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(Hypertension. 2007;50:e167.)
© 2007 American Heart Association, Inc.
Letters to the Editor |
Department of Obstetrics and Gynecology, University of Debrecen Medical and Health Science Center, Debrecen, Hungary
cs
Department of Public Health, University of Debrecen Medical and Health Science Center, Debrecen, Hungary
Department of Obstetrics and Gynecology, University of Debrecen Medical and Health Science Center, Debrecen, Hungary
We read with interest the article by Tsukimori et al1 analyzing the relationship between the superoxide-anion (O2–) production of neutrophils during pregnancy in women with preeclampsia and essential hypertension. The results of the study demonstrated that n-formyl-methionyl-leucyl-phenylalanine stimulated O2– production of neutrophils during pregnancy was increased in women with preeclampsia compared with healthy pregnant women. There was no significant difference in the neutrophil O2– production in normal pregnancy between pregnant and postpartum women. In association with this, the authors previous study shows that there is no significant difference in O2– production in nonpregnant and healthy pregnant women.2
Several studies take up a question with the O2– production in pregnancy and preeclampsia, but the results are contradictory. Our unpublished results come to the conclusion that O2– production of neutrophils (stimulated by n-formyl-methionyl-leucyl-phenylalanine or phorbol-12.13-dibutyrate) in women with preeclampsia is significantly increased compared with healthy pregnant women; and in normal pregnant women, the granulocytes O2– production significantly decreases compared with nonpregnant and postpartum women concordant with the Crocker et al3 study presented earlier. Furthermore, in our examinations, there was no significant difference in O2– production between nonpregnant and preeclamptic pregnant women. The average O2– production of the granulocytes in nonpregnant women was 1.184 nmol/min per 3x105 cells, in healthy pregnant women was 0.472 nmol/min per 3x105 cells, and in preeclamptic patients was 1.106 nmol/min per 3x105 cells, stimulated by n-formyl-methionyl-leucyl-phenylalanine. These discrepancies give a chance for contradictory conclusions.
We assume that the cause of this deviation is the HEPES buffer, used by the authors for >1 hour at 37°C for incubating neutrophils. Incubation of cells in HEPES buffer for >60 minutes or less, depending on protocol conditions, could exposed cells to a nonphysiological surrounding toward the end of the incubation period. The nonphysiological environment includes the alkaline pH, toxic, and pharmacological effects as principals. HEPES interferes with the function of some ion channels, including calcium transport, which is important in O2– production.4,5 Contrarily, the Hanks buffer, used by us, is of a very simple composition and does not cause such a nonphysiological environment.5
We would like the authors of the statement to respond to our concerns. The number of studies in connection with O2– is increasing even in obstetrics. Minute differences in laboratory methods may result in contradictory conclusions. A standardized method might be helpful to solve this problem.
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2. Tsukimori K, Fukushima K, Tsushima A, Nakano H. Generation of reactive oxygen species by neutrophils and endothelial cell injury in normal and preeclamptic pregnancies. Hypertension. 2005; 46: 696–700.
3. Crocker IP, Wellings RP, Fletcher J, Baker PN. Neutrophil function in women with pre-eclampsia. Br J Obstet Gynaecol. 1999; 106: 822–288.[Medline] [Order article via Infotrieve]
4. Lelong IH, Rebel G. pH drift of "physiological buffers" and culture media used for cell incubation during in vitro studies. J Pharmacol Toxicol Methods. 1998; 39: 203–210.[CrossRef][Medline] [Order article via Infotrieve]
5. Lopez-Colome AM, Pasantes-Morales H. Effect of taurine on 45CA transport in frog retinal rod outer segments. Exp Eye Res. 1981; 32: 771–780.[CrossRef][Medline] [Order article via Infotrieve]
This article has been cited by other articles:
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K. Tsukimori, H. Nakano, and N. Wake Response to Preeclamptic Superoxide-Anion Production: Is There an Increase or a Failure of Reduction? Hypertension, November 1, 2007; 50(5): e168 - e164. [Full Text] [PDF] |
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