PMID- 11087070
OWN - NLM
STAT- MEDLINE
DCOM- 20010222
LR  - 20171116
IS  - 0255-6596 (Print)
IS  - 0255-6596 (Linking)
VI  - 24
IP  - 4
DP  - 2000 Oct
TI  - An increase in free radical production by means of an anion channel blocker DIDS 
      in mouse peritoneal neutrophils.
PG  - 178-86
AB  - DIDS (4, 4'-diisothiocyanostilbene-2, 2'-disulfonic acid) has been recognized as 
      an anion channel blocker. In this study, we demonstrated that DIDS significantly 
      enhanced the production of free radicals in mouse peritoneal neutrophils. By 
      means of a luminol-chemiluminescence (LCL) monitoring system, DIDS markedly 
      increased LCL which could be suppressed by SOD, sodium azide (NaN3), EGTA and 
      BAPTA-AM and only slightly inhibited by staurosporine (STP). Depletion of the 
      endoplasmic reticulum (ER)-Ca2+ store by means of thapsigargin (TG) had no 
      effects on DIDS-enhanced LCL, but DIDS significantly increased the amount of 
      intracellular free calcium as monitored by means of fura-2 staining. These 
      results indicate that DIDS may enhance free radical production mediated by Ca2+ 
      release from the mitochondria. Both phorbol-12-myristate-13-acetate (PMA) and 
      DIDS can induce increased translocation of p47-phox of the neutrophil to the 
      membrane fraction, which is inhibited by STP pretreatment. Since free radical 
      generation could reduce the cytoplasmic pH (pHi), we further examined whether 
      DIDS was capable of inducing intracellular acidification. The result indicated 
      that DIDS certainly lowered the pHi which was also suppressed by pretreatment 
      with either NaN3 or NaCN, but not by diphenyleneiodonium (DPI). These findings 
      lead us to propose a working hypothesis that DIDS mainly induces superoxide 
      production accompanied by decreasing pHi mediated through a Ca2+ -dependent 
      effect on the mitochondria rather than on NADPH oxidase. Using the lipophilic 
      fluorescent dye DiOC6(3), we showed that DIDS decreased the transitional 
      mitochondrial membrane potential. NaN3, but not STP or pyrrolidine 
      dithiocarbamate (PDTC), antagonized DIDS in the course of decreasing the 
      mitochondrial membrane potential. Taken together, all of these findings imply a 
      possible role of anion channels of the mitochondria in modulating free radical 
      production and intracellular acidification of neutrophils through alteration of 
      the mitochondrial transition membrane potential and Ca2+ -release.
FAU - Wang, B S
AU  - Wang BS
AD  - Institutes of Toxicology, College of Medicine, National Taiwan University, 
      Taipei, ROC.
FAU - Chen, Y J
AU  - Chen YJ
FAU - Liu, S H
AU  - Liu SH
FAU - Lin-Shiau, S Y
AU  - Lin-Shiau SY
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China (Republic : 1949- )
TA  - Proc Natl Sci Counc Repub China B
JT  - Proceedings of the National Science Council, Republic of China. Part B, Life 
      sciences
JID - 8502426
RN  - 0 (Anions)
RN  - 0 (Free Radicals)
RN  - 0 (Ion Channels)
RN  - EC 1.6.3.- (NADPH Oxidases)
RN  - Q1O6DSW23R (4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid/*pharmacology
MH  - Animals
MH  - *Anions
MH  - Ascitic Fluid/*cytology
MH  - Calcium/analysis
MH  - Enzyme Activation/drug effects
MH  - Free Radicals
MH  - Hydrogen-Ion Concentration
MH  - Ion Channels/*antagonists & inhibitors
MH  - Luminescent Measurements
MH  - Male
MH  - Membrane Potentials/drug effects
MH  - Mice
MH  - Mice, Inbred ICR
MH  - Mitochondria/physiology/ultrastructure
MH  - NADPH Oxidases/metabolism
MH  - Neutrophils/drug effects/*metabolism
EDAT- 2000/11/22 11:00
MHDA- 2001/03/03 10:01
CRDT- 2000/11/22 11:00
PHST- 2000/11/22 11:00 [pubmed]
PHST- 2001/03/03 10:01 [medline]
PHST- 2000/11/22 11:00 [entrez]
PST - ppublish
SO  - Proc Natl Sci Counc Repub China B. 2000 Oct;24(4):178-86.