Application of H151 in preparation of product for improving ovulation function and oocyte quality

Abstract

The invention provides application of H151 in preparation of a product for improving an ovulation function and improving oocyte quality, and relates to the technical field of biomedicine.The H151 is supplemented to an old female mouse, it is found that the H151 can make the mood cycle of the old female mouse tend to be regularized, meanwhile, the ovulation number of the old female mouse can be remarkably increased, the number of abnormal oocytes can be reduced, and the ovulation function and the oocyte quality of the old female mouse can be improved. The ovulation function of female animals is obviously improved; meanwhile, H151 can reduce the number of spindle body morphological abnormalities of oocytes of the aged female rats, and H151 is beneficial to maintaining normal separation of chromosomes during subtraction second division of the oocytes, so that the phenomenon of chromosome dislocation during subtraction second division of the oocytes of the aged female rats is improved, and the probability of aneuploidy in the oocytes in the aging process is reduced. The invention provides powerful evidence for the H151 in preparation of products for improving ovulation function of aging female animals and improving oocyte quality.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to application of H151 in preparing products for improving ovulation function and enhancing oocyte quality. Background Art

[0002] Studies have shown that female fertility begins to decline around the age of 32, and the rate of decline accelerates after the age of 40, leading to an increase in oocyte aneuploidy, impaired early embryo development potential, and an increase in spontaneous abortion rates. Women are born with a certain number of oocytes, and this number and quality decrease significantly with age. In particular, when women reach the age of 35, the function of oocytes begins to decline significantly, which directly affects fertility. Although assisted reproductive technology can help infertile women to a certain extent, its treatment effect is limited, and egg quality is a key factor in determining the success rate of assisted reproductive technology. Therefore, there is an urgent need to explore methods to improve the ovarian ovulation function and egg quality of elderly women.

[0003] At present, researchers are trying to explore measures to prevent and treat ovarian aging in various directions. Existing reports mainly include the following aspects: drug intervention, stem cell therapy, mitochondrial transplantation, in vitro ovarian activation, ovarian transplantation, artificial 3D ovaries, etc. Looking at the above research directions, there are some problems, such as uncertain efficacy, ethical issues, immature technology, and low patient acceptance. On the other hand, drug treatment is conservative but safe and has high public acceptance. Among the drug methods, there are Chinese medicine health products. For example, the patent with publication number CN106473142A invented a health food that improves women's sleep and delays ovarian aging in middle-aged women. It also has a good alleviating effect on menopausal depression in middle-aged women, but this type of Chinese medicine health products is slow to take effect, has large individual differences, and has uncertain efficacy.

[0004] H151 is a potent, irreversible, and selective small molecule STING inhibitor that inhibits palmitoylation by covalently binding to the Cys91 site of STING, thereby effectively blocking the activity of STING. This mechanism enables H151 to significantly inhibit STING-mediated signaling, thereby alleviating the inflammatory response triggered by the STING / NF-κB pathway and reducing the release of proinflammatory cytokines, such as IL-6 and IL-23, which play an important role in the aging process. By inhibiting these inflammatory signals, H151 helps to reduce tissue damage and functional decline. Studies have shown that in the kidney and liver, various aging-related immune marker genes are significantly weakened after H151 inhibits STING. There are currently no reports of H151 improving age-related ovulation function. Summary of the invention

[0005] 1. Technical issues to be resolved

[0006] In view of the deficiencies of the prior art, the present invention provides the use of H151 in preparing products for improving ovulation function and increasing oocyte quality.

[0007] (II) Technical solution

[0008] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0009] In a first aspect, the present invention provides the use of H151 in preparing a product for improving the estrous cycle of aging animals.

[0010] In a second aspect, the present invention provides the use of H151 in preparing a product for improving the ovulation function of aged ovaries.

[0011] In a third aspect, the present invention provides the use of H151 in preparing a product for reducing the number of spindles with abnormal morphology in oocytes.

[0012] The above-mentioned products include medicines and health foods. The above-mentioned products contain an effective dose of H151, and the chemical formula of the H151 is shown in Formula I:

[0013]

[0014] The present invention does not specifically limit the source of the H151, and any H151 known in the art can be used. The product is preferably an injection.

[0015] The present invention has no special restrictions on the concentration of H151 in the product, and ensures that the dosage of H151 is not less than 750nM / d, and the recommended human supplement dosage is not less than 15μM / d.

[0016] (III) Beneficial effects

[0017] The present invention finds that by supplementing H151 to aged female mice, H151 can make the estrous cycle of aged female mice tend to be regular, and can significantly increase the number of ovulations of aged female mice, reduce the number of abnormal oocytes, and have a significant improvement effect on the ovulation function of female animals; at the same time, H151 can reduce the number of spindle morphology abnormalities in aged female mouse oocytes, and H151 is conducive to maintaining the normal separation of chromosomes during the second meiotic division of oocytes, improving the phenomenon of chromosome misalignment during the second meiotic division of aged female mouse oocytes, and reducing the probability of aneuploidy in oocytes during the aging process. The above results provide strong evidence for the use of H151 in the preparation of products that improve the ovulation function of aged female animals and improve the quality of oocytes. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Representative graphs of estrous cycle monitoring of some female mice in two groups.

[0019] Figure 2 The figures are the statistical results of estrous cycle changes of some female mice in two groups.

[0020] Figure 3 The following are comparison pictures of the microscopic morphology of mature oocytes of two groups of female mice.

[0021] Figure 4 The figures are the statistical results of the ovulation numbers of female mice in the two groups; compared with DMSO, *p<0.05, **p<0.01.

[0022] Figure 5 The results are the ratios of the number of abnormal morphological oocytes / normal morphological oocytes in the three groups of female mice; compared with DMSO, *p<0.05, **p<0.01.

[0023] Figure 6 Comparison of the microscopic morphology of spindles and chromosomes in oocytes of two groups of female mice.

[0024] Figure 7 The statistical results of spindle abnormality rates of oocytes of female mice in the two groups; compared with DMSO, *p<0.05, **p<0.01. DETAILED DESCRIPTION

[0025] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Example

[0027] 1. Effects of H151 on the estrous cycle of aged female mice

[0028] Twenty healthy C57 / BL6J female mice aged 8 months, weighing (22±2)g, were randomly divided into an experimental group (H151) and a control group (DMSO), with 10 mice in each group. The female mice in the experimental group were intraperitoneally injected with H151 at 750nM / mouse / d; the female mice in the control group were gavaged with an equal amount of solvent, and the solvent ratio was 5% Tween-80+5% DMSO+90% PBS. After the two groups of female mice were treated for 8 weeks, follow-up experiments were performed.

[0029] Vaginal exfoliated cell smears were collected every morning for 14 consecutive days for hematoxylin and eosin staining and observation. The judgment criteria were:

[0030] Proestrus: numerous round nucleated cells, uniform in size and appearance;

[0031] Estrus: a large number of anucleated keratinized epithelial cells;

[0032] Metestrus: numerous leukocytes and a few large, agranular, anucleated keratinized epithelial cells;

[0033] Diestrus: prominent polymorphonuclear leukocytes and a few epithelial cells and keratinocytes.

[0034] To determine the estrous cycle stage. Figure 1 .

[0035] from Figure 2 It can be seen that compared with the H151 group, the estrous cycle of the DMSO group was disordered, partially manifested as long estrus and long diestrus, while the estrous cycle of the H151 group was relatively regular, without long estrus and long diestrus.

[0036] 2. Effects of H151 on ovulation function in aged female mice

[0037] The above-mentioned experimental group (H151) and control group (DMSO) female mice were treated for ovulation induction. Each mouse in each group was injected with 10 IU of pregnant mare serum gonadotropin (PMSG), and 48 hours later, 10 IU of human chorionic gonadotropin (hCG) was injected. 14 hours later, the cumulus oocyte complexes (COCs) discharged into the ampulla of the fallopian tube were removed, and then 0.1% hyaluronidase was used to remove the granulosa cells around the oocytes. The morphology of the oocytes was observed and counted under a Zeiss laser confocal microscope LSM880.

[0038] The results are as follows Figure 3-5 As shown, compared with the control group, the oocytes of the experimental group were more regular, round and full, the fragmentation of oocytes was significantly reduced, the number of ovulations was significantly increased (P<0.05), and the abnormal rate of oocytes was significantly reduced (P<0.05). The above results show that H151 can significantly increase the number of ovulations and the quality of oocytes in elderly female animals, and has a significant effect on improving the ovulation function of female animals.

[0039] 3. Effects of H151 on the spindle of oocytes in aged female mice

[0040] The above-mentioned experimental group (H151) and control group (DMSO) female mice were treated for ovulation induction. Each mouse was injected with 10 IU of pregnant mare serum gonadotropin (PMSG), and 48 hours later, 10 IU of human chorionic gonadotropin (hCG) was injected. 14 hours later, the cumulus oocyte complexes (COCs) discharged into the ampulla of the fallopian tube were removed, and the granulosa cells around the oocytes were removed with 0.1% hyaluronidase. The obtained oocytes were fixed in paraformaldehyde for 30 minutes, permeabilized with 0.5% TritonX-100 for 20 minutes, blocked with 1% BSA for 1 hour, and incubated with 1:200 anti-α-tubulinmonoclonal antibody at 4°C overnight. After washing three times with IVF solution, the cells were incubated with FITC-conjugated secondary antibody at room temperature for 1 h, and then the nuclei were stained with propidium iodide (PI) and transferred into 10 μl antifade droplets. The spindle morphology and chromosome arrangement were observed under a Zeiss laser confocal microscope LSM880.

[0041] The results are as follows Figure 6-7 As shown, compared with the control group, the spindle morphology of the oocytes of the female mice in the experimental group was regular and without obvious abnormalities. The abnormality rate of the oocytes of the female mice in this group was significantly lower than that in the control group (P<0.05). The above results showed that H151 can significantly reduce the abnormality of the oocyte spindles in aged female animals, which is beneficial to maintain the normal separation of chromosomes during the second meiotic division of the oocytes, improve the phenomenon of chromosome misalignment during the second meiotic division of the oocytes of aged female mice, and reduce the probability of aneuploidy in oocytes during the aging process.

[0042] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Application of H151 in the preparation of products for improving the estrous cycle of aging animals.

2. Application of H151 in the preparation of products for improving the ovulation function of aging ovaries.

3. Application of H151 in the preparation of products for reducing the number of abnormal spindles in oocytes.

4. The product according to any one of claims 1 to 3, characterized in that: The products include medicines and health foods.

5. The product according to any one of claims 1 to 3, characterized in that: The product contains an effective dose of H151, and the chemical formula of H151 is shown in Formula I:

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