Application of nicotinamide ribose in preparation of medicine for increasing number of primordial germ cells and prolonging growth cycle

Abstract

The invention provides application of nicotinamide ribose in preparation of drugs for increasing the number of primordial germ cells and prolonging the growth cycle, and relates to the technical field of cell biology. Animal experiments prove that the supplement of nicotinamide ribose in the embryonic period can significantly increase the number of PGCs and prolong the post-birth growth cycle, and the action mechanism comprises: nicotinamide ribose promotes the proliferation of embryonic cells by up-regulating the NAD + level in PGCs, and enhances the expression of TFAP2C at the same time to maintain the undifferentiated state of cells and delay the meiosis initiation of female embryo PGCs and the differentiation initiation of male embryo PGCs; in post-birth effect evaluation, the number of follicles in ovaries of the female child rats in the NR group is remarkably increased, the fertility cycle is remarkably prolonged, the fertility of filial generations is not affected, and nicotinamide ribose is prompted to effectively maintain reproductive reserve and enhance the fertility by optimizing the development time sequence of PGCs. The invention provides a solid experimental foundation for the application of nicotinamide ribose in increasing the number of primordial germ cells and prolonging the growth cycle, and has a wide application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cell biology, and particularly relates to application of nicotinamide riboside in preparation of a medicine for increasing the number of primordial germ cells and prolonging the reproductive period. BACKGROUND

[0002] With the decline of global fertility rate and the delay of reproductive age, the decline of fertility caused by the decline of ovarian / testicular function has become a major social problem. According to the report of the United Nations World Population Prospects 2022, the global total fertility rate has dropped from 4.7 in 1950 to 2.4 in 2020, and more than half of the countries have a fertility rate below the population replacement level. At the same time, the average age of first-time childbirth of women in developed countries is generally delayed to over 30 years old (31.3 years old in Europe on average), and the sperm concentration of men has decreased by 1.4% per year in the past 40 years, forming a vicious cycle of reproductive aging and gamete quality degradation. The number and quality of primordial germ cells (PGCs) directly determine the individual's lifelong fertility. Abnormal proliferation and differentiation of PGCs during embryonic period will lead to premature depletion of postnatal reproductive reserve. Studies have shown that a decrease in the number of PGCs during embryonic period can cause premature depletion of postnatal ovarian reserve; human clinical data also reveals that about 30% of premature ovarian failure cases are related to developmental defects of PGCs.

[0003] At present, there are limited effective intervention means for the reduction or degradation of reproductive cells. Existing treatment methods mainly include hormone replacement, assisted reproductive technology, microsurgery, drug therapy and stem cell therapy, etc. However, these methods have limitations such as the risk of tumor in hormone replacement, high cost and ethical controversy in assisted reproductive technology, complexity in microsurgery, etc. Compared with these methods, drug therapy has higher safety, simpler operation and wider public acceptance, and thus becomes a more potential intervention strategy. For example, the patent with publication number CN104946582A invents a substance for promoting proliferation of chicken primordial germ cells, but its application is limited to in vitro culture system, and its in vivo efficacy is not clear, so its potential impact on the fertility of postnatal individuals cannot be evaluated.

[0004] Nicotinamide adenine dinucleotide (NAD + ) is a key cofactor indispensable for cellular energy metabolism and signal transduction, and its homeostasis is maintained through de novo synthesis and salvage synthesis pathways. In recent years, the NAD + metabolic pathway has attracted attention due to its central role in anti-aging and cellular energy regulation. As an important precursor of NAD + , nicotinamide riboside (NR) has the effects of improving mitochondrial function and delaying aging, and has high safety and good absorption and metabolism characteristics in vivo. Studies have shown that NR has intervention potential in various aging-related disease models: in neurodegenerative diseases, NR improves NAD +Nicotinamide ribose (NR) can slow the progression of Parkinson's disease; in cardiovascular diseases, it can significantly improve exercise capacity in patients with peripheral artery disease; in metabolic diseases, NR can alleviate diabetes and non-alcoholic fatty liver disease by regulating insulin sensitivity and improving liver function. Although the role of NR in cancer treatment is not yet clear, its ability to improve energy metabolism and enhance DNA repair may have potential benefits for certain cancer types. However, the regulatory mechanisms and roles of NR in PGC development have not been reported. Based on this, this invention proposes the application of nicotinamide ribose in the preparation of drugs that increase the number of primordial germ cells and prolong the reproductive cycle, aiming to provide a novel intervention strategy for reproductive health and a new approach to developing drugs that increase the number of PGCs and prolong the reproductive cycle. Summary of the Invention

[0005] (a) Technical problems to be solved

[0006] To address the shortcomings of existing technologies, this invention provides the application of nicotinamide ribose in the preparation of drugs that increase the number of primordial germ cells and prolong the reproductive cycle.

[0007] (II) Technical Solution

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] This invention provides the use of nicotinamide ribose in the preparation of pharmaceutical or health products for increasing the number of primordial germ cells, the use including increasing the number of primordial germ cells in the genital ridge during embryonic development and increasing the number of female and male germ cells after birth. The structural formula of the nicotinamide ribose is as follows: Figure 1 As shown.

[0010] This invention provides the use of nicotinamide ribose in the preparation of a drug for promoting the proliferation of primordial germ cells.

[0011] This invention provides the use of nicotinamide ribose in the preparation of a medicament for maintaining the undifferentiated state of primordial germ cells.

[0012] This invention provides the use of nicotinamide ribose in the preparation of a medicament for delaying the meiotic process of female embryonic primordial germ cells, the process including the initiation and subsequent developmental stages of meiosis.

[0013] This invention provides the use of nicotinamide ribose in the preparation of a medicament for delaying the initiation of differentiation of male embryonic primordial germ cells.

[0014] This invention provides the application of nicotinamide ribose in the preparation of drugs for regulating mitochondrial function and energy metabolism in primordial germ cells.

[0015] This invention provides the use of nicotinamide ribose in the preparation of a drug for increasing the follicular reserve in the ovaries of offspring, wherein the follicles include total follicles, primordial follicles, primary follicles, secondary follicles, antral follicles, and atretic follicles.

[0016] This invention provides the use of nicotinamide ribose in the preparation of a medicament for prolonging the reproductive cycle, wherein the prolongation is achieved by improving the fertility of offspring.

[0017] (III) Beneficial Effects

[0018] This invention provides the application of nicotinamide ribose in the preparation of drugs that increase the number of primordial germ cells and prolong the reproductive cycle. Animal experiments have confirmed that nicotinamide ribose supplementation during the embryonic period can significantly increase the number of primordial germ cells and prolong the postnatal reproductive cycle. Its mechanism of action includes: nicotinamide ribose upregulates NAD+ in primordial germ cells. + Nicotinamide ribose promotes the proliferation of primordial germ cells during the embryonic period, while enhancing TFAP2C expression to maintain the undifferentiated state of cells, delaying the initiation of meiosis in female embryonic primordial germ cells and the initiation of differentiation in male embryonic primordial germ cells. In postnatal effect assessment, the number of follicles in the ovaries of female offspring in the NR group was significantly increased, the reproductive cycle was significantly prolonged, and the fertility of offspring was not affected. This indicates that nicotinamide ribose can effectively maintain reproductive reserve and enhance fertility by optimizing the developmental timing of PGCs. This invention provides a solid experimental basis for the application of nicotinamide ribose in increasing the number of primordial germ cells and prolonging the reproductive cycle, and has broad application prospects. Attached Figure Description

[0019] Figure 1 This is the chemical structural formula of nicotinamide ribose.

[0020] Figure 2 Results of nicotinamide riboside supplementation during pregnancy increasing the number of progenitor cells (PGCs) in the embryonic stage. Figure 2 A represents the NAD of PGCs in E13.5 embryos. + Horizontal statistical results; Figure 2 B is a statistical chart showing the number of PGCs sorted by FACS for E13.5 and E15.5 embryos; Figure 2 C is an immunofluorescence image of the entire reproductive ridge of an E13.5 embryo; Figure 2 D is a statistical graph of immunofluorescence staining of the whole organs of the gonadal ridge in female embryos of E12.5, E13.5 and E15.5. Figure 2 E represents the statistical graph of immunofluorescence staining of the whole organs of the reproductive ridge in male embryos of E12.5, E13.5, and E15.5.

[0021] Figure 3 The results of nicotinamide riboside supplementation during pregnancy increasing the number of germ cells after birth. Figure 3A is a VASA immunofluorescence image of a section of the ovary of a female P5 mouse. Figure 3 B is a statistical graph showing the number of germ cells in the ovaries of female P5 mice; Figure 3 C is a statistical graph showing the average number of germ cells per seminiferous tubule in the testes of male P5 mice.

[0022] Figure 4 The results of nicotinamide riboside supplementation during pregnancy promoting the proliferation of E13.5 PGCs. Figure 4 A is an immunofluorescence image of VASA and BrdU on a section of the genital ridge of an E13.5 embryo; Figure 4 B is a statistical graph showing the BrdU positivity rate in PGCs from female E13.5 embryos; Figure 4 C is a statistical graph showing the BrdU positivity rate in PGCs from male embryos of E13.5.

[0023] Figure 4 D represents the RT-qPCR results of key cell cycle-related genes in E13.5 female embryo PGCs; Figure 4 E represents the RT-qPCR results of key cell cycle-related genes in PGCs from male embryos of E13.5.

[0024] Figure 5 Results of nicotinamide riboside supplementation during pregnancy to maintain the undifferentiated state of E13.5 PGCs. Figure 5 A is a VASA and TFAP2C immunofluorescence image of a section of the genital ridge of an E13.5 embryo; Figure 5 B is a statistical graph of TFAP2C fluorescence intensity in PGCs of female E13.5 embryos; Figure 5 C is a statistical graph of TFAP2C fluorescence intensity in PGCs of male embryos from E13.5.

[0025] Figure 6 The results of nicotinamide riboside supplementation during pregnancy to delay the initiation of meiosis in PGCs of E15.5 female embryos. Figure 6 A is an VASA and SYCP3 immunofluorescence image of a section of the genital ridge of an E15.5 female embryo; Figure 6 B is an immunofluorescence image of VASA and γH2AX from a section of the genital ridge of an E15.5 female embryo; Figure 6 C is a statistical graph showing the SYCP3 positivity rate in PGCs of E15.5 female embryos; Figure 6 D is a statistical graph showing the positivity rate of γH2AX in PGCs of female E15.5 embryos; Figure 6 E represents the RT-qPCR results of key meiosis-related genes in PGCs of female embryos of E13.5.

[0026] Figure 7The results of nicotinamide riboside supplementation during pregnancy delayed the meiotic process of PGCs in female embryos from E16.5 to E18.5. Figure 7 A shows immunofluorescence images of SYCP3 and γH2AX spreading on the genital ridge of female embryos at E16.5, E17.5, and E18.5 (Leptotene: Leptotene stage, Zygotene: Zygotene stage, Pachytene: Pachytene stage, Diplotene: Diplotene stage). Figure 7 B is a statistical graph showing the proportion of each stage of prophase I in female embryo PGCs from E16.5, E17.5, and E18.5 embryos.

[0027] Figure 8 The results of nicotinamide riboside supplementation during pregnancy delayed the differentiation of progenitor cells (PGCs) in E15.5 male embryos. Figure 8 A shows VASA and MILI immunofluorescence images of a section of the genital ridge of an E15.5 male embryo; Figure 8 B is a statistical graph showing the MILI positivity rate in PGCs from male E15.5 embryos; Figure 8 C represents the RT-qPCR results of key differentiation-related genes in PGCs from male E13.5 embryos.

[0028] Figure 9 The long-term effects of nicotinamide riboside supplementation during pregnancy on improving ovarian reserve in offspring. Figure 9 Image A shows HE staining of the ovaries of female mice at 3M, 8M, 12M, and 16M. Figure 9 B is a statistical chart showing the number of follicles in the ovary at 3M, 8M, 12M, and 16M.

[0029] Figure 10 The results of supplementing pregnant women with nicotinamide riboside to improve offspring fertility. Figure 10 A is a statistical graph showing the weight of female mouse ovaries; Figure 10 B is a statistical graph showing the changes in AMH levels of female offspring over time; Figure 10 C is a statistical graph showing the cumulative number of offspring born to female mice over time; Figure 10 D is a statistical graph showing the total number of offspring per female mouse during the entire testing period; Figure 10 E is a statistical chart showing the total number of offspring born to female mice; Figure 10 F shows a representative image of each litter of female mice. Detailed Implementation

[0030] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] Example 1

[0032] Eighty two-month-old female C57BL / 6j pregnant mice were randomly divided into two groups: a control group (WT group) and a nicotinamide-ribose supplemented group (NR group), with 40 mice in each group. The WT group was fed a basal diet, while the NR group was fed a basal diet supplemented with 400 mg / kg / day of nicotinamide-ribose. Both groups were fed this diet continuously until the end of pregnancy.

[0033] Three pregnant mice were randomly selected from each group. On day 13.5 (E13.5), after deep anesthesia with isoflurane, the uterine horn was dissected under aseptic conditions, the germinal crest tissue of the embryo was separated, primordial germ cells (PGCs) were sorted by flow cytometry (FACS), and their NAD values ​​were measured. + Content: The number of PGCs in the reproductive ridge was counted by whole-organ immunofluorescence combined with VASA staining for germ cell markers at E12.5, E13.5, and E15.5, and quantified by FACS sorting.

[0034] Experimental results are as follows Figure 2 As shown, by Figure 2 As shown in Figure A, compared with the WT group, the NR group mice had higher levels of NAD+ in E13.5 PGCs. + The content increased significantly (P<0.05); by Figure 2 As shown in B, the number of PGCs obtained by the NR group through FACS sorting at E13.5 was not significantly different from that of the WT group (P>0.05), but the number of PGCs was significantly greater in the NR group at E15.5 than in the WT group (P<0.05). Figure 2 According to CE, the results of whole-organ immunofluorescence staining showed that at E15.5, the number of PGCs in the genital ridge of the NR group was significantly higher than that of the WT group.

[0035] The results in summary indicate that nicotinamide riboside supplementation during the embryonic period can upregulate intracellular NAD in PGCs. + The level promotes its proliferation at E13.5 and significantly increases the number of PGCs at E15.5.

[0036] Example 2

[0037] Three pregnant mice were randomly selected from each group in Example 1. Immediately after natural delivery, the NR group diet was changed to the basal diet and continued until the sampling. On day 5 after birth (P5), three female and three male pups were randomly selected from each group for sampling. Ovaries of female mice and testes of male mice were collected. The tissue samples were fixed in 4% formaldehyde, embedded in paraffin, and serially sectioned (5 μm thick). The sections were stained with VASA, a germ cell marker, and the number of germ cells was observed and counted under a fluorescence microscope.

[0038] Experimental results are as follows Figure 3 As shown, from Figure 3 A and B indicate that the number of germ cells in the ovaries of female F1 offspring in the NR group was significantly higher than that in the WT group (P<0.05); Figure 3 As shown in A and C, the average number of germ cells per seminiferous tubule in the testes of male F1 generation mice in the NR group was significantly higher than that in the WT group (P<0.05).

[0039] In summary, the experimental results indicate that nicotinamide riboside supplementation during the embryonic period can increase the number of germ cells in both female and male mice after birth.

[0040] Example 3

[0041] Three pregnant mice were randomly selected from each group in Example 1 and injected intraperitoneally with 5-bromo-2'-deoxyuridine (BrdU, 150 mg / kg) at E13.5. Three hours later, the mice were euthanized by cervical dislocation, and the uterine horns were rapidly dissected to separate male and female embryos. Three female embryos and three male embryos were randomly selected from each group. The ovaries of the female embryos and the testes of the male embryos were separated. Tissue samples were fixed in 4% formaldehyde, embedded in paraffin, and serially sectioned (5 μm thick). After co-staining with BrdU and VASA immunofluorescence, the sections were observed under an immunofluorescence microscope, and the positive rate of BrdU in PGCs was calculated. The expression levels of key cell cycle-related genes Cdk1, Cdk2ap2, Cks2, Eif4ebp1, Ube2c, and Ube2s in embryonic PGCs were detected by RT-qPCR.

[0042] Experimental results are as follows Figure 4 As shown, from Figure 4 AC analysis revealed that, compared to the WT group, the positive rate of BrdU in PGCs of female embryos in the NR group was significantly increased (P<0.05), and the positive rate of BrdU in PGCs of male embryos was significantly increased (P<0.05). Further analysis using RT-qPCR revealed the increased expression of cell cycle regulatory genes. Figure 4 DE) revealed that the expression levels of key cell cycle-related genes Cdk1, Cdk2ap2, Cks2, Eif4ebp1, Ube2c, and Ube2s in NR group embryo PGCs were significantly upregulated compared to WT group (P<0.05).

[0043] In summary, the experimental results indicate that nicotinamide ribose supplementation during the embryonic period can promote the proliferation of primordial germ cells in both female and male embryos.

[0044] Example 4

[0045] Three pregnant mice were randomly selected from each group in Example 1. After deep anesthesia with isoflurane at E13.5, the uterine horns were dissected under aseptic conditions, and male and female embryos were separated. Three female embryos and three male embryos were randomly selected from each group, and the genital ridge tissue was separated, embedded in paraffin, and serially sectioned (5 μm thick). The sections were co-stained with TFAP2C and VASA immunofluorescence, and the fluorescence intensity of TFAP2C in PGCs was observed and counted under an immunofluorescence microscope.

[0046] Experimental results are as follows Figure 5 As shown, compared with the WT group, the fluorescence intensity of TFAP2C in PGCs of both female and male embryos in the NR group was significantly increased (P<0.05), indicating that nicotinamide riboside supplementation during the embryonic period can maintain the undifferentiated state of PGCs by upregulating TFAP2C expression.

[0047] Example 5

[0048] Three pregnant mice were randomly selected from each group in Example 1. After deep anesthesia with isoflurane at E15.5, the uterine horns were dissected under aseptic conditions, and male and female embryos were separated. Three female embryos were randomly selected from each group, and the genital ridge tissue was separated, embedded in paraffin, and serially sectioned (5 μm thick). The sections were co-stained with immunofluorescence for SYCP3 and VASA and γH2AX and VASA, respectively. The positive rates of SYCP3 and γH2AX in PGCs were observed and counted under an immunofluorescence microscope. The expression levels of key meiosis-related genes Stra8, Rad51b, Dmc1, and Sycp1 in female embryonic PGCs were detected by RT-qPCR.

[0049] Experimental results are as follows Figure 6 As shown, from Figure 6 AD analysis showed that, compared with the WT group, the positive rates of SYCP3 and γH2AX in PGCs of female embryos in the NR group were significantly lower (P<0.05). Figure 6 E showed that the expression levels of key meiosis-related genes Stra8, Rad51b, Dmc1, and Sycp1 in female embryos of the NR group were significantly lower than those in the WT group (P<0.05).

[0050] In summary, the experimental results indicate that nicotinamide riboside supplementation during the embryonic period can significantly delay the process of female embryonic PGCs entering meiosis by inhibiting γH2AX and SYCP3, downregulating the expression of homologous recombination and synapsis-related genes.

[0051] Example 6

[0052] Nine pregnant mice were randomly selected from each group in Example 1. At E16.5, E17.5, and E18.5 (n=3 mice at each time point), the uterine horns were dissected under aseptic conditions, and male and female embryos were separated. Three female embryos were randomly selected from each group, and the genital ridge tissue was isolated. Primitive germ cells from the genital ridge were fixed on clean glass slides with 1% paraformaldehyde solution. The fixed cells were blocked using antibody dilution buffer (ADB), and immunofluorescence staining with SYCP3 and γH2AX was performed. Based on the staining morphology characteristics of SYCP3 and γH2AX, PGCs at each stage of prophase I of meiosis (including leptotene, zygotene, pachytene, and diplotene) were identified and counted. The proportion of PGCs at each time point in each stage and group was statistically analyzed.

[0053] Experimental results are as follows Figure 7 As shown, compared with the WT group, the NR group had a higher proportion of PGCs in the early stages of embryonic development (leptomeninges and pachytene stages) and a lower proportion of PGCs in the late stages (pachytene stages and diplotene stages). This indicates that nicotinamide riboside supplementation during the embryonic period significantly delays the temporal progression of female embryonic PGCs from the leptomeninges to the pachytene / diplotene stages by inhibiting SYCP3 and γH2AX in promeiosis, thereby delaying the meiotic process of female embryonic primordial germ cells.

[0054] Example 7

[0055] Three pregnant mice were randomly selected from each group in Example 1. After deep anesthesia with isoflurane at E15.5, the uterine horns were dissected under aseptic conditions, and male and female embryos were separated. Three male embryos were randomly selected from each group, and the genital ridge tissue was separated. The tissue samples were embedded in paraffin and then serially sectioned (5 μm thick). The sections were co-stained with MILI and VASA immunofluorescence. The positive rate of MILI in PGCs was observed and counted under an immunofluorescence microscope. The expression levels of key differentiation-related genes Piwil2, Tdrd5, Tex14, and Bnc2 in male embryo PGCs were detected by RT-qPCR.

[0056] Experimental results are as follows Figure 8 As shown, from Figure 8 A and B indicate that, compared with the WT group, the positive rate of MILI in PGCs of male embryos in the NR group was significantly lower (P<0.05); from Figure 8C indicates that the expression levels of key differentiation-related genes Piwil2, Tdrd5, Tex14, and Bnc2 in male embryo PGCs from the NR group were significantly downregulated compared to those in the WT group (P<0.05).

[0057] In summary, the experimental results indicate that nicotinamide ribose supplementation during the embryonic period significantly delays the initiation of differentiation of PGCs in male embryos by inhibiting MILI protein expression and core genes of the Piwi pathway.

[0058] Example 8

[0059] Three pregnant mice were randomly selected from each group in Example 1. Immediately after natural delivery, the NR group diet was replaced with the basal diet and continued until the tissue samples were collected. At 3, 8, 12, and 16 months after birth, three female mice from each group were randomly selected to isolate the ovaries. The tissue samples were embedded in paraffin and then serially sectioned (5 μm thick). The samples were stained with hematoxylin and eosin (HE) and the number of follicles at each stage (including primordial follicles (PMF), primary follicles (PF), secondary follicles (SF), antral follicles (AF), atretic follicles (Atretic), and total follicles (Total)) was counted to assess ovarian reserve capacity.

[0060] Experimental results are as follows Figure 9 As shown, during aging, the number of follicles in the ovaries of female F1 offspring mice in the WT group gradually decreased. In contrast, the number of follicles in female F1 offspring mice in the NR group was significantly higher than that in the WT group at all time points (P<0.05), indicating that nicotinamide ribose supplementation during the embryonic period can maintain and improve the ovarian reserve of offspring mice.

[0061] Example 9

[0062] Three pregnant mice were randomly selected from each group in Example 1. Immediately after natural delivery, the NR group diet was changed to the basal diet and continued until the harvest. From 2 months of age (2M), all offspring were subjected to a continuous co-breeding experiment, and the birthing status of each female mouse was recorded, including the average number of offspring per litter, the cumulative number of offspring, and the total number of offspring. Serum AMH levels were measured by ELISA at three time points: 3M, 8M, and 12M.

[0063] Experimental results are as follows Figure 10 As shown, from Figure 10 A indicates that while the ovarian weight of female mice in the NR group at 3M, 8M, and 12M was higher than that in the WT group, the differences were not statistically significant (P>0.05); however, at 16M, the ovarian weight of the NR group was significantly higher than that of the WT group (P<0.05). Figure 10 B indicates that, with increasing age, serum anti-Müllerian hormone (AMH) levels in F1 generation female mice of both the WT and NR groups showed a decreasing trend, but the AMH level in the NR group was significantly higher than that in the WT group at all time points; from Figure 10CE showed that the cumulative number of offspring and the total number of offspring in the NR group were significantly higher than those in the WT group (P<0.05).

[0064] In summary, the experimental results indicate that nicotinamide riboside supplementation during the embryonic period can effectively improve the fertility of older female mice by increasing the number of offspring germ cells and maintaining ovarian reserve function.

[0065] This invention utilizes a mouse model of embryonic development to demonstrate that nicotinamide riboside supplementation during embryonic development can significantly increase the number of primordial germ cells and prolong the postnatal reproductive cycle. Its mechanism of action includes: nicotinamide riboside upregulates NAD+ in primordial germ cells. + Nicotinamide ribose promotes the proliferation of primordial germ cells during the embryonic period, while enhancing TFAP2C expression to maintain the undifferentiated state of cells, delaying the initiation of meiosis in female embryonic primordial germ cells and the initiation of differentiation in male embryonic primordial germ cells. In postnatal effect assessment, the number of primordial follicles, primary follicles, secondary follicles, antral follicles, and total follicles in the ovaries of female offspring in the NR group were significantly increased, the reproductive cycle was significantly prolonged, and the fertility of offspring did not decline, suggesting that nicotinamide ribose effectively maintains reproductive reserve and enhances fertility by optimizing the developmental sequence of PGCs. This invention provides a solid experimental basis for the application of nicotinamide ribose in increasing the number of primordial germ cells and prolonging the reproductive cycle, and has broad application prospects.

[0066] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The use of nicotinamide ribose in the preparation of pharmaceuticals or health supplements for increasing the number of primordial germ cells, characterized in that, The applications include increasing the number of primordial germ cells in the genital ridge during embryonic development and increasing the number of female and male germ cells after birth.

2. Application of nicotinamide ribose in the preparation of drugs for promoting the proliferation of primordial germ cells.

3. Application of nicotinamide ribose in the preparation of drugs for maintaining the undifferentiated state of primordial germ cells.

4. The use of nicotinamide ribose in the preparation of drugs for delaying the meiotic process of female embryonic primordial germ cells, characterized in that, The process includes the initiation of meiosis and subsequent developmental stages.

5. Application of nicotinamide ribose in the preparation of drugs for delaying the initiation of differentiation of male embryonic primordial germ cells.

6. Application of nicotinamide ribose in the preparation of drugs for regulating mitochondrial function and energy metabolism in primordial germ cells.

7. The application of nicotinamide ribose in the preparation of drugs for increasing the follicular reserve in the ovaries of offspring, characterized in that, The follicles include total follicles, primordial follicles, primary follicles, secondary follicles, antral follicles, and atretic follicles.

8. Application of nicotinamide ribose in the preparation of drugs for prolonging the reproductive cycle.

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