The aim of this study was to elucidate the effects of anti-hypertensive drugs, nifedipine, furosemide, hydrochlorothiazide, captopril, and atenolol on DNA synthesis and proliferation of cultured rat aortic smooth muscle cells induced by fetal calf serum. Aortic smooth muscle cells from Sprague-Dawley rats were isolated, cultured, and seeded in multi-well plates. When confluent, cells were cultured in a conditioned medium without fetal calf serum. After 72 hours, cells were cultured in the medium retaining 10% fetal calf serum with or without anti-hypertensive drugs by increasing the concentration between 10(-8) and 10(-4) M. DNA synthesis was assessed by [3H]-thymidine uptake and proliferation by cell numbers using a hemocytometer. Nifedipine at a concentration of 10(-5) M and 5 x 10(-5) M inhibited serum-induced DNA synthesis significantly by 50.8% and 86.6%, respectively (p < 0.05). The results of cell numbers paralleled those of 3H-thymidine incorporation. Serum-induced DNA synthesis was also reduced by 32.6% at the highest dose of furosemide (10(-4) M), but there was no statistical significance. Hydrochlorothiazide, captopril, and atenolol did not show anti-proliferative effect throughout any of the doses. In conclusion, among the various anti-hypertensive drugs, nifedipine seems to be most beneficial in view of its direct inhibitory effect on DNA synthesis and proliferation of smooth muscle cells, as well as for its anti-hypertensive effect.
Animal ; Antihypertensive Agents/pharmacology* ; Aorta/metabolism* ; Aorta/drug effects* ; Cell Division/drug effects ; Cells, Cultured ; DNA/biosynthesis* ; Male ; Muscle, Smooth, Vascular/metabolism* ; Muscle, Smooth, Vascular/drug effects* ; Muscle, Smooth, Vascular/cytology ; Rats ; Rats, Sprague-Dawley

AIMAccumulated evidence suggest that the development of vascular calcification is similar to osteogenesis. Here we want to elucidate the effect of the common used osteo-regulatory factor 1,25(OH)2D3 on vascular calcification.

METHODS AND RESULTSAdding 10(-9) mol/L to the culture media 1,25(OH)2D3 time dependently increased the calcium deposition on the in vitro calcification of bovine vascular smooth muscle cells (BVSMCs) induced by beta-GP. It also increased cellular alkaline phosphatase activity by 301.1% during the calcified process. Osteocalcin, one of the osteogenic specific metric proteins, was dramatically elevated by 58.3% during the calcified processes, which indicate the transformation of BVSMCs to osteoblastic cell. 1,25(OH)2D3 had no such effect on non-calcified BVSMCs.

CONCLUSIONThese data suggest that 1,25(OH)2D3 exerts a stimulatory effect on vascular calcification through increasing the synthesis of ALP. This effect shares the same character as osteoblast cells. This effect is limited to the calcified prone vascular cell.

Animals ; Calcitriol ; metabolism ; Cattle ; Cells, Cultured ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; pathology ; Osteocalcin ; metabolism ; Vascular Calcification ; metabolism ; pathology ; Vitamin D ; analogs & derivatives ; pharmacology

Animals ; Cell Proliferation ; drug effects ; Female ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tamoxifen ; analogs & derivatives ; pharmacology

OBJECTIVETo study the action mechanism of tetramethylpyrazine (TMP) on the proliferation of vascular smooth muscle cells (VSMCs), thus providing experimental evidence for Chinese medicine to effectively prevent restenosis.

METHODSRats' thoracic aorta VSMCs in vitro cultured (cell line A7r5) were divided into five groups, i.e., the negative control group, the angiotensin II (Ang II, 10(-6) mol/L) group, the low dose TMP (20 micromol/L) plus Ang II group, the middle dose TMP (40 micromol/L) plus Ang II group, the high dose TMP (80 micromol/L) plus Ang II group. The proliferation ratio was detected by MTT. Gene and protein expressions of Wnt4, Dvl-1, beta-catenin, CyclinD1, and collagen I and III were detected with real-time fluorescent quantitative PCR and Western blot respectively.

RESULTSCompared with the negative control group, the proliferation ratio of VSMCs obviously increased in the Ang II group (P < 0.05). Compared with the Ang II group, the proliferation ratio of VSMCs obviously decreased in the middle dose TMP plus Ang II group and the high dose TMP plus Ang II group (P < 0.05). Compared with the negative control group, gene and protein expressions of Wnt4, Dvl-1, beta-catenin, CyclinD1, Col I, and Col III were obviously up-regulated in the Ang II group (P < 0.05). Compared with the Ang II group, mRNA and protein expressions of Wnt4, Dvl-1, beta-catenin, CyclinD1, Col I, and Col III were obviously down-regulated in the middle dose TMP plus Ang II group and the high dose TMP plus Ang II group (P < 0.05). The aforesaid indices were dose-dependent in the low, middle, and high dose TMP plus Ang II groups.

CONCLUSIONTMP inhibited Ang II induced proliferation and collagen secretion of VSMCs through down-regulating Wnt signal pathway.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen ; biosynthesis ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Pyrazines ; pharmacology ; RNA, Messenger ; genetics ; Rats

Oxidized low density lipoprotein (oxLDL) can trigger intracellular production of reactive oxygen species and lipid peroxidation (LPO), and is thought to contribute to initiation and progression of atherosclerosis. In order to understand the correlation between oxLDL and macromolecular damage, we measured levels of LPO-derived miscoding etheno-DNA adducts and LPO-modified proteins in cultured human vascular endothelial and smooth muscle cells after incubation with oxLDL for up to 48 h. A semi-quantative analysis method for 1, N6-ethenodeoxyadenosine (ɛdA) by immunohistochemistry was applied. After oxLDL stimulation, ɛdA-stained nuclei were significantly increased in both endothelial and smooth muscle cells. Similarly, 4-hydroxy-2-nonenal (4-HNE)-modified proteins, as analyzed by immunohistochemistry and Western blotting, were also 3-5 fold increased. It was concluded LPO-derived etheno-DNA adducts and LPO-modified proteins are strongly induced by oxLDL in human vascular endothelial and smooth muscle cells. This macromolecular damage may contribute to the dysfunction of arterial endothelium and the onset of atherosclerosis.
DNA ; metabolism ; Endothelium, Vascular ; cytology ; drug effects ; metabolism ; Humans ; Lipid Peroxidation ; drug effects ; Lipoproteins, LDL ; pharmacology ; Muscle, Smooth ; cytology ; drug effects ; metabolism ; Proteins ; metabolism

Gax gene is a newly found negative transcriptional regulator of cells proliferation. This paper introduces the detection and structural features of Gax, details the inhibitory effect of Gax on the proliferation of vascular smooth muscle cells, vascular endothelial cells and cancer cells, and explains the putative mechanisms therein involved. The potential for providing therapeutic insights into human diseases by modulating Gax activity is prospected.
Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; Gene Expression ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Muscle, Smooth, Vascular ; cytology

Animals ; Cells, Cultured ; Interleukin-1 ; pharmacology ; Matrix Metalloproteinase 1 ; secretion ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Platelet-Derived Growth Factor ; pharmacology ; Rabbits

OBJECTIVETo explore the acute effects of testosterone at the physiological level on PGF2alpha-induced increase in intracellular Ca2+ in cultured vascular smooth muscle cells (VSMCs).

METHODSVSMCs 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.

RESULTSThe 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).

CONCLUSIONTestosterone 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.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Dinoprost ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; metabolism ; Rats ; Rats, Sprague-Dawley ; Testosterone ; metabolism ; physiology

Emodin has such pharmacological effects as ant-inflammatory, anti-tumor, immunoregulation. Meanwhile, emodin could be used for inhibiting the proliferation of vascular smooth muscle cells (VSMC). Many foreign studies demonstrated that emodin had an effect on inhibiting proliferation of VSMCs and cell migration and promoting cell apoptosis, and probed into molecular mechanisms in all aspects. Besides, clinical translational researches and application explorations were also carried out. This article summarizes the research progress of the anti-proliferation effect of emodin on VSMCs.
Apoptosis ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Emodin ; pharmacology ; Humans ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Signal Transduction ; drug effects

AIMTo study the relaxation mechanisms of tetrandrine (Tet) on the corpus cavernosum smooth muscle.

METHODSThe corpus cavernosum smooth muscle cells from New Zealand white rabbits were cultured in vitro. [Ca(2+)](i) was measured by Fluorescence Ion Digital Imaging System, using Fluo-2/AM as a Ca(2+)-sensitive fluorescent indicator.

RESULTSTet (1, 10 and 100 micromol/L) had no effect on the resting [Ca(2+)](i) (P>0.05). In the presence of extracellular Ca(2+) (2.5 mmol/L), Tet (1, 10 and 100 micromol/L) inhibited [Ca(2+)](i) elevation induced by high K(+) and phenylephrine (PE) in a concentration-dependent manner (P>0.05). In calcium free solution containing egtaic acid, Tet (1 and 10 micromol/L) had no inhibitory effects on [Ca(2+)](i) elevation induced by PE (P>0.05). However, Tet (100 micromol/L) inhibited [Ca(2+)](i) elevation induced by PE (P>0.05).

CONCLUSIONTet inhibited the Ca(2+) influx from the extracellular site via voltage-activated Ca(2+) channel and alpha(2)-adrenoceptor-operated Ca(2+) channel. At a high concentration, Tet might inhibit the cytosolic calcium pool release in cultured corpus cavernosum smooth muscle cells. This inhibitory action on [Ca(2+)](i) might be one of the relaxation mechanisms of Tet on the corpus cavernosum smooth muscle.

Alkaloids ; pharmacology ; Animals ; Benzylisoquinolines ; pharmacology ; Calcium ; metabolism ; Cytosol ; drug effects ; metabolism ; Male ; Muscle Relaxation ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Penis ; cytology ; metabolism ; Phenylephrine ; pharmacology ; Potassium Chloride ; pharmacology ; Rabbits