angiogenin and Kidney-Failure--Chronic

Degranulation of polymorphonuclear leukocytes (PMNL) occurs during extracorporeal circulation. A degranulation-inhibiting protein identical to angiogenin was recently isolated from high-flux dialyzer ultrafiltrate. This protein inhibits the release of lactoferrin and metalloproteinases from PMNL in vitro. In the present study, we investigated end-stage renal disease patients undergoing regular hemodialysis treatment with either high-flux dialyzers (n = 51) or low-flux dialyzers (n = 44), and chronically uremic patients undergoing hemodiafiltration (n = 30). Hemodialysis therapy with low-flux polysulfone or cellulose triacetate membranes caused no or only minimal reduction (

Topics: Adult; Aged; Aged, 80 and over; Azo Compounds; Cell Degranulation; Cellulose; Extracorporeal Circulation; Female; Humans; Kidney Failure, Chronic; Lactoferrin; Leukocyte Elastase; Male; Membranes, Artificial; Middle Aged; Neutrophils; Polymers; Renal Dialysis; Ribonuclease, Pancreatic; Sulfones

Experimental data confirmed that erythropoietin (EPO) administration alters the course of various pathological situations such as heart failure and tumor growth by inducing vascular endothelial growth factor-A (VEGF-A) expression. The effect of EPO dose on plasma VEGF-A level in hemodialysis (HD) patients was evaluated. The effect of EPO dose on plasma angiogenin level in HD patients was also evaluated, since angiogenin is necessary for angiogenesis induced by VEGF-A.. Thirty two HD patients (10 diabetics) enrolled into the study. Patients were iron replete and did not suffer from infections, autoimmune diseases or malignancies. Plasma VEGF-A and angiogenin, as well as serum interleukin-6 and tumor necrosis factor-α were measured by means of ELISA.. Weekly EPO dose per kg of dry body weight was positively related to both VEGF-A and angiogenin, whereas no relation was detected among VEGF-A or angiogenin and hemoglobin, inflammation or presence of diabetes mellitus. These relations among EPO dose and VEGF-A or angiogenin remained after adjustment for hemoglobin concentration or inflammation or presence of diabetes mellitus.. EPO dose may affect plasma VEGF-A and angiogenin concentrations in HD patients.

Topics: Aged; Diabetes Complications; Dose-Response Relationship, Drug; Erythropoietin; Female; Gene Expression Regulation; Humans; Inflammation; Interleukin-6; Kidney Failure, Chronic; Male; Middle Aged; Renal Dialysis; Ribonuclease, Pancreatic; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

A degranulation inhibiting protein was purified to apparent homogeneity from plasma ultrafiltrates of patients with uremia using gel-permeation chromatography, ion-exchange chromatography, affinity chromatography on blue Sepharose, and ion-exchange chromatography on a Mono S HR 5/5 fast protein liquid chromatography column. The identity of the isolated degranulation inhibiting protein with angiogenin was demonstrated by amino acid sequence determination, immunoblotting, and identical inhibitory activity effects on leukocyte degranulation. At concentrations in the nanomolar range, the protein inhibited spontaneous degranulation of polymorphonuclear leukocytes (PMNL) to 40%. The protein discharge of cells, which were preincubated with nanomolar concentrations of angiogenin and then stimulated with the chemotactic peptide formyl-norleucyl-leucyl-phenylalanyl-norleucyl-tyrosyl-leucine (FNLPNTL), was inhibited by 70%. Cellular functions such as chemotaxis, phagocytosis, and the oxidative respiratory burst were not obviously affected by angiogenin. A polyclonal antibody to human recombinant angiogenin abolished the inhibitory effect of the isolated protein upon PMNL. The same but reduced effect was induced by the disulfide C39-C92 containing tryptic angiogenin fragment L-H-G-G-S-P-W-P-P-C92-Q-Y-R-G-L-T-S-P-C39-K, indicating a new, so far unknown biologically active site of angiogenin which is different from the sites responsible for angiogenic or ribonucleolytic activity. Two synthetic peptides containing residues 83-95, one with S92 instead of C92, revealed the same inhibitory effect on the protein degranulation of PMNL as the entire tryptic fragment.

Topics: Amino Acid Sequence; Cyanogen Bromide; Cytoplasmic Granules; Electrophoresis, Polyacrylamide Gel; Humans; Kidney Failure, Chronic; Kinetics; Molecular Sequence Data; Neutrophils; Peptide Fragments; Proteins; Reference Values; Ribonuclease, Pancreatic; Trypsin