Animals ; Apoptosis/drug effects* ; Autophagy/drug effects* ; Carnitine/pharmacology* ; Cells, Cultured ; Endothelial Cells ; Lipopolysaccharides ; Mice

Endothelial progenitor cells (EPCs) play an important role in diabetic vascular complications. A large number of studies have revealed that some clinical antihyperglycemics can improve the complications of diabetes by regulating the function of EPCs. Metformin can improve EPCs function in diabetic patients by regulating oxidative stress level or downstream signaling pathway of adenosine monophosphate activated protein kinase; Pioglitazone can delay the aging of EPCs by regulating telomerase activity; acarbose, sitagliptin and insulin can promote the proliferation, migration and adhesion of EPCs. In addition to lowering blood glucose, the effects of antihyperglycemics on EPCs may also be one of the mechanisms to improve the complications of diabetes. This article reviews the research progress on the regulation of EPC proliferation and function by antihyperglycemics.
Cell Movement/drug effects* ; Cells, Cultured ; Endothelial Progenitor Cells/drug effects* ; Humans ; Hypoglycemic Agents/pharmacology* ; Signal Transduction/drug effects*

OBJECTIVETo improve the biological properties of decellularized aortic valves by polyethylene glycol (PEG)-mediated covalent incorporation of vascular endothelial growth factor (VEGF).

METHODSPEG crosslinking of decellularized aortic valves were completed via a Michael-type addition reaction, followed by covalent incorporation of VEGF through another Michael-type addition reaction between the unsaturated propylene acyl of PEG and the thiol groups on cysteine residues of VEGF. The effect of VEGF incorporation was evaluated by enzyme-linked immunosorbent assay (ELISA) and immune fluorescence assay. The endothelial progenitor cells (EPCs) were seeded on decellularized aortic valves with or without these modifications, and after 10 days of culture, the valves were examined for DNA content and by hematoxylin-eosin staining and scanning electron microscopy.

RESULTSImmune fluorescence and ELISA showed that the maximal VEGF incorporation on the decellularized aortic valve reached 908.94∓0.27 pg. Compared with the unmodified valves and the valves with PEG crosslinking, decellularized aortic valves with covalent incorporation of VEGF significantly promoted the adhesion and proliferation of EPCs, which formed a confluent cell monolayer on the valve surface.

CONCLUSIONSPEG-mediated covalent incorporation of VEGF in the decellularized aortic valves improves the adhesion and proliferation of the seeded EPCs to facilitate the construction of tissue-engineered heart valves.

Animals ; Aortic Valve ; drug effects ; Cell Adhesion ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Heart Valve Prosthesis ; Polyethylene Glycols ; pharmacology ; Stem Cells ; cytology ; drug effects ; Swine ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo evaluate the possible vascular effects of an environment carcinogen benzo(a)pyrene (BaP).

METHODSThe cytotoxicit of BaP and rat liver S9 (0.25 mg/mL)-activated BaP were examined by MTT assay. Thoracic aortic rings were dissected from Sprague-Dawley rats. Contraction of aortic rings was induced by 60 mmol/L KCl or 10(-6) mol/L phenylephrine (PE) in an ex-vivo perfusion system after BaP (100 μmol/L) incubation for 6 h. [Ca(2+)](i) was measured using Fluo-4/AM. For in-vivo treatment, rats were injected with BaP for 4 weeks (10 mg/kg, weekly, i.p.).

RESULTSBaP (1-500 μm) did not significantly affect cell viability; S9-activated BaP stimulated cell proliferation. BaP did not affect the contractile function of endothelium-intact or -denuded aortic rings. BaP did not affect ATP-induced ([Ca(2+)](i)) increases in human umbilical vein endothelial cells. In BaP-treated rats, heart rate and the number of circulating inflammatory cells were not affected. Body weight decreased while blood pressure increased significantly. The maximum aortic contractile responses to PE and KCl and the maximum aortic relaxation response to acetylcholine were significantly decreased by 25.0%, 34.2%, and 10.4%, respectively.

CONCLUSIONThese results suggest, in accordance with its DNA-damaging properties, that metabolic activation is a prerequisite for BaP-induced cardiovascular toxicity.

Animals ; Aorta ; drug effects ; Benzo(a)pyrene ; pharmacology ; Calcium ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Humans ; Male ; Rats ; Vasoconstriction ; drug effects

To improve the function of endothelial progenitor cells (EPCs) is one of the goals in Chinese traditional therapy to treat various cardio-celebrovascular diseases. In the past decades, scholars in the field of traditional Chinese medicine (TCM) have found fifteen active compounds to regulate the function of EPC. These metabolites are extracted from thirteen, plant-based Chinese medicine, with majority of them as potent reductive or oxidative hydrophilic molecules containing phenyl groups. These active compounds either enhance the mobilization of EPC, or inhibit their apoptosis through different signaling pathways. In this review, the molecular structure, biophysical properties, and the plant sources of these active ingredients and their regulatory effects on the function of EPC are summarized, aiming to reveal the modern basis of Chinese medicine for promoting blood circulation and removing blood stasis at the progenitor cell level.
Animals ; Apoptosis ; drug effects ; Cell Movement ; drug effects ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Progenitor Cells ; cytology ; drug effects ; metabolism ; Humans ; Signal Transduction ; drug effects

AIMTo investigate the influence of adenosine on three-dimensional HUVEC tube formation.

METHODSA new three-dimensional culture type was established in which collagen type I was the substance and two layers of HUVEC grew upward and downward separately. In control group no adenosine was added in the holes; in experiment group 10(-4) mol/L adenosine was added in each hole. Under inverted phase contrast microscope, defined sights were observed and the number of sprout was recorded. Observation lasted for 96 hours, and experiments were repeated 3 times.

RESULTSHUVEC grew upward and downward into collage gel respectively. Tube structure and three-dimensional network was built up gradually. In experiment group(adenosine 10(-4) mol/L) HUVEC grew fast with quick infiltration and sprouting. The tube formation was thick and might form three-dimensional network throughout the collagen gel. At each time point, the difference between experiment and control group was significant (P < 0.05 or P < 0.01).

CONCLUSIONAdenosine may promote HUVEC sprouting and tube-formation.

Adenosine ; pharmacology ; Cells, Cultured ; Cellular Structures ; drug effects ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; Humans

OBJECTIVETo evaluate the effect of lipo-sodium morrhuate on ECV-304 cell line.

METHODSThe effect lipo-sodium morrhuate was evaluated by toxicology trial (MTT), electron microscope, DNA electrophoresis and flow cytometer.

RESULTSThe toxicology results showed, that the number of vital cells in lipo-sodium morrhuate group decreased slowly. The electron microscope exhibited apoptosis in the lipo-sodium morrhuate group. And there were typical DNA ladder in DNA electrophoresis and typical apoptosis peak in flow cytometer. The apoptosis rate was 22.23%.

CONCLUSIONSUnlike the normal preparation of sodium morrhuate, lipo-sodium morrhuate could induce apoptosis of ECV-304 cell line.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; pathology ; Humans ; Liposomes ; Sodium Morrhuate ; pharmacology ; Umbilical Veins ; cytology

BACKGROUNDPrevious studies have shown that resveratrol increases endothelial progenitor cell (EPC) numbers and functional activity. Increased EPC numbers and activity are associated with the inhibition of EPC senescence. In this study, we investigated the effect of resveratrol on the senescence of EPCs, leading to potentiation of cellular function.

METHODSEPCs were isolated from human peripheral blood and identified immunocytochemically. EPCs were incubated with resveratrol (1, 10, and 50 µmol/L) or control for specified times. After in vitro cultivation, acidic β-galactosidase staining revealed the extent of senescence in the cells. To gain further insight into the underlying mechanism of the effect of resveratrol, we measured telomerase activity using a polymerase chain reaction (PCR)-enzyme-linked immunosorbent assay (ELISA) technique. Furthermore, we measured the expression of human telomerase reverse transcriptase (hTERT) and the phosphorylation of Akt by immunoblotting.

RESULTSResveratrol dose-dependently inhibited the onset of EPC senescence in culture. Resveratrol also significantly increased telomerase activity. Interestingly, quantitative real-time PCR analysis demonstrated that resveratrol dose-dependently increased the expression of the catalytic subunit, hTERT, an effect that was significantly inhibited by pharmacological phosphatidylinositol 3-kinase (PI3-K) blockers (wortmannin). The expression of hTERT is regulated by the PI3-K/Akt pathway; therefore, we examined the effect of resveratrol on Akt activity in EPCs. Immunoblotting analysis revealed that resveratrol led to dose-dependent phosphorylation and activation of Akt in EPCs.

CONCLUSIONResveratrol delayed EPCs senescence in vitro, which may be dependent on telomerase activation.

Cells, Cultured ; Cellular Senescence ; drug effects ; Endothelial Cells ; cytology ; drug effects ; enzymology ; Humans ; Stem Cells ; cytology ; drug effects ; enzymology ; Stilbenes ; toxicity ; Telomerase ; metabolism

OBJECTIVETo explore the effect of N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide(W7) on the differentiation from human adipose-derived mesenchymal stem cells (hADSCs) to endothelial cells.

METHODShADSCs were cultured with serum-free differential medium containing 40 ng/ml vascular endothelial growth factor (VEGF) and 10ng/ml basic fibroblast growth factor (bFGF). Cells were divided into control group (differential medium without W7), high-dose group (containing 30 μmol/L W7), medium-dose group (containing 20 μmol/L W7), and low-dose group ( containing 10 μmol/L W7). The hADSCs were cultured for 8 days, and then the changes in the phenotypes of von Willebrand factor (vWF) and vessel-selective cadherin (VE-Cadherin) were detected by flow cytometry (FCM). The intracellular Ca(2+) labeled with Fluo-3 was detected by laser confocal microscopy. After hADSCs planting on Matrigel, their angiogenic potentials were observed under the inverted phase contrast microscope, and the expression of extracellular regulated kinase (ERK) and phosphorylated extracellular regulated kinase (p-ERK) were evaluated by Western blot.

RESULTSAfter the hADSCs were cultured for 8 days, compared with the control group, the expressions of vWF and VE-Cadherin significantly increased along with the decrease of W7 level and the intracellular Ca(2+) also significantly increased (Pü0.01). Lumina-like vascular structure was formed in W7 treatment groups, but not in the blank control group. Compared with the blank control group, the expression of ERK showed no significant in W7 treatment groups (high-, medium-, and low-dose groups)(P>0.05); however, along with the decrease of W7 levels, the expression of p-ERK significantly increased(P<0.05).

CONCLUSIONW7 in proper levels can effectively induce the differentiation from hADSCs to endothelium by increasing the intracellular Ca(2+) level and thus activating the ERK/MAPK pathway.

Adipose Tissue ; cytology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Sulfonamides ; pharmacology

To study the protective effect of combined administration of active ingredients of Danhong on cultured primary mice's brain microvascular endothelial cells (rBMECs) injured by hypoxia. Primary mice's brain micro-vascular endothelial cells were cultured to establish the 4 h hypoxia model. Meanwhile, active ingredients (protocatechuic aldehyde, salvianolic acid B, hydroxysafflor yellow A and tanshinol) of Danhong were administered in rBMECs. The non-toxic dosage was determined by MTT. The leakage of lactate dehydrogenase(LDH), cell superoxide dismutase (SOD) activity and MDA level were detected by the colorimetric method. The expressions of ICAM-1, MMP-9, P53 mRNA were detected by RT-PCR method. Changes in rBMECs cell cycle and early apoptosis were detected by flow cytometry. Danhong's active ingredients and prescriptions 1, 2, 3, 7, 8, 9 could be combined to significantly restrain LDH in hypoxic cells supernatant. Prescriptions 1, 2, 3, 7, 8, 9 could significantly enhance SOD activity in anoxic cells; Prescriptions 1, 2, 3, 8, 9 could significantly decrease the MDA level; Prescriptions 1, 2, 6, 7, 9 could significantly inhibit the early rB-MECs apoptosis induced by hypoxia. After hypoxia, the up-regulated P53 mRNA expression could cause retardation in G, phase and promote cell apoptosis. This proved that the regulatory function of P53 gene lay in monitoring of calibration points in G, phase. Prescriptions 1, 2, 5, 6, 7, 8, 9 could significantly down-regulate the P53 mRNA expression; Prescriptions 1, 4, 7, 8, 9 could significantly down-regulate the ICAM-1 mRNA expression; Prescriptions 1, 3, 6, 9 could significantly down-regulate the MMP-9 mRNA expression. The combined administration of Danhong's active ingredients showed a significant protective effect on primary cultured rBMECs injury induced by hypoxia Its mechanism may be related to the enhancement of cellular antioxidant capacity and the inhibition of inflammatory response and cell apoptosis. This study could provide ideas for researching prescription compatibility, and guide the clinical medication.
Animals ; Apoptosis ; drug effects ; Brain ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Endothelial Cells ; drug effects ; Hypoxia ; drug therapy ; Microvessels ; drug effects ; Rats ; Rats, Sprague-Dawley