AIMTo study the co-localization of glucocorticoid receptor and 11beta-hydroxysteroid dehydrogenase 1 (11beta-HSD1) and to investigate whether glucocorticoids regulate the reductase activity and expression of 11beta-HSD1.

METHODSImmunohistochemical staining for 11beta-HSD1 in cultured primary human chorionic trophoblasts was performed. Radiometric conversion assay and Northern blot analysis were respectively used to observe the activity and mRNA expression of 11beta-HSD1.

RESULTS11beta-HSD1 and GR were co-expressed in the same chorionic trophoblast. Both 11beta-HSD1 reductase activity and mRNA levels were increased by dexamethasone (10(-6) mol/L, 10(-7) mol/L) in the cultured chorionic trophoblasts, and the effects were blocked by GR antagonist RU486 (10(-6) mol/L).

CONCLUSIONBy binding to GR, glucocorticoids induce the expression of 11beta-HSD1 by a possible intracrine mechanism.

11-beta-Hydroxysteroid Dehydrogenase Type 1 ; metabolism ; Cells, Cultured ; Chorion ; drug effects ; metabolism ; Female ; Glucocorticoids ; pharmacology ; Humans ; RNA, Messenger ; genetics ; Up-Regulation ; drug effects

OBJECTIVETo observe the angiogenesis promoting effects of clinical common used Chinese herbal medicines (CHM) for activating blood circulation to remove stasis on chick embryo chorio-allantoic membrane (CAM).

METHODSChicken CAM model was established and mice blood serum containing different kinds of medicines, including Radix Peaoniae rubra, Radix Angelicae sinensis, Flos Carthami, Rhizoma Chuanxiong, Radix Salviae miltiorrhizae, Astragalus membranaceus, and their complex prescriptions, Danggui Buxue Decoction, Xuefu Zhuyu Decoction, Xiongshao Capsule, was applied on it respectively to observe the condition of angiogenesis 72 hrs after incubation. Besides, the normal saline group, blank serum group, blank group and basic fibroblast growth factor (bFGF) group were set up for control.

RESULTSAll the CHM applied and bFGF had the CAM angiogenetic promoting effect, among them, Radix Salviae Miltiorrhizae and the three complex prescriptions showed better effects than the three negative control groups in capillary formation and count, with the efficacy similar to that of bFGF. The effect of complex prescriptions was superior to that of single herb except Radix Salviae miltiorrhizae.

CONCLUSIONRadix Salviae miltiorrhizae, Danggui Buxue Decoction, Xuefu Zhuyu Decoction and Xiongshao Capsule have good angiogenesis promoting effect on CAM. This study elucidated, from a certain aspect, the mechanism of action of CHM on ischemic diseases, and unfolded the scientific evidence of applying complex prescription.

Angiogenesis Inducing Agents ; pharmacology ; Animals ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Chorion ; blood supply ; Drugs, Chinese Herbal ; pharmacology ; Mice ; Neovascularization, Physiologic ; drug effects ; Random Allocation ; Salvia miltiorrhiza

Angiostatin is a potent angiogenesis inhibitor that is composed of the first four kringles of plasminogen fragment. Angiostatin with one less kringle molecule (kringle 1 to 3) was recently demonstrated to be an effective angiogenic inhibitor. To determine whether recombinant plasminogen kringle 1-3 (rPK1-3) can inhibit the corneal neovascularization induced by potent angiogenic factors; angiogenin, bFGF, or VEGF, hydron polymer discs each containing 2.0 microg of angiogenin, 500 ng of bFGF, or 500 ng of VEGF respectively were implanted into the corneal stroma of 138 rabbit eyes, and then discs each containing 10 microg, 12.5 microg, 20 microg or 30 microg of rPK1-3 were implanted randomly. Discs containing phosphate buffered saline were also implanted as a control. The angiogenesis score on number and length of newly formed vessels on the each of the rabbit's cornea were recorded daily by two observers (blinded). The treated corneas were also examined histologically. Recombinant PK1-3 treated corneas showed less neovascularization induced by all angiogenic factors (p < 0.05). and the extent of inhibition of neovascularization was proportional to the concentration of rPK1-3 (p < 0.05). Histologic examination showed leukocyte infiltration into the corneal stroma on the PBS treated eyes whereas rPK1-3 treated eyes showed only traces of leukocytes. These results of the effective rPK1-3 inhibition of corneal neovascularization induced by angiogenin, bFGF, or VEGF suggest that this angiostatin related fragment, rPK1-3, may be useful in the treatment of various neovascular diseases. Copyright 2000 Academic Press.
Angiogenesis Inhibitors/pharmacology* ; Angiogenesis Inhibitors/genetics ; Animal ; Chick Embryo ; Chorion/drug effects ; Chorion/blood supply ; Cornea/pathology ; Cornea/drug effects ; Cornea/blood supply* ; Endothelial Growth Factors/pharmacology ; Fibroblast Growth Factor, Basic/pharmacology ; Kringles/genetics ; Lymphokines/pharmacology ; Microscopy/methods ; Neovascularization, Pathologic/drug therapy* ; Plasminogen/pharmacology* ; Plasminogen/genetics* ; Rabbits ; Recombinant Proteins/pharmacology ; Recombinant Proteins/genetics ; Ribonuclease, Pancreatic/pharmacology

Endostatin, a carboxyl-terminal fragment of collagen XVIII is known as an anti-angiogenic agent, that specifically inhibits the proliferation of endothelial cell and the growth of several primary tumor. We report here the purification and characterization of the recombinant murine endostatin (rmEndostatin) which was expressed in a prokaryotic expression system. This rmEndostatin has similar physiochemical properties of yeast-produced recombinant endostatin, and it also specifically inhibits the proliferation and migration of bovine capillary endothelial cells stimulated by basic fibroblast growth factor. The biological activity of rmEndostatin was also shown by its anti-angiogenic ability on the chorioallantoic membrane of chick embryo in vivo. In this article, we demonstrate the refolding and purification of rmEndostatin, expressed using E. coli system, to a biologically active and soluble form. In addition, these results confirm the activity of endostatin as a potent anti-angiogenic agent. Copyright 2000 Academic Press.
Angiogenesis Inhibitors/pharmacology* ; Angiogenesis Inhibitors/isolation & purification ; Angiogenesis Inhibitors/genetics* ; Animal ; Blotting, Western ; Cattle ; Cell Movement/drug effects ; Chick Embryo ; Chorion/pathology ; Chorion/drug effects ; Circular Dichroism ; Collagen/pharmacology* ; Collagen/isolation & purification ; Collagen/genetics* ; Electrophoresis, Polyacrylamide Gel ; Endothelium, Vascular/drug effects ; Endothelium, Vascular/cytology ; Escherichia coli/genetics* ; Fibroblast Growth Factor, Basic/pharmacology ; Mice ; Neovascularization, Physiologic/drug effects ; Peptide Fragments/pharmacology* ; Peptide Fragments/isolation & purification ; Peptide Fragments/genetics* ; Protein Folding ; Recombinant Proteins/pharmacology ; Recombinant Proteins/isolation & purification ; Recombinant Proteins/genetics ; Solubility ; Yeasts/genetics

The aim of this study is to screen out the monoclonal antibody reactive to a kind of endothelial cell line (ECV304) from SZ-1, -2, -21, -22, -51, -65, -262 and explore its function of anti-angiogenesis. The inhibitory effects of monoclonal antibody reactive to ECV304 and human lung carcinom cells (A549) adhesion and migration to extracellular matrix proteins (i.e, fibronectin and collagen IV) were studied by ELISA, the inhibition of angiogenesis in vivo was analyzed by chick chorioallantoic membrane (CAM) assays, the percentage of apoptotic cells in A549 cells was assayed by flow cytometric Annexin-V-FITC/PI dual labeling technique. The results showed that SZ-21 exhibited inhibitory effects on human umbilical vein endothelial cell line (ECV304) and pulmonary cancer cell line (A549) adhesion and migration to extracellular matrix proteins (i.e, fibronectin and collagen IV). In addition, it disrupted ongoing angiogenesis on the chick chorioallantoic membrane (CAM) model. SZ-21 also induced apoptosis of the A549 cells. In conclusion, SZ-21 inhibits angiogenesis in vivo and in vitro by blocking integrin beta(3) and inducing apoptosis. SZ-21 recognized the sequence beta(3) 28 - 35 which is far away from the RGD ligation site on GPIIIa. Integrin may interact the extracellular matrix via recognition sites other than RGD sequence.
Allantois ; blood supply ; Angiogenesis Inhibitors ; pharmacology ; Animals ; Antibodies, Monoclonal ; pharmacology ; Apoptosis ; drug effects ; Cell Adhesion ; drug effects ; Cell Line ; Cell Movement ; drug effects ; Chick Embryo ; Chorion ; blood supply ; Extracellular Matrix Proteins ; pharmacology ; Flow Cytometry ; Humans ; Integrin beta3 ; immunology ; Neovascularization, Physiologic ; drug effects ; Tumor Cells, Cultured