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Inhibition of nitric oxide: Effects of interleukin-1-beta-enhanced ovulation rate, steroid hormones and ovarian leukocyte distribution at ovulation in the rat


, : Inhibition of nitric oxide: Effects of interleukin-1-beta-enhanced ovulation rate, steroid hormones and ovarian leukocyte distribution at ovulation in the rat. Biology of Reproduction 54(2): 436-445

The ovulatory process resembles an inflammatory reaction with an infiltration of leukocytes, production of inflammatory mediators such as cytokines, and a general edema and hyperemia. Nitric oxide (NO), a potent vasodilator and the main mediator of macrophage tumoricidal and bacteriocidal activities, is known to participate in inflammatory reactions and has been shown to mediate the interleukin-1-beta (IL-1-beta)-directed tissue-remodeling events within the ovary. The regulation by NO of ovulation rate, leukocyte distribution, and steroid release in the rat ovary was investigated through use of a combination of in vivo and in vitro models of ovulation and a competitive inhibitor, N-omega-nitro-L-arginine methyl ester (L-NAME), of the NO synthase (NOS) enzyme. Subcutaneous L-NAME (1.5 times 10-4 mol/kg) administration significantly reduced the in vivo ovulation rate of eCG/hCG-primed rats (L-NAME-treated: 10.6 +- 1.8 (mean +- SEM) oocytes per ovary (O/O), 11.0 +- 1.2 rupture sites per ovary (RS/O; saline-treated: 18.0 +- 1.8 O/O, 19.4 +- 1.1 RS/O; p lt 0.01) at 20 h post-hCG. These results were reflected in vitro, where addition of L-NAME (3.5 times 10-5 mol/L) to LH (0.1 mu-g/ml)-perfused ovaries decreased ovulation rate from 8.2 +- 1.6 to 2.7 +- 1 ovulations per ovary (p lt 0.05) and simultaneously decreased nitrite accumulation at the completion of perfusions from 16.5 +- 1.9 to 4.1 +- 0.5 nmol/ml (p lt 0.001). The addition of L-NAME to LH + IL-1-beta (4 ng/ml)-perfused ovaries decreased ovulation rate from 15.2 +- 2.4 to 0.8 +- 0.8 ovulations per ovary (p lt 0.001) and simultaneously decreased nitrite accumulation at 22 h from 22.8 +- 2.2 to 1.9 +- 0.6 nmol/ml (p lt 0.001). Studies analyzing and manipulating perfusion flow rate indicated that the L-NAME effects on ovulation rate are primarily due to a reduction in flow rate resulting from inhibition of NO, which may be a consequence of the known vasoconstrictor effects of NOS inhibitors. The observed reduction of in vivo ovulation rate by NO inhibition at 20 h post-hCG was associated with a significant reduction in thecal MCA149+ neutrophils at 12 h post-hCG, the expected time of ovulation (L-NAME-treated: 98.4 +- 9.2 cells per thecal area; saline-treated: 211.5 +- 11.5 cells per thecal area; p lt 0.001), while ED1 + monocytes/macrophages underwent similar but nonsignificant changes. Plasma (20 h post-hCG) and perfusate progesterone were not different with L-NAME treatment, while perfusate estradiol levels were markedly reduced upon addition of L-NAME, suggesting a role for NO in ovulation but not in the process of luteinization. In summary, deprivation of NO by use of the competitive inhibitor, L-NAME, led to fewer ovulations, reduced accumulation of nitrite, a decreased neutrophil count in the theca of preovulatory follicles, and reduced estradiol secretion, while progesterone release remained unaffected. The NO pathway may therefore play an important role in the regulation of ovulation and the mediation of IL-1-beta's pro-ovulatory effects. There are likely to be primarily vascular effects, but also a nonvascular component, to the NO regulation of ovulation, with both components indirectly affecting ovulatory leukocyte distribution and steroid secretion.

US$29.90

PMID: 8788197


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