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Application of melatonin in preparation of medicine for improving oocyte quality

A technology of oocytes and melatonin, which is applied in the field of biomedicine, can solve the problems affecting the maturation of oocytes and the decrease of the blastocyst formation rate in mice, and achieve the effect of improving reproductive ability

Inactive Publication Date: 2020-02-25
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Kendall V et al. found that when 8mM cotinine was added to the early embryo culture medium of mice, it could lead to a significant decrease in the rate of blastocyst formation in mice
However, whether cotinine affects the maturation of oocytes has not yet been reported.
[0004] At present, there is no research on the correlation between cotinine and oocyte quality in the prior art. If a method can be developed to alleviate the effect of cotinine on oocyte quality, it will be effective in reducing the risk of female infertility caused by smoking. Treatment has important theoretical guiding significance

Method used

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  • Application of melatonin in preparation of medicine for improving oocyte quality
  • Application of melatonin in preparation of medicine for improving oocyte quality
  • Application of melatonin in preparation of medicine for improving oocyte quality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Melatonin relieves cotinine-induced decline in oocyte quality

[0043]Prepare oocyte maturation medium containing 800 μmol / L cotinine and 0 μmol / L, 0.1 μmol / L, 5 μmol / L, 10 μmol / L, 15 μmol / L, 20 μmol / L melatonin in advance. Make a 60 μL oocyte culture drop in a 60 mm Petri dish at 37 °C, 5% CO 2 Pre-equilibrate for 2 hours. Put 30-35 collected GV stage oocytes into each droplet and place them in an incubator for culture. After culturing for 16 hours, the oocyte maturation rate, that is, the first polar body discharge rate, was counted. The result is as figure 1 Shown: The addition of melatonin can significantly alleviate the effect of cotinine on oocyte first polar body excretion. The addition of 0.1μmol / L, 5μmol / L, 10μmol / L, 15μmol / L, 20μmol / L MT can alleviate the effect of cotinine on the first polar body excretion of oocytes.

[0044] Collect the above-mentioned oocytes (MII stage) that have discharged the first polar body, and transfer them to 2% paraformaldehy...

Embodiment 2

[0047] Melatonin can improve the fertilization rate of MII stage oocytes

[0048] In order to visually evaluate the quality of MII stage oocytes, MII stage oocytes were parthenogenetically activated and the pronuclear rate was detected.

[0049] The specific method is as follows:

[0050] (1) Prepare parthenogenetic activation solutions A (in vitro fertilization fluid containing 100 μM strontium chloride and 5 mg / L cytochalasin D) and B (in vitro fertilization fluid containing 5 mg / L cytochalasin D), and then place them in a 60 mm culture dish Make a 60 μL fertilization droplet at 37°C, 5% CO 2 Medium pre-balance 8-12h.

[0051] (2) Put MII stage oocytes into parthenogenetic activation solution A for 2.5 hours, then put them into parthenogenetic activation solution B for 3.5 hours, and then count the pronuclear rate. The result is as Image 6 As shown, melatonin can significantly alleviate the low rate of pronuclei induced by cotinine, that is, increase the rate of pronucl...

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Abstract

The invention discloses an application of melatonin in preparation of a medicine for improving oocyte quality. According to the invention, melatonin is directly added into an oocyte in-vitro maturation culture solution, which proves that the melatonin can relieve the reduction of the quality of the cotinine-induced oocytes and is shown as reduction of the ROS level in the oocytes, reduction of aneuploidy formation, promotion of uniform distribution of actin, improvement of the form of spindles and improvement of mitochondrial functions, and then the later fertilization rate of the oocytes is remarkably increased. The invention provides a method for improving the influence of the main metabolite cotinine of nicotine in the blood of a smoker on the quality of female oocytes, and provides aneffective method for improving the reproductive capacity of smoking women.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of melatonin in the preparation of medicines for improving the quality of oocytes. Background technique [0002] Melatonin (Melatonin, MT) is a highly lipophilic and partially hydrophilic neuroendocrine hormone secreted by the pineal gland, and its secretion has a clear rhythm of low day and night high. In recent years, melatonin has been widely used and paid attention to. There are researches on its antioxidant effect at home and abroad, and it has been shown that it is a powerful free radical scavenger of reactive oxygen species (ROS) and reactive nitrogen species (RNS). agent to protect organs and cells from damage. Melatonin can improve the impact of harmful substances on the quality of oocytes has been related research. [0003] Although tobacco is the most avoidable cause of death, approximately 5 million people die each year from tobacco-r...

Claims

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Application Information

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IPC IPC(8): A61K31/4045A61P15/08
CPCA61K31/4045A61P15/08
Inventor 程金妹孙斐米盼盼
Owner NANTONG UNIVERSITY
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