Résumé
<p><b>OBJECTIVE</b>To detect the expression of voltage-gated Na(+) channel (NaCh) isoforms in rat sinoatrial node and explore their functions.</p><p><b>METHODS</b>Expressions of NaCh isoforms Nav1.1, Nav1.2, Nav1.3, Nav1.5, Nav1.6 and Nav1.7 in the rat sinoatrial node were detected by immunohistochemistry. The functional roles of the NaChs were tested by observing the effect of tetrodotoxin, a specific blocker of NaChs, on the intrinsic heart rate of isolated rat working heart.</p><p><b>RESULTS</b>The tetrodotoxin- sensitive neuronal isoforms Nav1.1, Nav1.6 and Nav1.7 as well as the tetrodotoxin-resistant cardiac isoform Nav1.5 were present in the rat sinoatrial node, and the neuronal isoforms were more abundant than Nav1.5 (P<0.05). The selective blockade of tetrodotoxin-sensitive isoforms (presumably Nav1.1, Nav1.6 and Nav1.7) by 100 nmol/L tetrodotoxin scarcely affected the intrinsic heart rate (0.5-/+2.9%, P>0.05) while blockade of tetrodotoxin-resistant isoform (presumably Nav1.5) by 2 micromol/L tetrodotoxin resulted in an obvious decline in the intrinsic heart rate (22.1-/+2.1%, P<0.001).</p><p><b>CONCLUSIONS</b>Nav1.1, Nav1.5, Nav1.6 and Nav1.7 are all present in rat sinoatrial node. Although neuronal isoforms are more abundant, Nav1.5 seems to contribute more to activity of the sinoatrial node.</p>
Sujets)
Animaux , Mâle , Rats , Rythme cardiaque , Physiologie , Immunohistochimie , Ouverture et fermeture des portes des canaux ioniques , Physiologie , Protéines de tissu nerveux , Isoformes de protéines , Noeud sinuatrial , Métabolisme , Physiologie , Canaux sodiques , Tétrodotoxine , PharmacologieRésumé
OBJECTIVE@#To investigate the expression of Kv1.3 and Kir2.1 during human monocyte-derived macrophages differentiation into foam cells and their function in foam cells formation.@*METHODS@#The human macrophage-derived foam cells were obtained by incubating macrophages with ox-LDL (30 mg/L) for 60 h. The expression of Kv1.3 and Kir2.1 channels were examined by immunocytochemistry, RT-PCR and Western blot. Effects of channel blockers (rMargatoxin and BaCl2) on the cellular cholesterol metabolism were studied by measuring the cellular contents of total cholesterol (TC), free cholesterol (FC), and cholesterol ester (CE) in the presence or absence of the channel blockers.@*RESULTS@#After incubating macrophages with 30 mg/L ox-LDL for 60 h, the cellular contents of TC, FC and CE were markedly increased and the ratio of CE/TC was raised from (14.4+/-6.8)% to (57.9+/-3.5)% (P0.05); After being blocked specifically (rMargatoxin: 0.1, 10 nmol/L; BaC(12): 75, 125 micromol/L), the cellular contents of TC and CE were markedly reduced without exception and the ratios of CE/TC were all less than 50% (P<0.05).@*CONCLUSION@#Both Kv1.3 and Kir2.1 channels play a critical role in differentiation of macrophages into foam cells and blockage of corresponding potassium channels would prevent the formation of the foam cells.
Sujets)
Humains , Composés du baryum , Pharmacologie , Différenciation cellulaire , Cellules cultivées , Chlorures , Pharmacologie , Cholestérol ester , Métabolisme , Cellules spumeuses , Biologie cellulaire , Macrophages , Biologie cellulaire , Monocytes , Biologie cellulaire , Canaux potassiques rectifiants entrants , Venins de scorpion , PharmacologieRésumé
A number of mechanical cell stimulators have been used to study the effect of mechanical stimulation on cells in vitro. But the efficiency of these devices is not fully desirable. We recently developed a new device for mechanical cell stimulation, the centrifugal force stretcher, and compared its efficacy with that of the traditional Flexercell Strain Unit. When the mechanical stretcher circumrotates with certain speed, cardiac myocytes attached on the plate are stretched and elongated by centrifugal force. Neonatal rat cardiac myocytes were isolated by enzymatic dissociation from the hearts of 3~5 d old Sprague Dawley rats, and were mechanically stimulated by traditional 20% stretch and 180 r/min centrifugal force for 12 and 24 h. The effects of mechanical stimulation on the hypertrophic response of neonatal rat cardiac myocytes and production of angiotensin II (Ang II) were examined. Compared with the non-stretch group, the radioactivity of (3)H-leucine incorporated into the stretch-stimulated cardiac myocytes in the centrifugal force stretch group was significantly higher [(1295.17+/-51.19) vs (1122.67+/-51.63) in 12 h; (1447.5+/-35.96) vs (1210.67+/-90.92) in 24 h, P<0.05]. Ang II was also dramatically increased by 128% in 12 h (P<0.05) and 139% in 24 h (P<0.01). After the myocytes was stretched for 24 h, the LDH level in the medium in the Flexercell Strain Unit group was significantly higher than that in the centrifugal force group [(14.5+/-8.7) U/L vs (7.8+/-4.3) U/L, P<0.05]. The centrifugal force stretcher is a new and improved mechanical cell stimulator with the same effects on the protein synthesis and Ang II secretion of the cardiac myocytes, and the damage to the cells bronght by this stimulator is relatively slighter in comparison with the Flexercell Strain Unit.