OBJECTIVETo investigate the differential expression profile and bioinformatic analysis of microRNA (miRNA) in human dental pulp cells (DPC) during endothelial differentiation.

METHODSDPC were cultured in endothelial induction medium (50 µg/L vascular endothelial growth factor, 10 µg/L basic fibroblast growth factor and 2% fetal calf serum) for 7 days. Meanwhile non-induced DPC were used as control.Quantitative real-time PCR (qRT-PCR) was applied to detect vascular endothelial marker genes [CD31, von Willebrand factor (vWF) and vascular endothelial-cadherin (VE-cadherin)] and in vitro tube formation on matrigel was used to analyze the angiogenic ability of differentiated cells. And then miRNA expression profiles of DPC were examined using miRNA microarray and then the differentially expressed miRNA were validated by qRT-PCR. Furthermore, bioinformatic analysis was employed to predict the target genes of miRNA and to analyze the possible biological functions and signaling pathways that were involved in DPC after induction.

RESULTSThe relative mRNA level of CD31, vWF and VE-cadherin in the control group were (3.48 ± 0.22) ×10(-4), (3.13 ± 0.31) ×10(-4) and (39.60 ± 2.36) ×10(-4), and (19.57 ± 2.20) ×10(-4), (48.13 ± 0.54) ×10(-4) and (228.00 ± 8.89) ×10(-4) in the induced group. The expressions of CD31, vWF and VE-cadherin were increased significantly in endothelial induced DPC compared to the control group (P < 0.05). For in vitro tube formation assay, tubular structures were formed on the matrigel by differentiated DPC. A total of 47 miRNA were differentially expressed, in which 15 miRNA were up-regulated and 32 miRNAs down-regulated in differentiated DPC compared with the control. Of these, 4 miRNA were confirmed by qRT-PCR. The target genes of differential miRNA were predicted to associate with several biological functions, such as the regulation of transcription, cell motion, blood vessel morphogenesis, angiogenesis and cytoskeletal protein, and signaling pathways including the mitogen-activated protein kinase (MAPK) and the Wnt signaling pathway.

CONCLUSIONSThe differential miRNA expression identified in this study may be involved in governing DPC endothelial differentiation, thus contributing to the future research on regulatory mechanisms in dental pulp angiogenesis.

Antigens, CD ; Cadherins ; Cell Differentiation ; Collagen ; Computational Biology ; Dental Pulp ; metabolism ; Drug Combinations ; Fibroblast Growth Factor 2 ; Humans ; Laminin ; MicroRNAs ; Platelet Endothelial Cell Adhesion Molecule-1 ; biosynthesis ; Proteoglycans ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; Vascular Endothelial Growth Factor A ; Wnt Signaling Pathway ; von Willebrand Factor

Mast cells are well recognized as key cells in allergic reactions, such as asthma and allergic airway diseases. However, the effects of mast cells and TNF-alpha on T-helper type 2 (Th2) cytokine-dependent asthma are not clearly understood. Therefore, an aim of this study was to investigate the role of mast cells on Th2 cytokine-dependent airway hyperresponsiveness and inflammation. We used genetically mast cell-deficient WBB6F1/J-KitW/KitW-v (W/Wv), congenic normal WBB6F1/J-Kit+/Kit+ (+/+), and mast cell-reconstituted W/Wv mouse models of allergic asthma to investigate the role of mast cells in Th2 cytokine-dependent asthma induced by ovalbumin (OVA). And we investigated whether the intratracheal injection of TNF-alpha directly induce the expression of ICAM-1 and VCAM-1 in W/Wv mice. This study, with OVA-sensitized and OVA-challenged mice, revealed the following typical histopathologic features of allergic diseases: increased inflammatory cells of the airway, airway hyperresponsiveness, and increased levels of TNF-alpha, intercellular adhesion molecule (ICAM)-1, and vascular cellular adhesion molecule (VCAM)-1. However, the histopathologic features and levels of ICAM-1 and VCAM-1 proteins in W/Wv mice after OVA challenges were significantly inhibited. Moreover, mast cell-reconstituted W/Wv mice showed restoration of histopathologic features and recovery of ICAM-1 and VCAM-1 protein levels that were similar to those found in +/+ mice. Intratracheal administration of TNF-alpha resulted in increased ICAM-1 and VCAM-1 protein levels in W/Wv mice. These results suggest that mast cells play a key role in a Th2 cytokine-dependent asthma model through production of adhesion molecules, including ICAM-1 and VCAM-1, by liberation of TNF-alpha.
Animals ; Asthma/*immunology/metabolism/pathology ; Blotting, Western ; Bronchoalveolar Lavage Fluid/immunology ; Cytokines/*immunology ; Intercellular Adhesion Molecule-1/biosynthesis ; Lung/immunology/pathology ; Mast Cells/*immunology/metabolism ; Mice ; Ovalbumin ; Th2 Cells/*immunology ; Tumor Necrosis Factor-alpha/*metabolism ; Vascular Cell Adhesion Molecule-1/biosynthesis

OBJECTIVETo study the effect of Wenmaitong (WMT) and its disassembled formulas on the adhesion of monocytes to endothelial cells induced by hyperlipidemic serum to explore the mechanism of WMT on early arteriosclerosis obliterans (ASO).

METHODSSerums containing whole WMT and its disassembled formulas, including the formula consisted of warming Jing and boosting qi part (Wenjin Yiqi, WY) and that of promoting blood circulation part (Huoxue Tongmai, HT), as well as the serum contained high concentration of lipids were prepared conventionally, respectively. The adhesion of monocytes cell strain THP-1 to human umbilical vascular endothelial cells (HUVEC) was determined by rose bengal stain method, and ELISA was used to detect expressions of intercellular adhesion molecule (ICAM-1), vascular cellular adhesion molecule (VCAM-1) and P-selectin on HUVEC surface.

RESULTSWMT could inhibit THP-1 to HUVEC adhesion induced by hyperlipidemic serum, and down-regulate the expression of ICAM-1, VCAM-1, P-selectin on HUVEC surface, the two disassembled formulas could down-regulate different adhesion molecules.

CONCLUSIONOne mechanism of WMT on ASO may be its inhibition on arteriosclerosis by way of down-regulating the expression of vascular endothelial cells adhesion molecules to decrease the adhesion of monocyte to VEC, therefore to inhibit the monocytes migrating into vascular intima to develop foam cells.

Animals ; Cell Adhesion ; drug effects ; Cell Line ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Humans ; Hyperlipidemias ; blood ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Male ; Monocytes ; cytology ; drug effects ; metabolism ; Rabbits ; Serum ; Vascular Cell Adhesion Molecule-1 ; biosynthesis

OBJECTIVETo investigate the effect of effective parts of Zingiber officinal (EPZ) on the adhesion of ECV-304 cells with monocytes cultivated in vitro and on the expression of monocyte chemotactic protein-1 (MCP-1) and adhesion molecules.

METHODThe model of ECV-304 cell oxidative stress injury was established by hydrogen peroxide (H2O2). Then EPZ-contained blood serum was taken as experimental drug. The adherence of monocytes to endothelial cell were measured by method of rose Bengal. The total RNA of cells was extracted. The intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and MCP-1 mRNA expression in cells were detected by RT-PCR. MCP-1 protein expression were detected by ELISA.

RESULTEPZ could decrease the adhesion of monocytes with ECV-304 cells obviously. Meanwhile it could diminish the expression of ICAM-1, VCAM-1 and MCP-1 in injured ECV-304 cells.

CONCLUSIONEPZ could inhibit H2O2-induced ICAM-1, VCAM-1 and MCP-1 expression in ECV-304 and could inhibit the adherence of monocytes to endothelial cell, which may result in the protect effect in endothelial cells.

Animals ; Cell Adhesion ; drug effects ; Cell Line ; Cells, Cultured ; Chemokine CCL2 ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Ginger ; chemistry ; Humans ; Hydrogen Peroxide ; pharmacology ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Monocytes ; cytology ; drug effects ; metabolism ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

OBJECTIVETo study the protecting effect of polygoni multiflori total glycosides (PMTG) on the atherosclerotic lesion formation and the expression of ICAM-1, VCAM-1 in aolipoprotein (apo) E-deficient transgenic mice.

METHODThirty-two female apoE-deficienct mice were randomized into four groups: PMTG high dose group (150 mg x kg x d), low dose group (25 mg x kg x d), atorvastatin positive control group (5 mg x kg x d), and model group. At the end of the tenth week, all mice were killed. The serum levels of Total cholesterol (TC), Triglyceride (TG), High-density lipoprotein-cholesterol (LDL-C) were measured by enzyme dynamics method. Transmission electron microscopy (TEM) were used to observe the morphologic changes of aortic endothelia cell. The expressions of NF-kappaB were studied by SABC immunohistochemistry.

RESULTAs compared with the model control group. (1) PMTG could reduce the levels of serum TC, TG significantly (P < 0.01), and LDL-C level significantly (P < 0.01). (2) It could increase the levels of serum NO and the anti-oxidation capacities significantly (P < 0.01), but reduce the levels of serum MDA significantly (P < 0.01). (3) PMTG could keep the normal morphology of aortic endothelial cell. (4) PMTG could deregulated the expression of NF-kappaB in aortic wall.

CONCLUSIONPMTG could inhibit the occurrence and development of atherosclerotic lesions by its anti-oxidation abilities, which reduce LDL-C level. The low LDL-C level could deregulated the of expression of NF-kappaB, which could deregulated ICAM-1 and VCAM-1 in AopE-/-mice in aortic wall through.

Animals ; Antioxidants ; pharmacology ; Aorta, Thoracic ; drug effects ; metabolism ; pathology ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; blood ; pathology ; prevention & control ; Cholesterol ; blood ; Cholesterol, LDL ; blood ; Endothelial Cells ; drug effects ; pathology ; ultrastructure ; Female ; Glycosides ; isolation & purification ; pharmacology ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Malondialdehyde ; blood ; Mice ; Mice, Knockout ; Microscopy, Electron, Transmission ; NF-kappa B ; metabolism ; Nitric Oxide ; blood ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Random Allocation ; Triglycerides ; blood ; Vascular Cell Adhesion Molecule-1 ; biosynthesis

OBJECTIVETo explore the effect of hemangiopoietin (HAPO) on the adhesive properties of human umbilical vein endothelial cells (HUVEC).

METHODSThe adhesion of HUVEC and the expressions of CD54, CD102, CD106, CD31, CD62E, and CD62P were measured by adhesion assay, flow cytometry, and semi-quantitative RT-PCR.

RESULTSHAPO enhanced the total adherence of HUVEC in a concentration-dependent manner. Flow cytometry analysis revealed that the treatment of HAPO resulted in a significantly increased expression of CD106 and CD62E on HUVEC in a time-dependent manner. When HUVEC were incubated with HAPO for 6 h, the percentage of CD106 + HUVEC and CD62E HUVEC increased about 2.10 folds and 5.84 folds, respectively, compared with control. The time-course of adhesive molecules mRNA expression indicated that the expression of CD106 and CD62E reached at the maximum 1.86 folds and 6.16 folds, respectively, compared with control.

CONCLUSIONHAPO may facilitate the homing of hematopoietic stem/progenitor cells.

Cell Adhesion Molecules ; metabolism ; Cells, Cultured ; E-Selectin ; biosynthesis ; genetics ; Endothelial Cells ; cytology ; Humans ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Proteoglycans ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

OBJECTIVETo study the protective effect of Naomaitong on inflammatory cascade response after focal cerebral ischemia reperfusion in aged rats.

METHODWe duplicated focal cerebral ischemia model with MCAO, with ischemia 3 h and I/R 1, 3, 6, 12 d points. The effect of Naomaitong on the nervous dysfunction score, the water content of cerebral constitution and the expression of TNF-alpha, VCAM-1, ICAM-1 and its mRNA were observed, and the group with nimodipine was as control.

RESULTThe nervous dysfunction score (I/R1, 3, 6 d), the water content of cerebral constitution (all the time points), the expression of TNF-alpha, VCAM-1 (I 3 h, I/R 1, 3, 6 d), ICAM-1 (I 3 h,I/R 1, 3, 6 d) and its Mrna (all the time points) in model group were higher than those of the sham-operated group; The nervous dysfunction score, the water content of cerebral constitution (I/R 3, 6, 12 d), the expression of TNF-alpha, VCAM-1 (I 3 h, I/R 1, 3 d), ICAM-1 and its mRNA (I 3 h, I/R 1, 3 d) in model group were decreased compared with that of model group. The nervous dysfunction score (I/R 6, 12 d), the expression of TNF-alpha, ICAM-1 (I/R 1d) and its mRNA (I/R 1, 3 d) in Naomaitong group were higher than that of Nimodipine group.

CONCLUSIONNaomaitong could protect brain cell from damage after focal cerebral ischemia reperfusion injury by inhibiting the expression of TNF-alpha, adhesion molecule.

Animals ; Brain ; metabolism ; pathology ; Brain Ischemia ; etiology ; metabolism ; pathology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Infarction, Middle Cerebral Artery ; chemically induced ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Male ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

The aim of this study was to evaluate the effect of all-trans retinoic acid (ATRA) on cell adhesion molecule expression and adhesion capacity of bone marrow stromal cells (BMSC) in patients after conditioning treatment for peripheral blood stem cell transplantation (PBSCT). BMSC of 27 patients before and after conditioning treatment for PBSCT were cultured in vitro. After treated with ATRA at 0.01, 0.1, or 1 micromol/L, expression of intercellular adhesion molecule-1 (ICAM-1) protein and vascular adhesion molecule-1 (VCAM-1) protein were detected by flow cytometry, and soluble ICAM-1 (sICAM-1) protein was determined by using radioimmunoassay. Then BMSC was co-cultured with CD34+ cells, and adhesion rate of BMSC to CD34+ cells was measured. The results showed that after pretreatment with conditioning regimen for PBSCT, the expressions of ICAM-1 and VCAM-1 proteins in BMSC and the expression level of sICAM-1 protein in supernatant of BMSC culture were down-regulated, and the adhesion rate of BMSC to CD34+ cells was decreased, after administration of ATRA, the expression of ICAM-1 protein in BMSC, sICAM-1 protein in culture medium and adhesion rate of BMSC to CD34+ cells all increased significantly, but expression of VCAM-1 protein changed no significantly. It is concluded that the ATRA can partly restore adhesion function of BMSC injured by pretreatment for PBSCT and contribute to hematopoietic reconstitution.
Adolescent ; Adult ; Antigens, CD34 ; Antineoplastic Agents ; pharmacology ; Bone Marrow Cells ; metabolism ; pathology ; Cell Adhesion ; drug effects ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Child ; Coculture Techniques ; Hematologic Neoplasms ; metabolism ; pathology ; therapy ; Humans ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Stromal Cells ; metabolism ; pathology ; Tretinoin ; pharmacology ; Tumor Cells, Cultured ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

This study was aimed to investigate the relationship between endostatin and vascular cell adhesion molecule-1 (VCAM-1) expressions on bone marrow stromal cells (BMSC) in mice after bone marrow transplantation (BMT) and effect of ligustrazine on their expressions. The mice were randomly divided into 3 groups: normal group (without treatment), saline group (control of BMT) and ligustrazine group (BMT + ligustrazine). BMT mouse models were established. The normal group was not treated, the saline group was given normal saline (0.2 ml/mouse, twice a day) through gastric tube, while the ligustrazine group was given ligustrazine (0.2 ml/mouse, twice a day) also through gastric tube. On day 7, 14, 21 and 28 after BMT, mice were killed by euthanasia. The expression levels of endostatin and VCAM-1 in bone marrow stromal cells were detected by immunohistochemistry and RT-PCR analysis respectively. The results showed that the endostatin protein mainly expressed in nuclei of BMSCs, the VCAM-1 protein mainly expressed in plasma of BMSCs. On day 7, 14, 21 after BMT the expression levels of endostatin mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), while their expression levels in ligustrazine group were lower than that in saline group. On day 28 the expression levels in saline group returned to normal, while the expression levels in ligustrazine group not were normalized. On day 7, 14, 21 after BMT the expression levels of VCAM-1 mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), but their expression levels in ligustrazine group were significantly lighter than that in saline group (P < 0.01 or P < 0.05). On day 28 the VCAM-1 expression level in ligustrazine group returned to normal, while its expression level in saline group not were normalized. The difference between these two groups was significant (P < 0.01). Correlation analysis revealed that there was a negative correlation between endostatin and VCAM-1 expression in saline group, there was a positive correlation between endostatin and VCAM-1 expression in ligustrazine group. It is concluded that the endostatin can influence hematopoiesis in bone marrow by affecting VCAM-1 expression on BMSC and hindering connection between stromal cells and hematopoietic cells as well as extracellular stroma and hematopoietic cells, while ligustrazine can enhance the adhesion molecule expression on stromal cell surface of bone marrow in BMT-mice, accelerate the homing and proliferation of HSPC in bone marrow after BMT, meanwhile can promote the repair of bone marrow microenvironment, accelerate hematopoietic reconstitution of bone marrow after BMT through feedback regulation of endostatin expression of BMSC in BMT-mice.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Bone Marrow Transplantation ; Endostatins ; biosynthesis ; genetics ; Female ; Male ; Mice ; Mice, Inbred BALB C ; Pyrazines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Stromal Cells ; metabolism ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

OBJECTIVETo investigate the effects of oxidized low-density lipoprotein receptor 1 (LOX-1) on secretion of adhesive molecules mediated by ox-LDL in human umbilical endothelial cells (HUVECs).

METHODSHUVECs with different concentration of ox-LDL (0, 10, 20, 50, 100 microg/ml) were incubated for 24 h, or HUVECs were pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for another 24 h. Expression of LOX-1 was determined by realtime RT-PCR and Western blot. mRNA and protein of ICAM-1, VCAM-1 and E-selectin were examined by RT-PCR and Western blot respectively.

RESULTSIncubation of HUVECs with ox-LDL (10-100 microg/ml) enhanced the expressions of LOX-1, ICAM-1 and E-selectin in a concentration-dependent manner (P < 0.01). On the contrary, ox-LDL did not affect the expression of VCAM-1 by HUVECs. The expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL were reduced in HUVECs pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for 24 h. This showed that both poly (I) and carrageenan obviously decreased the expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL.

CONCLUSIONox-LDL may upregulate the expression of LOX-1, ICAM-1 and E-selectin, and LOX-1 blocker may partly inhibit this upregulation. The results suggest that the expression of inflammatory molecules induced by ox-LDL in HUVECs is mediated by LOX-1.

Cell Adhesion ; Cell Adhesion Molecules ; Cells, Cultured ; E-Selectin ; metabolism ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipoproteins, LDL ; biosynthesis ; RNA, Messenger ; metabolism ; Receptors, Oxidized LDL ; metabolism ; Scavenger Receptors, Class E ; metabolism ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; metabolism