high affinity of κ-CG to chitosan. The complex formation of κ-CG:chitosan 1:10 and 10:1 w/w was shown by centrifugation in a Percoll gradient. Using atomic force microscopy we showed that the supramolecular structure of the complexes is different from each other and from the macromolecular structure of the initial polysaccharides. The gastroprotective and anti-ulcerogenic effect of κ-CG, chitosan and their complexes was investigated on the model of stomach ulcers induced by indometacin in rats. PEC κ-CG:chitosan have gastroprotective properties which depend on their composition. Complex κ-CG:chitosan 1:10 w/w possesses higher gastroprotective activity than the complex 10:1 w/w. These results suggest that the gastroprotective effect of complexes can be associated with their protective layer on the surface of the mucous membrane of a stomach, which avoids a direct contact with the ulcerogenic agent.

Expression of chicken CTCF gene in COS-1 cells and partial purification of CTCF protein.

The chicken gene for transcription factor CTCF was expressed in COS-1 mammalian cells. The CTCF protein containing polyhistidine tag was partially purified using metallo-affinity and ion-exchange chromatography. The expressed protein localized in the cell nucleus and was shown to be functionally active in the electrophoretic mobility shift assay and specifically interacted with anti-CTCF antibodies.

[Antitumor activity of dihydrobetulonic acid amides in vitro and in vivo].

Amides with homopiperidinic and piperazinic cycles were synthesized from dihydrobetulonic acid which was obtained by dihydrobetulin oxidation. All substances have shown high antitumor activity (CCID50 3.5-36.2 microM) in vitro in lymphoid (CEM-13, U-937) and monocytic (MT-4) human cell lines. Amides with methyl- and ethyl-piperazinic residues don't influence viability of Lung Lewis Carcinoma cell in culture and haven't any significant effect to its transplantates in C57BL/6 mice. But such amides inhibit efficiently the metastatic elaboration in lung of these mice. The antimetastatic activity increases followed by the change of aliphatic residue length in piperazinic cycle from methyl to ethyl.

Correction of drug-induced hepatotoxicity with new triterpene derivatives in transplanted RLS lymphoma.

The hepatoprotective effects of new triterpene derivatives, betulin 3ß,28-di-O-nicotinate (of3) and 3,20-dioximino-29-norlup-28-ic acid methyl ester (of15), were studied in CBA/Lac mice with transplanted RLS lymphoma receiving polychemotherapy and without it. Injection of of3 and of15 agents to animals with tumors receiving polychemotherapy reduced the severity of toxic involvement of the liver, reduced mitotic activity of tumor cells in the primary node in animals receiving and not polychemotherapy, and produced a moderate antitumor effect. These effects were more pronounced for of15 agent. In addition, injection of agents of3 and of5 to animals with transplanted RLS lymphoma reduced the intensity of alterations associated with the total systems and local effects of the neoplastic process.

[Hepatoprotective activity of betulonic acid amides containing a piperidine or pyrrolidine nitroxide moiety].

Betulonic acid amides containing a nitroxyl radical moiety possess antiholestatic effects in mice. Introduction ofpiperidine nitroxide moiety into lupan core increases its hepatoprotective activity. Oral administration of piperidine nitroxide derivative in dose 50 mg/kg doesn't stimulate transplanted tumor growth and raises a lifespan of mice.

[Synthesis of aminopropylaminoderivatives of betulinic and oleanolic acids].

On the basis of betulinic and oleanolic acids triterpenoids with different with different amine fragments: (3-aminopropoxy)-, 3-acetyl-(3-aminopropyl)amino-, 6-[bis(3-aminopropyl)amino]hexylamino-, (3-aminopropyl)-4-aminosulfonyl-4-phenylamino- at positions C3 and C28 were synthesized. It is shown that betulonic acid amide with 4,4'-diaminodiphenylsulfonic substituent don't render antitumor effect in mice with transplantable Lewis lung carcinoma, but possess significant anti-inflammatory activity.

Effect of triterpene derivatives on the total hepatocyte count in the liver of rats with toxic hepatitis.

We studied hepatoprotective activity of betulonic acid and its alaninamide on the model of combined CCl(4)- and ethanol-induced toxic liver damage in rats. The test substances, especially betulonic acid alaninamide, considerably reduced the elevated biochemical parameters in animals with toxic liver damage. Betulonic acid alaninamide also stimulated reparative processes in the liver (activated hepatocyte proliferation). Heptral (reference drug) produced no appreciable effects on the reparative processes. Our findings suggest that betulin derivatives exhibit pronounced protective properties.

Study of skin anti-ageing and anti-inflammatory effects of dihydroquercetin, natural triterpenoids, and their synthetic derivatives.

Accessible triterpenoids of ursane and lupane series, the flavonoid dihydroquercetin and their synthetic derivatives with polar substituentss were tested in vitro for inhibition of collagenase 1 (MMP-1) in UVB irradiation assay. Ursolic acid and uvaol disuccinate were the most active inhibitors in the ursane series. In the lupane series, the best inhibition was manifested by carboxymethyl ester of betulonic acid and betulin succinates. Down- regulation of MMP-1 by dihydroquercetin and its synthetic derivatives surpassed the activity of a standard (retinoic acid).

Structure of the liver in mice with transplanted Lewis carcinoma during polychemotherapy and correction with betulonic acid and its derivatives.

Morphological and morphometric studies of the liver with transplanted Lewis carcinoma were performed after polychemotherapy and correction with betulonic and [3-oxo-20(29)-lupene-28-oil]-3-aminopropionic acids and their methyl esters. It was demonstrated than betulonic and [3-oxo-20(29)-lupene-28-oil]-3-aminopropionic acids reduced the degree of degenerative changes and volume density of necrotic changes in hepatocytes after polychemotherapy. Methyl esters of these acids little changed the severity and spreading of destructive and necrotic changes in the liver caused by complex cytostatic therapy. It was also shown that all studied triterpenoids exhibited more pronounced antimetastatic effect (evaluated by the decrease in volume density of liver metastases) compared to polychemotherapy.