NAD(P)H oxidase activity in cultured human podocytes: Effects of adenosine triphosphate

被引:120
作者
Greiber, S
Munzel, T
Kastner, S
Muller, B
Schollmeyer, P
Pavenstadt, H
机构
[1] Univ Freiburg, Med Klin 4, Nephrol Abt, D-79106 Freiburg, Germany
[2] Univ Krankenhaus Eppendorf, Abt Kardiol, Hamburg, Germany
关键词
xanthine oxidase; kidney; oxygen radicals; lucigenin; polymerase chain reaction; adenosine triphosphate;
D O I
10.1046/j.1523-1755.1998.00796.x
中图分类号
R5 [内科学]; R69 [泌尿科学(泌尿生殖系疾病)];
学科分类号
1002 ; 100201 ;
摘要
Reactive oxygen species contribute to glomerular damage and proteinuria. In this study, we show that cultured human podocytes produce superoxide in response to extracellular adenosine triphosphate (ATP), and we identified the oxidases involved in this process. Adenosine triphosphate (10(-4) M far 4 hr) raised superoxide production from 1.28 +/- 0.15 to 2.67 +/- 0.34 nmol/mg protein/min. Studies with podocyte homogenates revealed activation of both nicotinamide adenine dinucleotide (NADH; from 2.65 +/- 0.23 to 7.43 +/- 0.57) and nicotinamide adenine dinucleotide phosphate (NADPH) dependent oxidases [from 1.74 +/- 0.13 to 4.05 +/- 0.12 (nmol O-2(.)/mg protein/min)] by ATP. Activity of xanthine-oxidases was low and unchanged by ATP. Activation of the plasma-membrane bound NAD(P)H oxidases by ATP was time and dose dependent. Reverse transcribed-polymerase chain reaction (RT-PCR) studies with primers derived from monocyte sequences amplified mRNA for the NADPH oxidase subunits p22phox, p47phox, gp91phox and p67phox, and the latter was transiently increased by ATP. Experiments with actinomycin D and cycloheximide suggested that ATP modulates enzyme activity at the transcriptional and translational levels. In conclusion, NAD(P)H dependent, membrane associated oxidases represent the major superoxide source in human podocytes. Activation of NAD(P)H oxidase by ATP might be secondary to increased mRNA expression of the NADPH oxidase subunit gp67phox.
引用
收藏
页码:654 / 663
页数:10
相关论文
共 51 条
  • [1] ACTIVATION OF THE NADPH OXIDASE INVOLVES THE SMALL GTP-BINDING PROTEIN P21RAC1
    ABO, A
    PICK, E
    HALL, A
    TOTTY, N
    TEAHAN, CG
    SEGAL, AW
    [J]. NATURE, 1991, 353 (6345) : 668 - 670
  • [2] Mouse vitamin D-24-hydroxylase: Molecular cloning, tissue distribution, and transcriptional regulation by 1 alpha,25-dihydroxyvitamin D-3
    Akeno, N
    Saikatsu, S
    Kawane, T
    Horiuchi, N
    [J]. ENDOCRINOLOGY, 1997, 138 (06) : 2233 - 2240
  • [3] ASSEMBLY AND REGULATION OF NADPH OXIDASE AND NITRIC-OXIDE SYNTHASE
    BASTIAN, NR
    HIBBS, JB
    [J]. CURRENT OPINION IN IMMUNOLOGY, 1994, 6 (01) : 131 - 139
  • [4] Brandes Ralf P., 1996, Journal of the American Society of Nephrology, V7, P1653
  • [5] A GROWTH FACTOR-STIMULATED AND HORMONE-STIMULATED NADH OXIDASE FROM RAT-LIVER PLASMA-MEMBRANE
    BRIGHTMAN, AO
    WANG, J
    MIU, RKM
    SUN, IL
    BARR, R
    CRANE, FL
    MORRE, DJ
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1992, 1105 (01) : 109 - 117
  • [6] CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2
  • [7] A ROLE FOR OXYGEN FREE-RADICALS IN AMINONUCLEOSIDE NEPHROSIS
    DIAMOND, JR
    BONVENTRE, JV
    KARNOVSKY, MJ
    [J]. KIDNEY INTERNATIONAL, 1986, 29 (02) : 478 - 483
  • [8] HORMONAL MODULATION OF GLOMERULAR FUNCTION
    DWORKIN, LD
    ICHIKAWA, I
    BRENNER, BM
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1983, 244 (02): : F95 - F104
  • [9] Identification and functional characterization of a calcium channel gamma subunit
    Eberst, R
    Dai, SP
    Klugbauer, N
    Hofmann, F
    [J]. PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1997, 433 (05): : 633 - 637
  • [10] PUROMYCIN AMINONUCLEOSIDE METABOLISM BY GLOMERULI AND GLOMERULAR EPITHELIAL-CELLS INVITRO
    GHIGGERI, GM
    CERCIGNANI, G
    GINEVRI, F
    BERTELLI, R
    ZETTA, L
    GRECO, F
    CANDIANO, G
    TRIVELLI, A
    GUSMANO, R
    [J]. KIDNEY INTERNATIONAL, 1991, 40 (01) : 35 - 42