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Application of 4,4'-dimethoxy chalcone in delaying of in-vivo and in-vitro aging of oocyte

A technology of dimethoxychalcone and oocytes, applied in the field of biomedicine, can solve the problems of anti-aging functions and mechanisms that are still blank

Active Publication Date: 2021-10-29
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on the anti-aging function and mechanism of 4,4'-dimethoxychalcone in mammalian reproductive system is still blank

Method used

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  • Application of 4,4'-dimethoxy chalcone in delaying of in-vivo and in-vitro aging of oocyte
  • Application of 4,4'-dimethoxy chalcone in delaying of in-vivo and in-vitro aging of oocyte
  • Application of 4,4'-dimethoxy chalcone in delaying of in-vivo and in-vitro aging of oocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 screened the optimal effective concentration of DMC

[0028] We chose the level of ROS, a classic indicator of oocyte aging after ovulation, as the criterion for judging the occurrence of aging. We simulated the aging process of MII oocytes in vitro by culturing for 12 hours, and set different concentration gradients of DMC from low to high (DMC dissolved in DMSO): 1, 10, 50, 100, 1000 μM were added to M2 medium (purchased from Sigma, product number M7167, the same below), and the fresh (Fresh) group and the aged (POA) group were set as controls. The same amount of DMSO was used to dissolve DMC in the administration group. The results show that the accumulation of ROS can be effectively inhibited when 10-1000 μM DMC is added to the culture system. In order to reduce the toxic and side effects of the drug on cells, we choose the lowest effective concentration of 10 μM ( figure 2 A, 2B).

Embodiment 2D

[0029] Example 2DMC reduces ROS levels and inhibits apoptosis in aged oocytes after ovulation in mice

[0030] After adding 10 μM DMC to the culture medium to culture the MII stage oocytes for 12 hours in vitro, collect the aged oocytes in vitro, put them into the oxidation-sensitive fluorescent probe DCFH-DA (1:250), and incubate them in a 37°C 5% CO2 incubator for 30 minutes. , observed under a fluorescence inverted microscope and found that DMC addition can effectively reduce the accumulation of ROS in aged MII oocytes in vitro ( image 3 A, 3B), high levels of ROS can induce the occurrence of early apoptosis. The apoptosis level of aged MII oocytes in vitro was detected by AnnexinV-FITC. The results showed that the apoptosis level of oocytes in the DMC addition group was significantly reduce( image 3 C, 3D).

Embodiment 3D

[0031] Example 3 DMC can improve the distribution of mitochondria in aged oocytes after ovulation in mice

[0032] MII stage oocytes were cultured in vitro by adding 10 μM DMC to M2 medium for 12 hours, then the in vitro aged oocytes were collected, placed in MitoTracker (1:5000) staining solution, and incubated at 37°C in a 5% CO2 incubator for 1 hour. The mitochondrial distribution was observed under a focusing microscope, and the results showed that the probability of abnormal mitochondrial distribution decreased after DMC was added ( Figure 4 A, 4B).

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Abstract

The invention discloses an application of 4,4'-dimethoxy chalcone in delaying of in-vivo and in-vitro aging of an oocyte. The invention discloses an application of the 4,4'-dimethoxy chalcone in preparation of a reagent for delaying an in-vitro aging process of the oocyte. The invention discloses an application of the 4,4'-dimethoxy chalcone in preparation of a medicine for improving reproductive aging. Results of the invention show that the DMC can be used for effectively improving the quality of in-vitro-aged MII-stage oocytes, and in-vitro fertilization experiments verify that the fertilization rate and blastocyst development rate of the aged MII-stage oocytes can be effectively improved by adding the DMC into a culture medium. By adding the DMC into a culture solution for in-vitro-aged MII oocytes of a mouse, the in-vitro aging process of the MII-stage oocytes can be effectively delayed, the oocyte quality is improved, and the ovulation rate can be effectively increased when an elderly mouse orally takes the DMC.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the application of 4,4'-dimethoxychalcone in delaying oocyte aging in vitro and in vivo. Background technique [0002] The quality of oocytes is affected by many factors, including lifestyle, environment and age, among which the decline in oocyte quality caused by age is the most important and irresistible factor. Driven by the social background of women marrying later and having children later, the impact of age-related fertility problems is becoming more and more common. It is well known that female fertility declines significantly with age, with a sharp decline in fertility around the age of 35. This age-related female infertility is characterized by a gradual decline in ovarian function, which is associated with a decrease in the number and quality of oocytes in the ovary. decline is closely related. The decline of egg quality in elderly women not only leads to an increase in the i...

Claims

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Application Information

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IPC IPC(8): C12N5/075A61K31/12A61P15/08A61K36/232
CPCC12N5/0609A61K31/12A61P15/08A61K36/232C12N2500/30
Inventor 霍然李明芮刘露黄婕何安澜
Owner NANJING MEDICAL UNIV
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