The expression of endothelial nitric oxide synthase traffic inducer (NOSTRIN) was examined in the umbilical vessels of the patients with pre-eclampsia (PE) to explore its possible role in the pathogenesis of PE. The NOSTRIN mRNA in umbilical tissues was determined by RT-PCR. The eNOS activity in umbilical vessels was spectrophotometrically detected. NO2-/NO3-, the stable metabolic end products of NO, was measured by using nitrate reductase. RT-PCR showed that the expression level of NOSTRIN was significantly higher in women with PE than in the normal group (P<0.01). The activity of eNOS was significantly decreased in PE group [(12.83+/-3.61) U/mg] than in normal group [(21.72+/-3.83) U/mg] (P<0.01). The level of NO2-/NO3- in PE patients (27.53+/-7.48) micromol/mg was significantly lower than that of normal group (54.27+/-9.53) micromol/mg (P<0.01). The significant negative correlation existed between the expression of NOSTRIN and the activity of eNOS in umbilical vessels of women with PE (r=-0.58, P<0.01). It was concluded that the level of NOSTRIN expression was increased in umbilical vessel of women with PE, indicating that it may be involved in the pathogenesis of PE.
Intracellular Signaling Peptides and Proteins/genetics ; Intracellular Signaling Peptides and Proteins/*metabolism ; Pre-Eclampsia/*enzymology ; Pre-Eclampsia/etiology ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Umbilical Arteries/cytology ; Umbilical Arteries/*enzymology ; Umbilical Veins/cytology ; Umbilical Veins/*enzymology

The proliferation difference between Human Umbilical Vein Endothelial Cells (HUVEC) and Human Umbilical Artery Smooth Muscle Cells (HUASMC) in response to the concentration of silver nanoparticles was investigated via MTT, BCA and FCM tests. The obtained experimental data were statistically analyzed and discussed in order to know the causation of the proliferation difference. The results show there is significant difference in proliferation between HUVEC and HUASMC corresponding to the concentration of silver nanoparticles, and such difference can be attributed to the varied adhesion shape and apoptosis of the cells being influenced by nano-Ag content and Fetal bovine serum (FBS) content in culture medium.
Cell Proliferation ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Metal Nanoparticles ; chemistry ; Muscle, Smooth, Vascular ; cytology ; Silver ; pharmacology ; Umbilical Arteries ; cytology

We assessed the effects of pulsatile flow shear stress on the gene expression profiles of human umbilical artery smooth muscle cells (HUASMCs) in vitro using the Express Chip DNA microarray method and investigated the difference between pulsatile and steady shear stress on differentially expressed genes of HUASMCs. In a modified pulsatile flow chamber system, HUASMCs were exposed to pulsatile and steady fluid shear stress (5.52 dyne/cm2) for 6 h respectively, and normal static cultured HUASMCs were selected as a control. The total cellular RNA was extracted by TRIzol Reagent (Life Technologies, Inc) according to the manufacturer's manual. Conversion of mRNA to single strand cDNA and double strand cDNA template was synthesized by Reverse Transcription from the total RNA. cRNA probe was transcribed with biotin labeling. After hybridization of probe with microarray, the binding of streptavidin to biotin was performed and amplified with the first antibody and further amplified with Cy3-conjugated second antibody. Then detection of Cy3 dye was carried out with ScanArray 5000. The results showed that a total of 1,330 genes revealed differential expression in HUASMCs exposed on pulsatile shear stress (5.52 dyne/cm2, 6 h); however, 2,676 genes revealed differential expression in HUASMCs exposed on steady shear stress. Comparsion of HUASMCs exposed to pulsatile with the HUASMCs exposed to steady shear stress showed there were 2,297 genes revealing differential expression. The transcriptional profile of fluidally induced genes in HUASMCs suggested a different response to pulsatile and steady shear stress.
Cells, Cultured ; Gene Expression Profiling ; Humans ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Oligonucleotide Array Sequence Analysis ; Pulsatile Flow ; RNA, Messenger ; biosynthesis ; genetics ; Stress, Mechanical ; Umbilical Arteries ; cytology

OBJECTIVE: To observe the expression of the co-expression plasmid of tissue-plasminogen activator (tPA) and vascular endothelia growth factor165 (VEGF165) in vascular smooth muscle cells (VSMC) and to study the effect of expressing products on the proliferation of VEC and VSMC and fibrinolysis activity. METHODS: The co-expression plasmid of tPA and VEGF165 (pBudCE4.1/tPA-VEGF165) was transfected into VSMC with the lipofection. The expression of tPA and VEGF165 at mRNA level was detected by RT-PCR and the protein level expression was detected by enzyme linked immunosorbent assay (ELISA). The fibrinolysis activity of culture medium of VSMC transfected tPA and VEGF165 genes was detected by fibrin plate technique. The VEC and VSMC were cultured with culture medium of VSMC transfected tPA and VEGF165 genes. And the proliferation of VEC and VSMC was evaluated with monoterazolium (MTT) and flow cytometry (FCM). RESULTS: The expression of tPA and VEGF165 at mRNA and protein levels in the transfected VSMC was demonstrated by RT-PCR and ELISA, respectively. The VSMC culture medium of transfected genes possessed evidently fibrinolysis activity. The expression products of tPA and VEGF165 in the VSMC had an evident effect on the VEC proliferation. But it had not an effect on the VSMC proliferation. CONCLUSION The eukaryotic co-expression plasmid of tPA and VEGF165 can be expressed in transfected VSMCs. The expression products have an obvious biological activity. The present study lay the foundation for future making use of tPA and VEGF165 to prevent and treat vascular stenosis in transplanted heart.
Cell Proliferation ; Cells, Cultured ; Endothelium, Vascular ; cytology ; Fibrinolysis ; Humans ; Muscle, Smooth, Vascular ; cytology ; Plasmids ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Tissue Plasminogen Activator ; biosynthesis ; genetics ; Transfection ; Umbilical Arteries ; cytology ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

OBJECTIVETo study the effect of angiotensin II (Ang II) on pregnancy-associated plasma protein A (PAPP-A) and insulin-like growth factor 1 (IGF-1) mRNA expressions in human umbilical artery smooth muscle cells (HUVSMCs).

METHODSIn the presence or absence of Ox-LDL, HUVSMCs were cultured with Ang II of 10(-5) mol/L for 0, 6, 12, 24, 36, and 48 h, or with Ang II at the concentrations of 10(-7), 10(-6), 10(-5), and 10(-4) mol/L for 24 h, after which the cells were then collected to detect PAPP-A and IGF-1 mRNA expressions in the cells using RT-PCR.

RESULTSAt the concentration of 10(-5) mol/L, Ang II showed a time-dependent effect in inducing PAPP-A and IGF-1 mRNA expressions, which began to increase at 12 h of culture and reaching the highest level at 24 h. Ang II also dose-dependently induced PAPP-A and IGF-1 mRNA expressions, and 10(-5) mol/L Ang II induced the highest expression levels of the two genes. Ox-LDL exposure significantly further increased the expression levels of PAPP-A and IGF-1 mRNA in the cells regardless of the Ang II concentration or duration for cell treatment (P<0.05).

CONCLUSIONAng II can time- and dose-dependently induces PAPP-A and IGF-1 mRNA expression in HUVSMCs and is responsible for inducing platelet activity and inflammatory reaction in acute coronary syndromes, and the effects of Ang II can be enhanced by Ox-LDL.

Angiotensin II ; pharmacology ; Cells, Cultured ; Drug Synergism ; Humans ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Lipoproteins, LDL ; pharmacology ; Myocytes, Smooth Muscle ; cytology ; metabolism ; Pregnancy-Associated Plasma Protein-A ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Umbilical Arteries ; cytology ; metabolism

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 evaluate the effects of rapamycin (RPM)-loaded poly (lactic-co- glycolic) acid (PLGA) nanoparticles (NPs) on the proliferation, distribution of cell cycle, and expression of p27 protein in human umbilical arterial vascular smooth muscle cell (HUASMC) in vitro.

METHODSThe primarily culture model of HUASMC was successfully established by explant-attached method in vitro. The cells were administrated with different doses of RPM, and RPM-PLGA NPs were observed as treat groups compared with PLGA NPs and M231-SMGs medium cultured group. The effect of RPM-PLGA NPs on proliferation of HUASMC was assessed using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) colorimetry method. The influences of RPM-PLGA NPs on the cell cycle and cellular growth kinetics of HUASMCs were tested by flow cytometry. The effect of RPM-PLGA NPs on the expression of p27 protein of HUASMCs was assessed through an immunohistochemical method.

RESULTSCompared with the control group, the proliferation of HUASMCs was inhibited by 50 microg/L and higher concentration of RPM-PLGA NPs in a dose-dependent manner (P < 0.05). The numbers of cells entering cell cycle of S/G2/M phases were significantly lower in RPM-PLGA NPs and RPM treated groups. Histologically, the expression of p27 were up-regulated in 500 microg/L RPM-PLGA NPs and 100 microg/L RPM treated group (all P < 0.01 ) when compared with the control group.

CONCLUSIONSRPM-PLGA NPs has a similar effects as RPM in inhibiting the growth of in vitro cultured HUASMC. It can remarkably suppress the expression of in vitro cultured HUASMC p27 protein, arrest its cell cycle at G1/S phase, and inhibit its proliferation.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Drug Carriers ; Humans ; Lactic Acid ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Nanoparticles ; Polyglycolic Acid ; Sirolimus ; administration & dosage ; pharmacology ; Umbilical Arteries ; cytology

BACKGROUNDIt has been identified that in patients with pre-eclampsia, several factors were released into the serum, which can regulate the proliferation of vascular smooth muscle cells and stimulate the synthesis of extracellular matrix (ECM). However, the signal transduction pathway of the growth factors and cytokines synthesized by endothelial cells leading to the proliferation and ECM synthesis of human umbilical arterial smooth muscle cell (HUASMC) is unknown. The aim of this study was to research the effects of protein kinase C (PKC) inhibitor, polymyxin B (PMB), on the proliferation, nuclear factor-kappa B (NF-kappaB) activation, and expression of procollagen I and III mRNA in HUASMC cultured with pre-eclamptic umbilical sera.

METHODSNormal HUASMCs were treated with pre-eclamptic umbilical serum (pre-eclamptic group), pre- eclamptic umbilical serum plus PMB (PMB group), or normal umbilical serum (normal group). The expression of I-kappaB, NF-kappaB was detected by Western blotting after the HUASMC was incubated for 2 hours. The proliferation of HUASMC was evaluated by MTT, the cell cycle was analyzed by flow cytomerty, and the expression of procollagen I, III mRNA was measured by RT-PCR assay after HUASMC was incubated for 48 hours. ANOVA was used for statistical analysis.

RESULTSUmbilical sera in pre-eclampsia could stimulate the proliferation, the DNA synthesis, the transition of G(0) + G(1) phase or G(2)/M phase to S and phase, the activation of NF-kappaB, and the expression of procollagen I mRNA of HUASMC as compared with normal umbilical sera (P < 0.01). PMB could inhibit the proliferation, the DNA synthesis, the transition of G(0) + G(1) or G(2)/M phase phase to S phase, the activation of NF-kappaB, and the expressions of procollagen I mRNA of HUASMC stimulated by umbilical sera in pre-eclampsia (P < 0.01).

CONCLUSIONPKC-NF-kappaB signal transduction pathway may play a key role in SMC phenotype modulation, which is more important in the pathogenesis of placental blood vessel in pre-eclampsia.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; genetics ; Collagen Type III ; genetics ; Female ; Humans ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; NF-kappa B ; physiology ; Polymyxin B ; pharmacology ; Pre-Eclampsia ; metabolism ; pathology ; Pregnancy ; Protein Kinase C ; antagonists & inhibitors ; physiology ; RNA, Messenger ; analysis ; Signal Transduction ; Umbilical Arteries ; metabolism