Migration of vascular smooth muscle cells (VSMCs) is involved in vascular development and various vascular diseases; however, the molecular mechanisms of VSMC migration remain unclear. In this study, we established an inverted coverslip migration assay to study the migratory properties of cultured VSMCs on extracellular matrix. Pulmonary arterial smooth muscle cells (PASMCs) from rats were cultured and identified by immunocytochemistry. Each coverslip with a confluent monolayer of PASMCs was inverted to a larger coverslip which was coated with phosphate buffered saline (PBS, as a control), poly-D-lysine hydrobromide (PDL), laminin or Matrigel. After 24 h of migration over the larger coverslip, PASMCs were fixed, and reliably quantified. The roles and mechanisms of extracellular matrix in PASMC migration were further studied by wound-healing assay and immunocytochemistry. The results showed that: (1) The purity of the cultured PASMCs was (97 ± 3)%. (2) The number of PASMCs on laminin or Matrigel migrating out from the inverted coverslip was significantly increased compared with that on PBS or PDL, and the migratory distance of PASMCs on laminin or Matrigel was significantly farther than that on PBS or PDL. (3) The motility of PASMCs on laminin or Matrigel was significantly higher than that on PBS at 8 h, 12 h and 24 h after wounding, respectively. (4) F-actin staining showed that F-actin was congregated along the brim of the migrating cells from the inverted coverslip, and vinculin (a cell marker of focal adhesion) staining showed that the distribution of vinculin in PASMCs plated on laminin or Matrigel was significantly lower than that on PBS or PDL. These results suggest that the inverted coverslip migration assay is suitable to study VSMC migration, and laminin and Matrigel substrates may promote VSMC migration through inhibiting the formation of focal adhesion and regulating the cytoskeletal proteins.
Actins ; chemistry ; Animals ; Cell Adhesion ; Cell Movement ; Cells, Cultured ; Collagen ; chemistry ; Drug Combinations ; Extracellular Matrix ; chemistry ; Laminin ; chemistry ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Proteoglycans ; chemistry ; Pulmonary Artery ; cytology ; Rats

BACKGROUND: Cicletanine is a new antihypertensive agent, derived from the furopyridine family. It acts directly on vascular smooth muscle by increasing prostacyclin synthesis and decreasing intracytosolic calcium. In order to investigate the efficacy and safety of cicletanine, a clinical study was performed in the patients with mild to moderate essential hypertension. METHOD: The study subject consisted of 30 patients with diastolic blood pressure of 95mmHg~114mmHg(mean age : 55.1+/-7.9 years, 13 males and 17 females). Cicletanine was administrated orally in a daily dose of 100mg Q.D. for 12 weeks after the administration of a placebo for 2 weeks. During cicletanine medication, antihypertensive efficacy, clinical side effects and laboratory changes were monitored. RESULT: Cicletanine decreased mean blood pressure from the baseline value of 123.6+/-3.4mmHg to 108.6+/-7.5mmHg(p<0.001) after 2 weeks, 105.0+/-7.4mmHg after 4 weeks, 103.9+/-6.6mmHg after 6 weeks, 102.5+/-8.9mmHg after 8 weeks, 101.4+/-6.8mmHg after 10 weeks and 99.6+/-6.6mmHg after 12 weeks of medication. There was a highly significant decrease in blood pressure at each of the assessments after 2,4,6,8,10 and 12 weeks of medication when compared to the baseline value(p<0.001). Mean blood pressure after 4 weeks of medication showed a significant decrease when compared to the value after 2 weeks of medication, and the value after 12 weeks of medication showed a significantly decrease when compared to the value after 8 weeks of medication. Heart rate did not change significantly with cicletanine monotherapy for 12 weeks. There was no significant changes in blood chemistry, glucose, lipid and electrolytes. The side effect was pruritus(1 case, 3.3%). CONCLUSION: Cicletanine monotherapy with 100mg once a day regimen was effective and well tolerated in the patients with mild to moderate essential hypertension.
Blood Pressure ; Calcium ; Chemistry ; Electrolytes ; Epoprostenol ; Glucose ; Heart Rate ; Humans ; Hypertension* ; Male ; Muscle, Smooth, Vascular

Animals ; Drugs, Chinese Herbal ; chemistry ; Humans ; Medicine, Chinese Traditional ; Microscopy, Confocal ; Muscle, Smooth, Vascular ; ultrastructure

Lotus( Nelumbo nucifera) is a traditional medicinal plant,and nowadays it is regarded both as medicine and food. It is widespread across China and rich in natural resources. Almost every part of N. nucifera could be used for medical or edible purpose,including seeds( Lianzi),black ripe fruits( Shilianzi),seed coats( Lianyi),green embryos of mature seed( Lianzixin),flowers( Lianhua),stamens( Lianxu),receptacles( Lianfang),leaves( Heye),leaf or flower stalks( Hegeng),leaf bases( Heyedi),rhizomes( Ou) and rhizome nodes( Oujie). Therefore,this plant is praised as a commercial crop with great economic values. Isoquinoline type alkaloids are the main chemical components of lotus. Smooth muscles usually exist in the digestive tract,respiratory tract and vascular,urinary,reproductive and other human systems. Dysfunction of smooth muscle contraction will induce many diseases including hypertension,asthma and gastrointestinal disorder,etc.,and most of current therapeutic strategies rely on relaxation of smooth muscle by drugs.Previous studies have shown that alkaloids of lotus have strong relaxation activity on smooth muscle. The present paper reviews phytochemistry and smooth muscle relaxation activity of 59 isoquinoline alkaloids from N. nucifera through accessing CNKI,PubMed and multiple databases for biomedical sciences.
Alkaloids/pharmacology* ; China ; Humans ; Isoquinolines/pharmacology* ; Muscle Relaxation ; Muscle, Smooth/drug effects* ; Nelumbo/chemistry* ; Phytochemicals/pharmacology* ; Plant Extracts

OBJECTIVETo investigate whether high glucose-induced vascular calcification is associated with WNT signaling pathway.

METHODSAn in vitro model of human vascular smooth muscle cell (VSMC) calcification was induced by exposure of the cells to high glucose. The expressions of WNT signal molecules and bone-related proteins including Cbfa1, Osx, OCN and BMP2 were analyzed with qRT-PCR, and the cell calcification was assessed by alizarin red staining. The effect of Dkk1, a WNT signaling inhibitor, on high glucose-induced cell calcification was tested with alizarin red staining and calcium content analysis.

RESULTSHigh glucose activated WNT signaling pathway in human VSMCs by up-regulating the expressions of WNT signal molecules including Wnt3a, Wnt7a, Fzd4 and Wisp1 mRNA by 1.86, 1.68, 2.1, and 2.3 folds, respectively, and by promoting the phosphorylation of β-catenin (2.70∓0.22, P<0.05), a key mediator of WNT signaling pathway. Inhibition of WNT signaling pathway by Dkk1 attenuated high glucose-induced VSMC calcification and down-regulated the expression of bone-related proteins Cbfa1, Osx, OCN, and BMP2 by (51∓9)%, (58∓11)%, (56∓10)%, and (62∓10)% (P<0.01).

CONCLUSIONWNT signaling pathway is involved in high glucose-induced VSMC calcification.

Cells, Cultured ; Glucose ; chemistry ; Humans ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Phosphorylation ; Up-Regulation ; Vascular Calcification ; Wnt Signaling Pathway

The aim of this study was to investigate the inhibitory effect of heparin/fibronectin (Hep/Fn) complexes on neointimal hyperplasia following endovascular intervention. Hep/Fn complexes were immobilized onto titanium (Ti) surfaces, with subsequent X-ray photoelectron spectroscopy (XPS), Toluidine Blue O (TBO) and immunohistochemistry methods were used to characterize surface properties. Smooth muscle cell (SMC) cultures were used to evaluate the effect of Hep/Fn complexes on SMC proliferation. Results showed that Hep/Fn complexes successfully immobilized onto Ti surfaces and resulted in an inhibition of SMC proliferation. This study suggests that Hep/Fn surface-immobilized biomaterials develop as a new generation of biomaterials to prevent neointimal hyperplasia, particularly for use in cardiovascular implants.
Biocompatible Materials ; Cell Proliferation ; drug effects ; physiology ; Cells, Cultured ; Fibronectins ; chemistry ; pharmacology ; Heparin ; chemistry ; pharmacology ; Humans ; Immobilized Proteins ; chemistry ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; physiology ; Surface Properties ; Titanium ; chemistry ; Umbilical Arteries

OBJECTIVETo explore the protecting mechanism of Yixintong for heart ischemia-reperfusion injury at cellular and subcellular levels, by observing the effects of Yixintong on three kinds of calcium channels.

METHODThe effects of Yixintong on Ca2+ influx on leak calcium channel, receptor-operationg calcium channel (ROC) and pulse-dependent calcium channel (PDC) were observed respectively, by using rat aortic smooth muscle cell and radioactive 45Ca technique.

RESULTYixintong has no effects on leak calcium channel, but can inhibit the Ca2+ influx in ROC and PDC significantly.

CONCLUSIONYixintong can inhibit the Ca2+ influx in slow channel in a dose-dependent manner.

Animals ; Aorta ; cytology ; Calcium Channels ; drug effects ; Crataegus ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Flavonoids ; isolation & purification ; pharmacology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

Seven sinapine analogs (6a-6g) were synthesized using cinnamon acid or benzoic acid and their derivatives as starting materials, which obtained from substituted benzaldehyde and malonate. The structures of target compounds were characterized by IR, 1H NMR and elemental analysis. The effects of compounds 6a-6g on the smooth muscle of intestine isolated from rabbit were studied, and the experimental results showed that compounds 6a, 6d and 6g had diastolic action, while 6f had contractile action.
Animals ; Choline ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; In Vitro Techniques ; Intestines ; physiology ; Molecular Structure ; Muscle Contraction ; drug effects ; Muscle Tonus ; drug effects ; Muscle, Smooth ; physiology ; Rabbits

The preparation and cell transfection of chitosan-DNA nanoparticles were studied. The TFPI (tissue factor pathway inhibitor) or EGFP (enhanced green fluorescent protein) plasmid DNA was encapsulated with chitosan to form gene nanoparticles. The results with TEM showed that the nanoparticles were of sphere shape. The mean diameter of the nanoparticles was 149 nm and the diameter ranged from 80-250 nm, which were measured by the photo related spectrometry (PCS). The encapsulation efficiency of DNA was 96% +/- 1.38% and the DNA content in the nanoparticles was 37% +/- 3.0%. The encapsulated DNA could be protected from the degradation by DNase I. The transfection efficiency of chitosan nanoparticles were about equivalent to that of the LipofectAMINETM reagent. Our results also showed that chitosan nanoparticles were nontoxic to cultured cells.
Cells, Cultured ; Chitosan ; chemistry ; DNA ; chemistry ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Lipoproteins ; genetics ; Muscle, Smooth, Vascular ; chemistry ; metabolism ; Nanoparticles ; chemistry ; Plasmids ; genetics ; Transfection

OBJECTIVETo investigate the effect of P. notoginseng on vascular smooth muscle cell (VSMC) proliferation induced by hyperlipidemia serum.

METHODMTT method was used to investigate the effect of hyperlipidemia serum and hyperlipidemia plus P. notoginseng on VSMC proliferation.

RESULTHyperlipidemia serum could promote VSMC proliferation significantly as compared with the control group (P < 0.05), while hyperlipidemia plus P. notoginseng could weaken this effect significantly (P < 0.05).

CONCLUSIONP. notoginseng can significantly inhibit the VSMC proliferation induced by hyperlipidemia serum.

Animals ; Aorta, Thoracic ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hyperlipidemias ; blood ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Panax notoginseng ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Serum