Migration and proliferation of smooth muscle cells (SMC) were studied in cultures prepared from the aorta of Wistar male rats (170—200 g b.w., 8 weeks old) raised under conventional (CC) or specific pathogen-free (SPF) conditions. In primary cultures, higher movement of cells from explants was found in CC raised donors, namely in samples cultured in serum incomplete medium. In the following subcultures (passage 3—16), the growth curves were steeper and the doubling time shorter in CC type of cultures. The faster growth of SMC population from conventional donors was found to be due to a shorter cell cycle and a higher proportion of dividing cells. As a consequence, the maximum population densities were also higher in the latter type of cultures. The differences in growth, that were dependent on raising conditions, were evident for 16 passages, i.e. 7 months after explantation of cells into culture. The data suggest that breeding conditions may affect the activation of growth of SMC in blood vessels in situ.
Vascular smooth muscle cells (VSMC) display considerable phenotype plasticity which can be studied in vivo on vascular remodeling which occurs during acute or chronic vascular injury. In differentiated cells, which represent contractile phenotype, there are characteristic rapid transient changes of intracellular Ca2+ concentration ([Ca2+]i), while the resting cytosolic [Ca2+]i concentration is low. It is mainly caused by two components of the Ca2+ signaling pathways: Ca2+ entry via L-type voltagedependent Ca2+ channels and dynamic involvement of intracellular stores. Proliferative VSMC phenotype is characterized by long-lasting [Ca2+]i oscillations accompanied by sustained elevation of basal [Ca2+]i. During the switch from contractile to proliferative phenotype there is a general transition from voltagedependent Ca2+ entry to voltage-independent Ca2+ entry into the cell. These changes are due to the altered gene expression which is dependent on specific transcription factors activated by various stimuli. It is an open question whether abnormal VSMC phenotype reported in rats with genetic hypertension (such as spontaneously hypertensive rats) might be partially caused by a shift from contractile to proliferative VSMC phenotype., E. Misárková, M. Behuliak, M. Bencze, J. Zicha., and Obsahuje bibliografii
Exercise training (ET) is well established to induce vascular adaptations on the metabolically active muscles. These adaptations include increased function of vascular potassium channels and enhanced endothelium-dependent relaxations. However, the available data on the effect of ET on vasculatures that normally constrict during exercise, such as mesenteric arteries (MA), are scarce and not conclusive. Therefore, this study hypothesized that 10 weeks of moderate-intensity ET would result in adaptations towards more vasoconstriction or/and less vasodilatation of MA. Young Fischer 344 rats were randomly assigned to a sedentary group (SED; n=24) or exercise training group (EXE; n=28). The EXE rats underwent a progressive treadmill ET program for 10 weeks. Isometric tensions of small (SED; 252.9±29.5 µm, EXE; 248.6±34.4 µm) and large (SED; 397.7±85.3 µm, EXE; 414.0±86.95 µm) MA were recorded in response to cumulative phenylephrine concentrations (PE; 0-30 µM) in the presence and absence of the BKCa channel blocker, Iberiotoxin (100 nM). In another set of experiments, tensions in response to cumulative concentration-response curves of acetylcholine (ACh) or sodium nitroprusside (SNP) were obtained, and pEC50s were compared. Immunoblotting was performed to measure protein expression levels of the BKCa channel subunits and eNOS. ET did not alter the basal tension of small and large MA but significantly increased their responses to PE, and reduced the effect of BKCa channels in opposing the contractile responses to PE without changes in the protein expression level of BKCa subunits. ET also elicited a sizedependent functional adaptations that involved reduced endothelium-independent and endothelium-dependent relaxations. In large MA the sensitivity to SNP was decreased more than in small MA suggesting impaired nitric oxide (NO)-dependent mechanisms within the vascular smooth muscle cells of ET group. Whereas the shift in pEC50 of ACh-induced relaxation of small MA would suggest more effect on the production of NO within the endothelium, which is not changed in large MA of ET group. However, the eNOS protein expression level was not significantly changed between the ET and SED groups. In conclusion, our results indicate an increase in contraction and reduced relaxation of MA after 10 weeks of ET, an adaptation that may help shunt blood flow to metabolically active tissues during acute exercise.
The morphology and proliferation of vascular smooth muscle cells (VSMC) were studied in cultures prepared from the aorta of newborn male and female Wistar rats. The doubling times (DT) of the male-derived population were 16.4 ±0.7 h and 30.0 ±2.2 h in the exponential and post-exponential growth phases, respectively. In the female donor cells, the corresponding DT values were significantly longer, i.e. 21.9 ± 1.8 h and 38.0 ±2.2 h. In addition, the period of growth was shorter in the female-derived cultures. The percentage of 3H-thymidine labelled cells in male cultures was 61.0±3.1, 92.8± 1.9 and 98.7±0.6 % at 2, 27 and 52 h, respectively. In the female-derived populations, only 24.6 ±4.4, 66.1 ±3.8 and 82.8 ±2.0 % of cells were labelled at the corresponding incubation intervals. As a consequence, the final population density in male cultures was 5.6 times higher. In addition, the male-derived VSMC were mainly spindle-shaped and bulgy in appearance while those from female donors were flat and polygonal which means that the cells were adhering to the growth support to a different extent. The study revealed early determination and long-term persistence of lower adhesiveness as well as higher growth potential of male VSMC, i.e. properties which may be of importance for explaining the higher incidence of vascular wall disorders in males.
The aim of this study was to test the hypothesis that vasorelaxing action of vasonatrin peptide (VNP) is due to activation of the large-conductance Ca2+-activated potassium channel (BKCa) via guanylyl cyclase (GC)-coupled natriuretic peptide receptors (NPRs) in vascular smooth muscle cells (VSMCs). Contraction experiments were performed using human radial artery, whereas BKCa current by patch clamp was recorded in cells from rat mesenteric artery. Contractility of rings cut from human radial artery was detected in vitro. As a result, VNP induced a dose-dependent vasorelaxation of human radial artery, which could be mimicked by 8-Br-cGMP, and suppressed by TEA, a blocker of BKCa, HS-142-1, a blocker of GC-coupled NPRs, or methylene blue (MB), a selective inhibitor of guanylyl cyclase. Sequentially, whole-cell K+ currents were recorded using patch clamp techniques. BKCa current of VSMCs isolated from rat mesentery artery was obtained by subtracting the whole cell currents after applications of 10-7 mol/l iberiotoxin (IBX) from before its applications. In accordance with the results of arterial tension detection, BKCa current was significantly magnified by VNP, which could also be mimicked by 8-Br-cGMP, whereas suppressed by HS-142-1, or MB. Taken together, VNP acts as a potent vasodilator, and NPRA/B-cGMP-BKCa is one possible signaling system involved in VNP induced relaxation., J. Yu ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy