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1.
National Journal of Andrology ; (12): 428-432, 2010.
Artigo em Chinês | WPRIM | ID: wpr-295046

RESUMO

<p><b>OBJECTIVE</b>To determine the presence of membrane testosterone receptors in cultured vascular smooth muscle cells (VSMC), and investigate their relationship with classical intracellular androgen receptors (iAR).</p><p><b>METHODS</b>VSMCs were cultured from the thoracic aorta of male Sprague-Dawley rats by the explant method. Subconfluent VSMCs were incubated with serum-free medium for 24 h to obtain quiescent non-dividing cells, and then treated with the indicated agents. The aliquots of VSMCs were labeled with testosterone-BSA-FITC (T-BSA-FITC) and analyzed by flow cytometry. Classical iARs in intact- and permeabilized-cells were detected with anti-iAR antibodies and FITC-labeled secondary antibodies by immunofluorescence, followed by flow cytometry analysis.</p><p><b>RESULTS</b>Incubation of VSMCs with T-BSA-FITC obviously increased their relative fluorescence intensity at 10 sec as compared with the untreated controls (P < 0.01), and so did it at 10 min in comparison with the treatment with BSA-FITC alone or together with free testosterone (P < 0.01). Pretreatment with iAR antagonist flutamide exhibited no significant influence on the relative fluorescence intensity of VSMCs (P = 0.318). Traditional iARs were not detectable on the surface of intact VSMCs, although permeabilized cells contained iARs.</p><p><b>CONCLUSION</b>VSMCs contain testosterone receptors in the plasma membrane, and these membrane receptors are not identical to classical iARs.</p>


Assuntos
Animais , Masculino , Ratos , Células Cultivadas , Proteínas de Membrana , Metabolismo , Músculo Liso Vascular , Metabolismo , Ratos Sprague-Dawley , Receptores Androgênicos , Metabolismo , Testosterona , Metabolismo
2.
National Journal of Andrology ; (12): 326-330, 2009.
Artigo em Chinês | WPRIM | ID: wpr-292377

RESUMO

<p><b>OBJECTIVE</b>To explore the acute effects of testosterone at the physiological level on PGF2alpha-induced increase in intracellular Ca2+ in cultured vascular smooth muscle cells (VSMCs).</p><p><b>METHODS</b>VSMCs from the thoracic aorta of male Sprague-Dawley rats were cultured using the explant method. The subconfluent VSMCs were incubated with serum-free medium for 24 hours to obtain quiescent non-dividing cells and then treated with the indicated agents. For the measurement of [Ca2+]i, the VSMCs were loaded with fura-2. Changes of [Ca2+]i were determined ratiometrically with a Nikon TE-2000E system.</p><p><b>RESULTS</b>The resting level of [Ca2+]i was around 100 nmol/L in the VSMCs. Exposing cells to perfusate containing 10 micromol/L PGF2alpha triggered an immediate and transient peak in [Ca2+]i, which gradually decreased afterwards. Interference at the peak with the physiological concentration (40 nmol/L) of testosterone significantly decreased the peak-to-baseline time of [Ca2+]i, compared with ethanol vehicle (104.9 +/- 27.0 s vs 153.5 +/- 40.4 s, P < 0.01). Pretreatment with testosterone at 40 nmol/L for 2 minutes also reduced the peak-to-baseline time of [Ca2+]i significantly in comparison with the ethanol control (120.6 +/- 32.0 s vs 151.4 +/- 27.4 s, P < 0.01), but it had no significant effect on the peak level of PGF2alpha-induced intracellular Ca2+ (390.0 +/- 126.0 nmol/L vs 403.4 +/- 160.7 nmol/L, P > 0.05).</p><p><b>CONCLUSION</b>Testosterone at physiological concentration inhibits PGF2alpha-induced Ca2+ fluxes, probably via receptor-operated calcium channels by a non-genomic mechanism in VSMCs, which may be involved in the vasodilatory effect of testosterone.</p>


Assuntos
Animais , Masculino , Ratos , Cálcio , Metabolismo , Células Cultivadas , Dinoprosta , Farmacologia , Músculo Liso Vascular , Biologia Celular , Miócitos de Músculo Liso , Metabolismo , Ratos Sprague-Dawley , Testosterona , Metabolismo , Fisiologia
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