褪黑素可调节BCL-xl的表达，并改善通过体外受精获得的玻璃化小鼠胚胎的发育情况。
Melatonin modulates the expression of BCL-xl and improve the development of vitrified embryos obtained by IVF in mice.
作者信息
Dehghani-Mohammadabadi Maryam, Salehi Mohammad, Farifteh Fattaneh, Nematollahi Sedigheh, Arefian Ehsan, Hajjarizadeh Atena, Parivar Kazem, Nourmohammadi Zahra
机构信息
Department of Transgenic Animal Science, Stem Cell Technology Research Center, Tehran, Iran.
出版信息
J Assist Reprod Genet. 2014 Apr;31(4):453-61. doi: 10.1007/s10815-014-0172-9. Epub 2014 Jan 14.
PURPOSE
Antioxidant and anti-apoptotic effects of melatonin on development of in vitro fertilization (IVF)/vitrified two-cell mouse embryos were evaluated in this study.
METHODS
The IVF two-cell embryos were vitrified by cryotop, and were cultured in KSOM medium in different concentrations of melatonin (10(-6), 10(-9), 10(-12) M) and without melatonin. The blastocyst cell number, apoptotic cells and glutathione (GSH) level were evaluated by differential, TUNEL and cell tracker blue staining, respectively. The expression of Bax and Bcl-xl genes was evaluated by qPCR. The expression of melatonin receptors (Mtnr1a and Mtnr1b) in mouse 2-cell embryos and blastocysts was evaluated by RT-PCR.
RESULTS
Melatonin increased the rate of cleavage and blastulation at 10(-12) M concentration (p < 0.05). The number of trophectoderm and inner cell mass showed a significant increase (p < 0.05) in 10(-9) M melatonin. The 10(-9) M and 10(-12) M melatonin treatments significantly reduced (p < 0.05) the apoptotic index. The significant increase in the expression of Bcl-xl observed at 10(-9) M concentration however, reduced expression of Bax was not statistically significant. The levels of GSH in 10(-9) and 10(-12) M groups were significantly improved relative to the control group (p < 0.05). The Mtnr1a was expressed in 2-cell embryos and blastocysts in all groups, but the expression of Mntr1b was not detected.
CONCLUSION
Melatonin may have a special role against oxidative stress in protection of IVF/vitrified embryos.
目的
本研究评估褪黑素对体外受精（IVF）/玻璃化二细胞小鼠胚胎发育的抗氧化和抗凋亡作用。
方法
将IVF二细胞胚胎用冷冻环进行玻璃化处理，并在含有不同浓度褪黑素（10⁻⁶、10⁻⁹、10⁻¹² M）和不含褪黑素的KSOM培养基中培养。分别通过微分染色、TUNEL染色和细胞追踪蓝染色评估囊胚细胞数量、凋亡细胞和谷胱甘肽（GSH）水平。通过qPCR评估Bax和Bcl-xl基因的表达。通过RT-PCR评估小鼠二细胞胚胎和囊胚中褪黑素受体（Mtnr1a和Mtnr1b）的表达。
结果
褪黑素在10⁻¹² M浓度时可提高卵裂率和囊胚形成率（p < 0.05）。在10⁻⁹ M褪黑素组中，滋养外胚层和内细胞团数量显著增加（p < 0.05）。10⁻⁹ M和10⁻¹² M褪黑素处理显著降低了凋亡指数（p < 0.05）。在10⁻⁹ M浓度时观察到Bcl-xl表达显著增加，然而Bax表达降低无统计学意义。相对于对照组，10⁻⁹ M和10⁻¹² M组的GSH水平显著提高（p < 0.05）。所有组的二细胞胚胎和囊胚中均表达Mtnr1a，但未检测到Mntr1b的表达。
结论
褪黑素在保护IVF/玻璃化胚胎免受氧化应激方面可能具有特殊作用。
Zotero 插件