Male rats received estradiol benzoate in a long acting microcrystalline suspension (1 mg/rat i.m., twice a week), methylene blue (MB) 0.5 % in the food and the combination of estradiol and MB. After three weeks, MB partially inhibited the growth response of the anterior pituitary to estradiol and it partially inhibited the increase of cAMP content in anterior pituitary. The increase of anterior pituitary cGMP content was not modified by MB, neither the ratio cAMP/cGMP in the anterior pituitary which, however, decreased after estradiol. This decrease was not modified by MB. On the other hand, the prolactin (PRL) increase in the blood after estradiol was inhibited by MB, although the prolactin content in the anterior pituitary was not. Methylene blue alone did not change blood prolactin concentration, but it unexpectedly elevated blood thyroxine levels and this effect was partially inhibited by simultaneous estradiol treatment.
The effect of 4 weeks’ inhibition of NO synthase by nitro-L-arginine methyl ester (L-NAME) on haemodynamic parameters and cGMP and cAMP content was studied in rat tissues. L-NAME in both 20 mg/kg/day and 40 mg/kg/day doses significantly increased systolic blood pressure by 28 % and 30 % and decreased the heart rate by 14 % and 23 %, respectively, after the first week. These changes persisted during the following three weeks. Left ventricular weight/body weight (LVW/BW) ratio was significantly elevated in both L-NAME-treated groups by 19 % and 29 %, respectively. Radioimmunoassay was used to determine the cGMP and cAMP content Cyclic GMP content in animals treated by L-NAME (20 mg/kg/day and 40 mg/kg/day) decreased significantly by 13 % and 22 % in the left ventricle, by 28 % and 62 % in the aorta, by 20 % and 34 % in the brain, and by 10 % and 15 % in the kidney, respectively. On the other hand, the cAMP content increased in both L-NAME treated groups by 8 % and 9 % in the left ventricle, by 28 % and 46 % in the aorta, and by 23 % and 32 % in the brain, respectively. There were no significant changes in kidney cAMP content as compared to control animals. The results suggest a simultaneous decrease of cGMP and increase of cAMP content in the majority of studied tissues during NO-deficient hypertension.
Atrial natriuretic peptide antifibrotic properties are mainly described in cardiac myocytes or in induced cardiac myofibroblasts (Angiotensin II or TGF-β induced differentiation). In the present work, we investigate the effects of ANP/NPRA/cGMP system in modulating rat cardiac fibroblasts function. Cardiac fibroblasts were isolated from adult Wistar male rats and cultured in the presence of serum in order to induce fibroblasts differentiation. Cultures were then treated with ANP (1 μM), 8-Br-cGMP (100 μM) or IBMX (100 μM), a non-specific phosphodiesterases inhibitor. ANP significantly decreased proliferation rate and collagen secretion. Its effect was mimicked by t he c GMP a nalog, w hile c ombining A NP w ith 8 -Br-cGMP did not lead to additional effects. Moreover intracellular cGMP levels were elevated when cells were incubated with ANP confirming that ANP intracellular pathway is mediated by cGMP. Additionally, immunoblotting and immunofluorescence were used to confirm the presence of guanylyl cyclase specific natriuretic peptide receptors A and B. Finally we scanned specific cGMP dependent PDEs via RT-qPCR, and noticed that inhibiting all PDEs led to an important decrease in proliferation rate. Effect of ANP became more prominent after 10 culture days, confirming the importance of ANP in fibroblasts to myofibroblasts differentiation. Uncovering cellular aspects of ANP/NPRA/cGMP signaling system provided more elements to help understand cardiac fibrotic process., M. Moubarak, C. Magaud, Y. Saliba, A. Chatelier, P. Bois, J.-F. Faivre, N. Farès., and Obsahuje bibliografii
Proximal resistance vessels, such as the mesenteric arteries, contribute substantially to the peripheral resistance. The reactivity of resistance vessels to vasoactive substance like natriuretic peptides plays an important role in the regulation of blood pressure. In current study, we investigated the reactivity of mesenteric arteries to atrial natriuretic peptide (ANP), a well known vasodilating factor, in spontaneously hypertensive rats (SHR), as well as the effects of exercise training on it. As a result, ANP-induced vasorelaxation was attenuated in SHR with significantly increased phosphodiesterase type 5 (PDE5), and decreased cGMP/ANP ratio, compared with WKY rats as control. Intriguingly, the decreased reactivity to ANP in SHR was markedly reversed by exercise training. In addition, ANP resistance of in vitro mesenteric arteries was diminished by sildenafil a potent selective inhibitor of PDE5. In conclusion, ANP resistance occurs in resistance vessels of SHR, suggesting predisposition to hypertension, which can be reversed by exercise., Jun Yu, Bing Zhang, Xing-Lu Su, Ru Tie, Pan Chang, Xue-Ce Zhang, Jian-Bang Wang, Ge Zhao, Miao-Zhang Zhu, Hai-Feng Zhang, Bao-Ying Chen., and Obsahuje bibliografii
Intrafollicular luteinization stimulator was shown to be secreted by granulosa cells in culture with stimulatory effects on differentiation of immature granulosa cells. The purpose of this study was to evaluate the role of calcium ions in luteinization stimulator-enhanced luteinization process of granulosa cells. We examined the direct effect of ionophore A23187, voltage-sensitive Ca2+-channel blockers verapamil, nimodipine, nifedipine, niludipine and calmodulin antagonist trifluoroperazine on progesterone and cGMP levels in 3-day culture of small granulosa cells. It was shown that the dihydropyridine derivatives of calcium blocker drugs (nimodipine, nifedipine, niludipine) and calmodulin antagonist (trifluoroperazine) in the micromolar range, significantly suppressed FSH-induced progesterone synthesis and cGMP accumulation in granulosa cells. On the contrary, phenylalkylamine calcium blocker verapamil and calcium ionophore A23187 had different effects on both processes. Calcium ionophore A23187 markedly enhanced cGMP formation, but simultaneously inhibited the FSH-induced progesterone synthesis. Verapamil at lower concentrations (10 jutA) stimulated and at higher concentrations (50 ptA) inhibited the formation of cGMP. To evaluate the role of extra- and intracellular calcium in luteinization stimulator-enhanced progesterone production by small granulosa cells, the effects of indicative agents on stimulatory activity of follicular fluid from large follicles, granulosa cells-conditioned media and granulosa cell extracts were tested. While verapamil is shown to be a less potent modulator, administration of other calcium antagonists as well as ionophore A23187 caused a significant decrease in stimulatory action of follicular fluid from large follicles, granulosa cells-conditioned media and extracts. These findings indicate that the stimulatory action of luteinization stimulator depends on the transport of calcium ions through voltage- sensitive calcium channels and is modulated by alteration of intracellular calcium levels.
Male and female rats were given oestradiol benzoate (1 mg s.c. twice a week for 3 weeks) and/or sodium nitroprusside (SN), a donor of nitric oxide (NO), which was administered in their food in amounts of 0.2 or 0.6 mg/rat/day. Neither oestradiol-induced hypertrophy of the hypophysis, nor the serum prolactin (PRL) level, was affected by the simultaneous administration of SN. The PRL content of the hypophysis rose after oestradiol in the males, but the increase was again uninfluenced by the simultaneous administration of SN and the cAMP content of the hypophysis - raised after oestradiol - was likewise unaffected. The amount of cGMP in the hypophysis after oestradiol rose only in males. Both the serum and the hypophyseal prolactin level were found to be correlated to the cAMP and the cGMP content of the hypophysis. It was found that the simultaneous administration of SN together with oestradiol slightly reduced the increase in the cGMP content of the hypophysis elicited with oestradiol treatment only.