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The regulation of nitric oxide synthase isoform expression in mouse and human fallopian tubes: potential insights for ectopic pregnancy.

Hu J, Ma S, Zou S, Li X, Cui P, Weijdegård B, Wu G, Shao R, Billig H, Feng Y - Int J Mol Sci (2014)

Bottom Line: Nitric oxide (NO) is highly unstable and has a half-life of seconds in buffer solutions.It is synthesized by NO-synthase (NOS), which has been found to exist in the following three isoforms: neuro nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS).Elevated iNOS activity might influence ovulation, cilia beats, contractility, and embryo transportation in such a manner as to increase the risk of ectopic pregnancy.

View Article: PubMed Central - PubMed

Affiliation: Department of Integrative Medicine and Neurobiology, State Key Lab of Medical Neurobiology, Shanghai Medical College and Institute of Acupuncture Research (WHO Collaborating Center for Traditional Medicine), Institute of Brain Science, Fudan University, Shanghai 200032, China. 11211250005@fudan.edu.cn.

ABSTRACT
Nitric oxide (NO) is highly unstable and has a half-life of seconds in buffer solutions. It is synthesized by NO-synthase (NOS), which has been found to exist in the following three isoforms: neuro nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS). NOS activity is localized in the reproductive tracts of many species, although direct evidence for NOS isoforms in the Fallopian tubes of mice is still lacking. In the present study, we investigated the expression and regulation of NOS isoforms in the mouse and human Fallopian tubes during the estrous and menstrual cycles, respectively. We also measured isoform expression in humans with ectopic pregnancy and in mice treated with lipopolysaccharide (LPS). Our results confirmed the presence of different NOS isoforms in the mouse and human Fallopian tubes during different stages of the estrous and menstrual cycles and showed that iNOS expression increased in the Fallopian tubes of women with ectopic pregnancy and in LPS-treated mice. Elevated iNOS activity might influence ovulation, cilia beats, contractility, and embryo transportation in such a manner as to increase the risk of ectopic pregnancy. This study has provided morphological and molecular evidence that NOS isoforms are present and active in the human and mouse Fallopian tubes and suggests that iNOS might play an important role in both the reproductive cycle and infection-induced ectopic pregnancies.

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Time-dependent regulation of iNOS and eNOS expression in mouse Fallopian tubes following steroid hormone treatment. (A) The iNOS mRNA level in mouse Fallopian tubes in vitro. The expression of iNOS mRNA was significantly up-regulated from 6 to 24 h following P4 treatment but not following E2 or E2 plus P4 together (p < 0.05); (B) The eNOS mRNA level in mouse Fallopian tubes in vitro. The expression of eNOS mRNA was down-regulated from 6 to 24 h following E2 and E2 plus P4 treatment together but not after P4 treatment; (C) The nitric oxide concentration in tubal culture medium in vitro. The nitric oxide concentration was significantly lower after 6 h in tissue cultured with P4 or E2 plus P4. The nitrate concentration was lower after 24 h in culture medium containing E2 and E2 plus P4; and (D) The LDH level in the tubal culture medium as measured by the optical density on a microplate fluorometer at a wavelength of 490 nm. There was no significant difference between the four groups. Significance was tested by one-way ANOVA or two-way ANOVA. Values are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with 6 h incubation of same steroid hormone treatment; #p < 0.05, ##p < 0.01, ###p < 0.001 compared with vehicle at the same incubation time.
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ijms-16-00049-f005: Time-dependent regulation of iNOS and eNOS expression in mouse Fallopian tubes following steroid hormone treatment. (A) The iNOS mRNA level in mouse Fallopian tubes in vitro. The expression of iNOS mRNA was significantly up-regulated from 6 to 24 h following P4 treatment but not following E2 or E2 plus P4 together (p < 0.05); (B) The eNOS mRNA level in mouse Fallopian tubes in vitro. The expression of eNOS mRNA was down-regulated from 6 to 24 h following E2 and E2 plus P4 treatment together but not after P4 treatment; (C) The nitric oxide concentration in tubal culture medium in vitro. The nitric oxide concentration was significantly lower after 6 h in tissue cultured with P4 or E2 plus P4. The nitrate concentration was lower after 24 h in culture medium containing E2 and E2 plus P4; and (D) The LDH level in the tubal culture medium as measured by the optical density on a microplate fluorometer at a wavelength of 490 nm. There was no significant difference between the four groups. Significance was tested by one-way ANOVA or two-way ANOVA. Values are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with 6 h incubation of same steroid hormone treatment; #p < 0.05, ##p < 0.01, ###p < 0.001 compared with vehicle at the same incubation time.

Mentions: To determine whether ovarian steroid hormones regulate NOS expression in mouse Fallopian tubes, qRT-PCR was performed in mouse Fallopian tubes treated with E2 or P4 in vitro. The expression of iNOS mRNA was significantly up-regulated from 24 h following P4 treatment but not following E2 or a combination of E2 and P4 (p < 0.05), but compared with vehicle, iNOS mRNA level was decreased with 6 h P4 treatment (p < 0.05) (Figure 5A). In contrast, the expression of eNOS mRNA was down-regulated from 6 to 24 h following treatment with E2 (p < 0.05) and the combination of E2 and P4 (p < 0.001), but not P4. In addition, eNOS mRNA level was decreased with 24 h E2 (p < 0.05) and E2 plus P4 (p < 0.001) treatment compared with vehicle (Figure 5B).


The regulation of nitric oxide synthase isoform expression in mouse and human fallopian tubes: potential insights for ectopic pregnancy.

Hu J, Ma S, Zou S, Li X, Cui P, Weijdegård B, Wu G, Shao R, Billig H, Feng Y - Int J Mol Sci (2014)

Time-dependent regulation of iNOS and eNOS expression in mouse Fallopian tubes following steroid hormone treatment. (A) The iNOS mRNA level in mouse Fallopian tubes in vitro. The expression of iNOS mRNA was significantly up-regulated from 6 to 24 h following P4 treatment but not following E2 or E2 plus P4 together (p < 0.05); (B) The eNOS mRNA level in mouse Fallopian tubes in vitro. The expression of eNOS mRNA was down-regulated from 6 to 24 h following E2 and E2 plus P4 treatment together but not after P4 treatment; (C) The nitric oxide concentration in tubal culture medium in vitro. The nitric oxide concentration was significantly lower after 6 h in tissue cultured with P4 or E2 plus P4. The nitrate concentration was lower after 24 h in culture medium containing E2 and E2 plus P4; and (D) The LDH level in the tubal culture medium as measured by the optical density on a microplate fluorometer at a wavelength of 490 nm. There was no significant difference between the four groups. Significance was tested by one-way ANOVA or two-way ANOVA. Values are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with 6 h incubation of same steroid hormone treatment; #p < 0.05, ##p < 0.01, ###p < 0.001 compared with vehicle at the same incubation time.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4307235&req=5

ijms-16-00049-f005: Time-dependent regulation of iNOS and eNOS expression in mouse Fallopian tubes following steroid hormone treatment. (A) The iNOS mRNA level in mouse Fallopian tubes in vitro. The expression of iNOS mRNA was significantly up-regulated from 6 to 24 h following P4 treatment but not following E2 or E2 plus P4 together (p < 0.05); (B) The eNOS mRNA level in mouse Fallopian tubes in vitro. The expression of eNOS mRNA was down-regulated from 6 to 24 h following E2 and E2 plus P4 treatment together but not after P4 treatment; (C) The nitric oxide concentration in tubal culture medium in vitro. The nitric oxide concentration was significantly lower after 6 h in tissue cultured with P4 or E2 plus P4. The nitrate concentration was lower after 24 h in culture medium containing E2 and E2 plus P4; and (D) The LDH level in the tubal culture medium as measured by the optical density on a microplate fluorometer at a wavelength of 490 nm. There was no significant difference between the four groups. Significance was tested by one-way ANOVA or two-way ANOVA. Values are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with 6 h incubation of same steroid hormone treatment; #p < 0.05, ##p < 0.01, ###p < 0.001 compared with vehicle at the same incubation time.
Mentions: To determine whether ovarian steroid hormones regulate NOS expression in mouse Fallopian tubes, qRT-PCR was performed in mouse Fallopian tubes treated with E2 or P4 in vitro. The expression of iNOS mRNA was significantly up-regulated from 24 h following P4 treatment but not following E2 or a combination of E2 and P4 (p < 0.05), but compared with vehicle, iNOS mRNA level was decreased with 6 h P4 treatment (p < 0.05) (Figure 5A). In contrast, the expression of eNOS mRNA was down-regulated from 6 to 24 h following treatment with E2 (p < 0.05) and the combination of E2 and P4 (p < 0.001), but not P4. In addition, eNOS mRNA level was decreased with 24 h E2 (p < 0.05) and E2 plus P4 (p < 0.001) treatment compared with vehicle (Figure 5B).

Bottom Line: Nitric oxide (NO) is highly unstable and has a half-life of seconds in buffer solutions.It is synthesized by NO-synthase (NOS), which has been found to exist in the following three isoforms: neuro nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS).Elevated iNOS activity might influence ovulation, cilia beats, contractility, and embryo transportation in such a manner as to increase the risk of ectopic pregnancy.

View Article: PubMed Central - PubMed

Affiliation: Department of Integrative Medicine and Neurobiology, State Key Lab of Medical Neurobiology, Shanghai Medical College and Institute of Acupuncture Research (WHO Collaborating Center for Traditional Medicine), Institute of Brain Science, Fudan University, Shanghai 200032, China. 11211250005@fudan.edu.cn.

ABSTRACT
Nitric oxide (NO) is highly unstable and has a half-life of seconds in buffer solutions. It is synthesized by NO-synthase (NOS), which has been found to exist in the following three isoforms: neuro nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS). NOS activity is localized in the reproductive tracts of many species, although direct evidence for NOS isoforms in the Fallopian tubes of mice is still lacking. In the present study, we investigated the expression and regulation of NOS isoforms in the mouse and human Fallopian tubes during the estrous and menstrual cycles, respectively. We also measured isoform expression in humans with ectopic pregnancy and in mice treated with lipopolysaccharide (LPS). Our results confirmed the presence of different NOS isoforms in the mouse and human Fallopian tubes during different stages of the estrous and menstrual cycles and showed that iNOS expression increased in the Fallopian tubes of women with ectopic pregnancy and in LPS-treated mice. Elevated iNOS activity might influence ovulation, cilia beats, contractility, and embryo transportation in such a manner as to increase the risk of ectopic pregnancy. This study has provided morphological and molecular evidence that NOS isoforms are present and active in the human and mouse Fallopian tubes and suggests that iNOS might play an important role in both the reproductive cycle and infection-induced ectopic pregnancies.

Show MeSH
Related in: MedlinePlus