Application of ergothioneine in improving fertility of high-fertility-age female individuals

Abstract

The invention provides application of ergothioneine in improving fertility of high-fertility-age female individuals, and belongs to the technical field of natural medicines. The invention provides an application of EGT in preparation of a product for improving fertility of a high-fertility-age female individual. In the embodiment of the invention, a high-fertility-age ICR female mouse is taken as an animal model, the influence of the EGT on the fertility of the high-fertility-age female mouse is verified from in-vivo and in-vitro aspects, and the result shows that the EGT has the effect of improving the in-vitro fertilization rate and blastocyst rate of oocytes of the high-fertility-age female mouse; the superovulation number and the litter number of high-childbearing-age female mice are increased. It is proved that the EGT has the effect of improving the fertility of high-fertility-age female individuals.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of natural medicines, and particularly relates to application of ergothioneine in improving the fertility of high-age female individuals. BACKGROUND

[0002] The fertility of human beings will decrease sharply with the increase of age. The risk of infertility, spontaneous abortion, perinatal mortality and congenital abnormality of high-age women will increase significantly. Although assisted reproductive technology can make up for the suboptimal reproductive results of human beings to a certain extent, the success rate thereof will decrease with the increase of the age of women of childbearing age.

[0003] The main reasons for affecting the fertility of women of high age include the decrease of ovarian reserve function, the decrease of oocyte quality and the decrease of oocyte / embryo capacity, etc., which finally reduces the oocyte fertilization, implantation and development capacity, increases the risk of spontaneous abortion, fetal malformation, premature birth and stillbirth. Therefore, exploring and finding a method for improving the fertilization and development capacity of high-age oocytes can provide a feasible way for improving the fertility of women of high age, and has important scientific significance and research value.

[0004] Ergothioneine (EGT) is a safe, orally administered, natural hydrophilic amino acid. Due to its powerful antioxidant capacity, significant anti-aging properties, good safety profile, and easy absorption, it has been widely used in various commercially available skincare and health supplements. Currently, there are 10 clinical studies worldwide based on EGT, covering multiple areas including kidney failure, cognitive impairment, sleep, and immune regulation. Appropriate EGT supplementation can reduce the risk of chronic diseases and premature aging; therefore, EGT is also defined as a "longevity vitamin." Compared to traditional antioxidants (such as glutathione and vitamin E), EGT exhibits stronger free radical scavenging activity and can protect different subcellular organelles in multiple ways: When EGT enters the cell, it increases the catalytic activity of cGPDH by thiolation modification, while regulating the activity of PARPs and SIRTs family enzymes, indirectly affecting the regeneration, transport, and circulation of NAD+ in the cytoplasm and mitochondria, and directly affecting the level of ROS in mitochondria; on the other hand, it activates the Nrf2 / ARE pathway and corresponding antioxidant stress factors, including heme oxygenase-1 (HO-1), NADP(H)quinone oxidoreductase (NQO-1), and γ-glutamylcysteine ​​synthase (γ-GCLC), improving the intracellular antioxidant enzyme system; in addition, EGT can also inhibit the production of inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6, thereby protecting cells. Unlike other nutrients, EGT can efficiently enter cells and target mitochondria via its specific OCTN-1 transporter, rapidly clearing mitochondrial-specific ROS. Furthermore, EGT has been reported to have therapeutic effects on diseases related to mitochondrial damage, such as heart disease, Alzheimer's disease, and preeclampsia.

[0005] However, research on the effects of EGT on germ cells is scarce. Currently, only a few in vitro studies have been conducted on normal oocytes from livestock such as cattle and pigs. These studies all indicate that EGT can improve in vitro maturation and embryonic development of livestock oocytes by reducing oxidative stress. There are currently no studies or products related to the effects of EGT on older oocytes. Summary of the Invention

[0006] This invention provides the application of ergothionein in enhancing the fertility of older females, and develops ergothionein into a product that enhances the fertility of older females.

[0007] This invention provides the application of ergothionein in the preparation of products that enhance the fertility of older females.

[0008] In a preferred embodiment of the present invention, the type of product includes pharmaceuticals or food.

[0009] In a preferred embodiment of the present invention, the dosage form of the product includes an oral preparation.

[0010] In a preferred embodiment of the present invention, the improvement of fertility in older female individuals includes at least one of the following: (1) improving the in vitro fertilization rate of oocytes in older female individuals;

[0011] (2) Improve the blastocyst rate of oocytes from older female individuals;

[0012] (3) Increase the number of superovulations in older females;

[0013] (4) Increase the number of offspring produced by older females.

[0014] The present invention also provides a drug for enhancing the fertility of older females, the active ingredient of which includes ergothioneine, and also includes pharmaceutically acceptable excipients.

[0015] In a preferred embodiment of the present invention, the working concentration of ergothioneine in the drug, calculated in mice, is not less than 10 mg / kg.

[0016] The present invention also provides a food containing ergothioneine.

[0017] In a preferred embodiment of the present invention, the working concentration of ergothionein in the food, calculated in mice, is not less than 10 mg / kg.

[0018] Beneficial Effects: This invention provides the application of EGT in the preparation of products that enhance the fertility of older females. In the embodiments of this invention, older ICR female mice were used as animal models to verify the effects of EGT on the fertility of older female mice from both in vivo and in vitro perspectives. The results showed that EGT increased the in vitro fertilization rate and blastocyst rate of oocytes from older female mice; and increased the number of superovulations and litters in older female mice. This demonstrates that the EGT described in this invention has an improving effect on the fertility of older females. Attached Figure Description

[0019] Figure 1The figures show the in vitro fertilization and development of MII stage oocytes in young, aged, and aged + EGT groups after 10 days of EGT gavage treatment. Figure a: Schematic diagram of drug administration, in vitro fertilization and in vitro culture development of MII stage oocytes in young, aged, and aged + EGT groups; b: Representative figures of MII stage oocytes from young, aged, aged + 10 mg / kg EGT, and aged + 50 mg / kg EGT groups, showing in vitro fertilization and development to 2-cell, 4-8-cell, and blastocyst stages; c: In vitro fertilization rate of MII stage oocytes in young, aged, aged + 10 mg / kg EGT, and aged + 50 mg / kg EGT groups; d: Rate of MII stage oocytes from young, aged, aged + 10 mg / kg EGT, and aged + 50 mg / kg EGT groups reaching blastocyst stage in vitro.

[0020] Figure 2 The figures show the superovulation and litter size of mice in the Young, Aged, and Aged+EGT groups after 10 days of EGT gavage treatment. Figure a: Schematic diagram of EGT treatment, superovulation, mating, and litter size in the Young, Aged, and Aged+EGT groups; b: Representative litter size of female mice in the Young, Aged, Aged+10mg / kg EGT, and Aged+50mg / kg EGT groups; c: Litter size of female mice in the Young, Aged, Aged+10mg / kg EGT, and Aged+50mg / kg EGT groups; d: Number of superovulations in female mice in the Young, Aged, Aged+10mg / kg EGT, and Aged+50mg / kg EGT groups. Detailed Implementation

[0021] This invention provides the application of ergothionein in the preparation of products that enhance the fertility of older females.

[0022] The ergothioneine (EGT) described in this invention has the structure shown in Formula I, with the molecular formula: C9H 15 N3O2S, molecular weight: 229.3, CAS number: 497-30-3.

[0023]

[0024] This invention does not specifically limit the type of the product; the EGT can be prepared into a pharmaceutical or food product. This invention also does not specifically limit the dosage form of the product; it can be an oral preparation.

[0025] In this embodiment of the invention, high-fertility ICR female mice were used as animal models to verify the effect of EGT on the fertility of high-fertility female mice from both in vivo and in vitro perspectives. The results showed that EGT can increase the in vitro fertilization rate and blastocyst rate of oocytes in high-fertility female mice; and increase the number of superovulations and litters in high-fertility female mice.

[0026] The advanced reproductive age females described in this invention are required to be over 35 years old in humans and 8-10 months old in mice.

[0027] The present invention also provides a drug for enhancing the fertility of older females, the active ingredient of which includes ergothioneine, and also includes pharmaceutically acceptable excipients.

[0028] The present invention does not have any particular limitation on the dosage form of the drug, such as oral liquid. In the embodiments of the present invention, the effects of different concentrations of EGT on high-fertility ICR female mice were verified. It was found that significant differences could be observed at a dosage of 10 mg / kg. Therefore, the working concentration of ergothioneine in the drug, calculated in mice, is not less than 10 mg / kg.

[0029] The present invention also provides a food containing ergothioneine.

[0030] The working concentration of ergothioneine in the food product described in this invention, calculated in mice, is not less than 10 mg / kg. This invention does not specifically limit the type of food product; it only requires that EGT be added to an edible food product at the required dosage.

[0031] To further illustrate the present invention, the application of ergothionein provided by the present invention in enhancing the fertility of older females is described in detail below with reference to embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0032] Example 1

[0033] Using older, ICR female mice as an animal model, this study demonstrated, both in vivo and in vitro, that EGT can increase the in vitro fertilization rate and blastocyst rate of older oocytes; and increase the number of superovulations and litters in older female mice. The specific experimental procedure is as follows:

[0034] 1. Evaluation of in vitro fertilization and development of oocytes

[0035] Adult female ICR mice aged 8–10 weeks and 8–10 months were selected to represent young and advanced reproductive-age females, respectively. Advanced reproductive-age female mice were administered EGT at doses of 10 mg / kg or 50 mg / kg via gavage for 10 consecutive days, while the young and advanced reproductive-age control groups were administered the corresponding volume of physiological saline via gavage. After the administration period, PMSG and hCG were injected intraperitoneally sequentially to induce superovulation.

[0036] according to Figure 1 As shown in Figure a, MII stage oocytes from female mice in the Young, Aged, and Aged+EGT groups were collected, and in vitro fertilization and in vitro culture were performed. The in vitro fertilization rate and blastocyst rate of oocytes in each group were examined and statistically analyzed.

[0037] The test results showed that: ① Compared with the young group, the in vitro fertilization rate of the aged group was significantly lower, suggesting that advanced reproductive age reduces the fertilization rate of oocytes. However, the in vitro fertilization rate of the aged + EGT group treated with different doses of EGT by gavage was significantly increased, and there were significant differences between the low and high concentration EGT groups. Figure 1 (c). ② Compared with the younger group, the blastocyst rate of older mice was significantly decreased, suggesting that older age hinders the later development of fertilized eggs. However, the blastocyst rate was significantly increased in the older + EGT group treated with different doses of EGT by gavage, and there were significant differences between the low and high concentration EGT groups. Figure 1 (d) indicates that EGT can improve the later development process of fertilized eggs from older women.

[0038] 2. In vivo evaluation of the effect of EGT on the quality of advanced maternal age oocytes

[0039] This invention investigated the effects of EGT on the number of superovulations and litter size in older female mice, directly reflecting the effect of EGT on female fertility. Adult female ICR mice aged 8–10 weeks and 8–10 months were selected, corresponding to young and older females, respectively. Older female mice were administered EGT at doses of 10 mg / kg or 50 mg / kg via gavage for 10 consecutive days. After administration, some female mice were subsequently injected intraperitoneally with PMSG and hCG to induce superovulation. MII oocytes were collected from the young, older, and older+EGT groups to count the number of ovulations. Other female mice were mated 1:1 with age-appropriate males for one day, and then housed separately for approximately 20 days until litter size. Litter size was then recorded for each group. Figure 2 (a)

[0040] The experimental results showed that the litter size of older female mice was significantly smaller than that of younger mice. However, after treatment with different doses of EGT, the litter size of older female mice increased to varying degrees, and the differences were statistically significant. Figure 2 (b and c). For example Figure 2As shown in Figure d, the number of superovulated eggs in older female mice was significantly lower than that in younger mice. However, after treatment with different doses of EGT, the number of superovulated eggs in older female mice increased to varying degrees, and all showed significant differences compared with the control group. This suggests that EGT can enhance the fertility of older female mice.

[0041] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of ergothionein in the preparation of products that enhance the fertility of older females.

2. The application according to claim 1, characterized in that, The types of products mentioned include pharmaceuticals or food.

3. The application according to claim 1 or 2, characterized in that, The product can be in oral dosage form.

4. The application according to claim 1, characterized in that, The improvement of fertility in older female individuals includes at least one of the following: (1) improving the in vitro fertilization rate of oocytes in older female individuals; (2) Improve the blastocyst rate of oocytes from older female individuals; (3) Increase the number of superovulations in older females; (4) Increase the number of offspring produced by older females.

5. A drug for enhancing the fertility of older females, characterized in that, The active ingredient includes ergothioneine, and also includes pharmaceutically acceptable excipients.

6. The drug according to claim 5, characterized in that, The working concentration of ergothionein in the drug, calculated in mice, shall not be less than 10 mg / kg.

7. A food containing ergothioneine.

8. The food product according to claim 7, characterized in that, The working concentration of ergothionein in the food, calculated in mice, shall not be less than 10 mg / kg.

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