Modeling of Hypervolemia in Pulmonary Circulation in Rats Changes the Structure of NO-Mediated Relaxation of Pulmonary Arteries.

We analyzed the contribution of soluble guanylate cyclase-dependent pathway into NO-mediated relaxation of pulmonary arteries under conditions of high pulmonary blood flow modeled by creation of carotid artery-jugular vein shunt in rats. Inhibitor of soluble guanylate cyclase suppressed NO-donor induced relaxation was lower in rats with shunt, but dilatation in response to phosphodiesterase V inhibitor did not differ in the sham-operated and shunt groups. Thus, the structure of NO-mediated vasodilatation of pulmonary arteries under conditions of hypervolemia of pulmonary circulation was shifted to soluble guanylate cyclase-independent pathways, whereas intracellular soluble guanylate cyclase-dependent mechanisms of dilatation were in general unchanged.

The Structure of NO-Mediated Dilatation of Pulmonary Arteries Depends on NO Availability.

NO-mediated vasodilatation can be realized via two pathways: dependent and independent on soluble guanylate cyclase; the latter is implemented through NO interaction with ionic channels. We evaluated the contribution of these pathways into NO-induced relaxation of isolated pulmonary arteries in rats. In pulmonary arteries, in contrast to systemic vessels, soluble guanylate cyclase-independent mechanisms is more important, because it mediates relaxation in response to low concentrations of NO donor. The role of soluble guanylate cyclase-dependent mechanisms in the mechanisms of vasodilatation increases with increasing NO donor concentrations.

Inducible NO Synthase mRNA Expression and Infiltration of Rats Myocardium with Inflammatory Cells in 2-4 Hours after Modeled Permanent Ischemia or Ischemia/Reperfusion.

Expression of inducible NO-synthase mRNA and myocardial infiltration with neutrophils were studied in rats with modeled permanent ischemia and ischemia/reperfusion models. Expression of inducible NO synthase mRNA in the ischemic region increased significantly in 3, 3.5, and 4 h in modeled ischemia/reperfusion and in 3.5 and 4 h in permanent ischemia. Myocardial infiltration with neutrophils was significantly higher than in intact controls throughout the experiment without significant intergroup differences. In non-ischemic myocardium, enhanced expression of inducible NO synthase mRNA and moderate neutrophilic-lymphocytic myocardial infiltration were also observed in 3.5, and 4 h after ischemia.

[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.

Disorders in cytokine gene expression in endometrial hyperplasia and effect of hormone therapy.

We studied local expression of insulin-like growth factor 1, insulin-like growth factor receptor, epithelial growth factor, transforming growth factor beta2, PCNA, TNF-alpha, type I TNF receptor, Fas, FasL, IFN-gamma, IL-1beta, IL-4, IL-6, IL-8, IL-10, and IL-12 genes in intact and hyperplastic endometrium. Endometrial hyperplasia was associated with reduced production of TNF-alpha (p<0.05), PCNA (p<0.05), and epithelial growth factor mRNA and enhanced production of Fas mRNA (p<0.01). The expression of TNF-R1, IL-1beta, and IL-12 genes decreased only in glandular cystic hyperplasia (p<0.05 for all genes), expression of insulin-like growth factor 1 gene decreased only in adenomatous hyperplasia (p<0.05). Dufaston therapy of glandular cystic hyperplasia and zoladex therapy of adenomatous hyperplasia normalized expression of Fas receptor, PCNA, and insulin-like growth factor 1 genes, while the expression of IFN-gamma and IL-6 genes, which was normal in hyperplasia, decreased (p<0.05). Zoladex therapy decreased the production of transforming growth factor beta2 (p<0.05) and IL-1beta (p<0.01) mRNA, dufaston therapy decreased production of TNF-alpha (p<0.05) and IL-4 mRNA (p<0.05). Hence, both apoptosis and proliferative activity were suppressed in endometrial hyperplasia, and hormone therapy created prerequisites for transition of the endometrium into the normal proliferation stage.

Location and intensity of IL-8 and TGF-beta2 mRNA production in the fimbrial compartment of fallopian tubes and IL-10 in the endometrium in patients with pyoinflammatory adnexal diseases.

The location and level of IL-8 and TGF-beta2 expression in the fimbrial compartment of fallopian tubes and IL-10 expression in the endometrium of women with pyoinflammatory adnexal diseases were studied by in situ hybridization. These diseases are associated with considerable changes in the levels of local production of these cytokines. Inflammatory infiltration and epithelial cells were most active producers of IL-9 and TGF-beta2 in the fimbrial compartment of fallopian tubes, while in the endometrium IL-10 gene was expressed at a high level primarily in the glandular epithelial cells.

Apoptosis in fimbriae of fallopian tubes and endometrium in pyoinflammatory adnexal diseases.

In patients with pyoinflammatory adnexal diseases moderate or intense DNA degradation was observed in the majority of epithelial, stromal, and inflammatory infiltration cell in inflammatory foci in the fimbrial compartment of fallopian tubes without signs of tissue destruction. Expression of TNFR1 gene increased 2.7-fold and expression of Fas gene decreased 3.1-fold compared to intact endosalpinx, which indicates induction of apoptosis triggered by tumor necrosis factor-alpha and inhibition of the Fas-dependent pathway. No signs of apoptosis were detected in the endometrium. Generalized apoptosis in the fimbrial compartment of the tubes can be a mechanism limiting the inflammatory process.

Local expression of cytokine genes in uterine adnexa and endometrium of women with pyoinflammatory adnexal diseases.

Changes in the local expression of IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-15, IL-18, TNF-alpha, IFN-gamma- and TGF-beta(2) genes in the uterine adnexa and endometrium were studied in women with pyoinflammatory adnexal diseases. Examination of tissue specimens from the uterine adnexa involved in inflammation revealed a direct correlation in the levels of mRNA production between IL-6 and IL-10 (r=0.93, p<0.1), IL-6 and IL-4 (r=0.96, p<0.01), IL-10 and IL-4 (r=0.91, p<0.01), IL-12 and IFN-gamma (r=0.98, p<0.01). Expression of IL-4 gene increased 5.1-fold (p=0.001), IL-6 2-fold (p=0.007), IL-8 90.2-fold (p=0.009), IL-10 2.9-fold (p=0.008), IL-12 2.3-fold (p=0.3), and TGF-beta(2) gene 10.3-fold (p=0.1). In the endometrium of women with pyoinflammatory adnexal diseases only IL-10 gene expression increased (15.6-fold, p=0.007).

Correlations in the cytokine system in endometrial hyperplasia.

Correlations between local expression of insulin-like growth factor 1, insulin-like growth factor receptor, epithelial growth factor, transforming growth beta2 factor, PCNA, TNF-alpha, TNF receptor 1, Fas, FasL, IFN-gamma, IL-1beta, IL-4, IL-6, IL-8, IL-10, and IL-12 genes in intact and hyperplastic endometrium and in the endometrium after hormone therapy were analyzed. Numerous correlations at the proliferation and secretion stages of the menstrual cycle indicate balanced cytokine system. The number of correlations decreases in glandular cystic and more so in atypical hyperplasia, indicating imbalance in the cytokine system. Dufastone and zoladex therapy did not lead to recovery of this balance, but higher correlations between the expression of some factors of cell proliferation attest to the beginning of normalization of pathologically changed endometrium.

[Involvement of tetrahydrobiopterin in local change of endothelium-dependent vasorelaxation in pulmonary hypertension]. / Uchastie tetragidrobiopterina v lokal'nom izmenenii éndoteliizavisimogo rasslableniia sosudov pri legochnoi gipertenzii.

A deficiency of tetrahydrobiopterin (BH4), a NO-synthase co-factor, results in reactive oxygen species synthesis by NO-synthase. It leads to disturbances of endothelium-dependent vasorelaxation. We performed our study on the monocrotaline model of pulmonary hypertension. A decrease in endothelium-dependent relaxation was observed only in intrapulmonary arteries of monocrotaline-treated rats. A perfusion of BH4 (0.1 mol/liter) increased significantly endothelium-dependent dilation of hypertensive pulmonary arteries (p < 0.01). But BH4 did not influence the relaxation of systemic vessels and the dilation responses of pulmonary and systemic arteries of control rats. Measuring of superoxide by lucigenin-mediated chemiluminescence showed five-fold O2- production in intrapulmonary arteries of pulmonary hypertensive rats, that was activated by acetylcholine and inhibited by a nonselective NO-synthase blocker (L-NAME). However, activity of NO-synthase measured as [H3]arginine to [H3]citrulline conversion and assessed in pulmonary vessels and aortic tissue, did not differ in control and monocrotaline-treated groups. These data suggest, that there is a local deficiency of BH4--in pulmonary vessels, without significant changes of systemic circulation.

Expression of cytokine genes in adhesions on uterine tubes.

We studied expression of genes of interleukins, interferon-gamma, tumor necrosis factor-alpha, and transforming growth factor-beta(2) in adhesions on uterine tubes. In tubal adhesions the intensity of production of mRNA for proinflammatory cytokines, antiinflammatory cytokine interleukin-10, and regulator of cell proliferation surpassed that in normal tissues by 2.5-7.4, 2.2, and 50.2 times, respectively. Correlations were found between production of mRNA for tumor necrosis factor-alpha and transforming growth factor-beta(2), interleukin-12, and interferon-gamma and interleukin-12 and transforming growth factor-beta(2). Our results suggest that expression of these genes during adhesion formation is regulated by the feedback mechanism.

[Secretory function of the endothelium as a factor of vascular tone regulation in the norm and in cardiovascular pathology]. / Sekretornaia funktsiia éndoteliia kak faktor reguliatsii sosudistogo tonusa v norme i pri patologii serdechno-sosudistoi sistemy.

Endothelin-1 and nitric oxide are the most potent factors of the endothelium-derived substances. The factors play opposite roles in regulation of cardiovascular system, and their interaction underlies the balance of vasoconstrictor and vasodilator influences on vascular tone under normal conditions. In our experiments, changes in endothelin-1 blood concentration were associated with affected production of endogenous nitric oxide. The altered interrelationships between the endothelium-derived vasoactive substances may precede pathological shifts in the cardiovascular system.

Altered endothelin-dependent regulation of blood pressure and vascular tone in stress-sensitive august rats.

The endothelin (ET) system was studied in August rats, with genetically determined high sensitivity to stress, and in control Wistar rats. Radioimmunoassay revealed a significant difference in plasma endothelin-1 (ET-1) levels of August vs Wistar rats (7.1 fmol/ml vs 50.0 fmol/ml). Immobilization of the animals increased these values up to 11.0 fmol/ml and 65.2 fmol/ml, respectively. Elevation of ET-1 was associated with an increase in blood pressure, which was similar in both strains, whereas the heart rate increase was diminished in the stress-sensitive rats. The mixed endothelin-A/endothelin-B- (ET(A)/ET(B)) receptor antagonist PD142893 suppressed stress-induced elevation of blood pressure in August, but not in Wistar rats. In both strains, heart rate responses to stress were insensitive to the ET receptor blockade. Aortic rings of August rats displayed diminished sensitivity to the vasoconstrictor action of ET-1 vs that of Wistar rats (EC50 = 22.1 nM vs 6.3 nM, respectively). In noradrenaline-precontracted tail arteries, 50 nM ET-1 elicited further constriction without any vasodilator effects. ET-1-induced increase in perfusion pressure was greater in tail arteries of Wistar rats. Thus, endogenous ET-1 can play a strain-dependent role in the stress-induced responses of haemodynamics and the alterations in endothelin-dependent regulation may be responsible for the differences in vascular reactivity of Wistar and August rats.