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Oxidative stress: a key regulator of leiomyoma cell survival
Nicole M. Fletcher, Ph.D., Mohammed S. Abusamaan, M.D., Ira Memaj, B.S., Mohammed G. Saed, B.A., Ayman Al-Hendy, M.D., Ph.D., Michael P. Diamond, M.D., Ghassan M. Saed, Ph.D. Fertility and Sterility Volume 107, Issue 6, Pages e1 (June 2017) DOI: /j.fertnstert Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions
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Figure 1 Levels of myeloperoxidase (MPO) in myometrial and leiomyoma cells. Human myometrial and leiomyoma cells were treated with and without dichloroacetate (DCA) (20 μg/mL) and with and without hypoxia (2% O2, 5% CO2-nitrogen gas balance) for 24 hours. (A) Levels of MPO mRNA were determined with the use of real-time reverse-transcription polymerase chain reaction, and (B) levels of MPO protein were detected with the use of ELISA. Data were analyzed by means of unpaired t tests between groups and one-way analysis of variance to compare treatments within groups, with significant interactions further analyzed with the use of Tukey post hoc analysis with Bonferroni correction. All experiments were performed in triplicate. *P<.05 compared with untreated myometrial cells; **P<.05 compared with untreated leiomyoma cells; #P<.05 compared with hypoxia-treated myometrial cells; ##P<.05 compared with hypoxia-treated leiomyoma cells. Error bars represent SD of the mean. Fertility and Sterility , e1DOI: ( /j.fertnstert ) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions
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Figure 2 Levels of inducible nitric oxide synthase (iNOS) and nitrate/nitrite in myometrial and leiomyoma cells. Human myometrial cells and leiomyoma cells were treated with and without dichloroacetate (DCA) (20 ug/mL) and with and without hypoxia (2% O2, 5% CO2-nitrogen gas balance) for 24 hours. (A) Levels of iNOS mRNA were determined with the use of real-time reverse-transcription polymerase chain reaction, and (B) levels of total nitrate/nitrite, a surrogate of iNOS activity, were detected with the use of ELISA. Data were analyzed by means of unpaired t tests between groups and one-way analysis of variance to compare treatments within groups, with significant interactions further analyzed with the use of Tukey post hoc analysis with Bonferroni correction. All experiments were performed in triplicate. *P<.05 compared with untreated myometrial cells; **P<.05 compared with untreated leiomyoma cells; #P<.05 compared with hypoxia-treated myometrial cells; ##P<.05 compared with hypoxia-treated leiomyoma cells. Error bars represent SD of the mean. Fertility and Sterility , e1DOI: ( /j.fertnstert ) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions
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Figure 3 Apoptosis in myometrial and leiomyoma cells. Human myometrial cells and leiomyoma cells were treated with and without dichloroacetate (DCA) (20 μg/mL) and with and without hypoxia (2% O2) for 24 hours. (A) The ratio of antiapoptotic Bcl-2 to proapoptotic Bax was determined with the use of real-time reverse-transcription polymerase chain reaction, and (B) the level of caspase-3 activity was determined with the use of ELISA. For caspase-3 activity, the y-axis shows the absorbance at 405 nm, with increasing or decreasing absorbance corresponding to increasing or decreasing caspase-3 activity. All data were analyzed by means of unpaired t tests between groups and one-way analysis of variance to compare treatments within groups, with significant interactions further analyzed with the use of Tukey post hoc analysis with Bonferroni correction. All experiments were performed in triplicate. *P<.05 compared with untreated myometrial cells; **P<.05 compared with untreated leiomyoma cells; #P<.05 compared with hypoxia-treated myometrial cells; ##P<.05 compared with hypoxia-treated leiomyoma cells. Error bars represent SD of the mean. Fertility and Sterility , e1DOI: ( /j.fertnstert ) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions
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Figure 4 Summary of the role of oxidative stress in the pathogenesis of leiomyoma cells. BAX = Bcl-2–like protein 4; Bcl-2 = B-cell lymphoma 2; Cl− = chloride ion; DCA = dichloroacetate; H2O2 = hydrogen peroxide; H4B = tetrahydrobiopterin; HOCl = hypochlorous acid; iNOS = inducible nitric oxide synthase; L-Arg = L-arginine; MPO = myeloperoxidase; NO = nitric oxide; NO2− = nitrite; NO3− = nitrate; NO•+ = nitrosonium cation; O2•− = superoxide; ONOO− = peroxynitrite. Fertility and Sterility , e1DOI: ( /j.fertnstert ) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions
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