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, 13 (4), 307-314

Ameliorative Effect of Crocin on Sperm Parameters and In Vitro Fertilization in Mice Under Oxidative Stress Induced by Paraquat

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Ameliorative Effect of Crocin on Sperm Parameters and In Vitro Fertilization in Mice Under Oxidative Stress Induced by Paraquat

Fahime Sadat Kamali et al. Int J Fertil Steril.

Abstract

Background: Paraquat (PQ) is an herbicide that is genotoxic and cytotoxic for male germ cells. In this study, we investigated the protective role of crocin (Cr) against the destructive effects of PQ on sperm quality and in vitro fertilization (IVF) outcomes.

Materials and methods: In this experimental study, a total of 28 male mice (20-25 g) were divided into four groups: control, which received intraperitoneal (IP) injections of 0.1 ml normal saline per day; PQ group received IP injections of PQ (5 mg/kg/day); experimental (PQ+Cr group) received PQ along with IP injections of Cr (200 mg/kg/day); and positive control (Cr) received IP injections of Cr (200 mg/kg/day). In the last two weeks of the treatment period (35 days of treatment), 16 non-pregnant mice were stimulated to receive adult oocytes. At the end of the treatment period, after euthanizing the mice, the sperms were extracted from the epididymis of each mouse and prepared for evaluation of sperm parameters and IVF.

Results: In the PQ+Cr group, Cr caused a significant increase in the average number of sperms and the mean percentage of motile and viable sperms. There was a significant decrease in the mean number of immature and DNA-damaged sperms compared to the PQ group (P<0.001). IVF evaluation in the PQ+Cr group showed that the mean percentage of fertilization, two- and four-cell embryos, blastocysts, and hatched embryos significantly increased. Cr caused a significant decrease in the mean percentage of the arrested embryos compared to the PQ group (P<0.001). However, the Cr group did not have any toxic effects on sperm quality or IVF results.

Conclusion: The findings of this study showed that Cr, due to its effective and potent antioxidant properties, could reduce or suppress the destructive effects on sperm parameters and IVF caused by PQ.

Keywords: Crocin; In vitro Fertilization; Mice; Paraquat; Sperm.

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Fig 1
Fig 1
Dissected fallopian tube for obtaining oocyte mass. A. Fallopian tubes (1), Ampulla (2) and B. Ampulla (1), Detached oocyte masses (2) (magnification: ×40).
Fig 2
Fig 2
The dissecting steps.
Fig 1
Fig 1
Evaluation of sperm viability, maturity and DNA damage. A. Mature sperm are pale (1) immature sperm are light to dark and blue colour (2) (aniline blue, magnification: ×400), B. Nonviable sperm are pink to red colour (1) and the viable sperm are achromatic (2) (eosin-nigrosin, magnification: ×400), and C. Sperm without (1) and with (2) DNA damage [Acridine orange (AO), ×400 magnification considering the camera magnification of ×300].
Fig 4
Fig 4
Zygotes and early embryos in different stages of development or arresting. A. In the control group, several embryos are seen in the blastocyst and hatching stages, B. In the PQ group, several hatched embryos are present at the blastocyst stage, and numerous infertile oocytes and arrested embryos are present and C. In the paraquat plus crocin (PQ+Cr) group, several hatched embryos are present in the blastocyst stage. There are several arrested embryos present, (magnification: ×300). 1; Blastocyst, 2; Hatched embryo, and 3; Arrested embryo.

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