Application of agar polysaccharide degradation products in the preparation of drugs for improving male fertility

Abstract

This invention discloses the use of agar polysaccharide degradation products in the preparation of a drug for improving male fertility. The method of the invention yields a compound that can be used to enhance sperm motility and male fertility, providing new insights and clues for further research into the pathogenesis of oligoasthenozoospermia and male reproductive health.

Description

Technical field:

[0001] The invention relates to the application of agar polysaccharide degradation products in the preparation of drugs for improving male fertility, and belongs to the fields of reproductive biology technology and biomedicine. Background technology:

[0002] According to the latest report from the World Health Organization, approximately 17.5% (about one-sixth) of the adult population worldwide is affected by infertility. Several studies have shown that there are many factors contributing to infertility, of which male factors account for approximately 50%. With changes in the social environment and increased social pressure, male fertility has declined, and the incidence of infertility has gradually increased. Among them, decreased sperm quality is one of the key issues of concern in the field of reproductive medicine today. A 2022 study from the Hebrew University of Israel showed that the average sperm count and concentration of men worldwide has decreased by 62% over the past 45 years. Furthermore, the team also pointed out that the decline in the average sperm count of men worldwide will continue to accelerate in the 21st century. Faced with such a serious situation, researchers must take action as soon as possible to protect male fertility and prevent further damage to male reproductive health.

[0003] Spermatogenesis is a complex and highly coordinated process in the male reproductive system, occurring in the seminiferous tubules of the testicles throughout the reproductive cycle. It involves spermatogonia differentiation, the initiation of meiosis, the initiation of spermatid elongation, and the release of sperm into the vas deferens. Spermatogenesis is a key component of male fertility, and its normal progression is essential for male fertility. Any abnormality in spermatogenesis can lead to male infertility. Therefore, maintaining sperm health is crucial for protecting male fertility.

[0004] Red algae, a type of algae primarily found in the ocean, are highly nutritious and offer numerous health benefits, playing a crucial role in ecosystems. Agar-oligosaccharides, a polysaccharide extracted from the cell walls of red algae, are primarily composed of agarose and agaropectin. Due to their unique gelling properties, they have been widely used in the food and biopharmaceutical industries. Agar-oligosaccharides, a hydrolyzed product of agar-oligosaccharides, exhibit greater water solubility and bioactivity than agar-oligosaccharides. They have shown great potential in anti-cancer, anti-inflammatory, and immune-enhancing activities. Furthermore, agar-oligosaccharides, as a novel prebiotic, can selectively promote the growth of two beneficial bacteria, bifidobacteria and lactobacilli, in the host's intestines, thereby promoting host health. This property contributes to overall health and can also positively support male reproductive health. Furthermore, multiple studies have demonstrated that agar-oligosaccharides possess antioxidant activity, scavenging free radicals and increasing the activity of antioxidant enzymes. Oxidative stress is closely linked to decreased male fertility, so the antioxidant activity of agar-oligosaccharides can help maintain sperm health. Summary of the invention:

[0005] The purpose of the present invention is to provide an effective treatment method for poor male sperm quality and low fertility, namely, the use of agar polysaccharide degradation products in the preparation of drugs for improving male fertility.

[0006] In the present invention, a male mouse model of oligoasthenozoospermia was established by intraperitoneal injection of busulfan, and it was found that the addition of agar oligosaccharide, a degradation product of agar polysaccharide, could significantly improve the sperm motility and semen density of male mice.

[0007] Therefore, the present invention provides the use of agar polysaccharide degradation products in the preparation of drugs for improving male fertility.

[0008] Preferably, the agar polysaccharide degradation product is agar oligosaccharide.

[0009] Preferably, the drug for improving male fertility is a drug for improving sperm motility and semen density.

[0010] Preferably, the drug contains a therapeutically effective amount of agar polysaccharide degradation product and an acceptable carrier.

[0011] The present invention also provides a medicine for improving male fertility, which contains an effective amount of agar polysaccharide degradation product.

[0012] The method of the present invention obtains a compound that can be used to improve sperm motility and male fertility, providing new ideas and clues for future in-depth research on the pathogenesis of oligoasthenozoospermia and for researchers of male reproductive health. Description of the drawings:

[0013] Figure 1 A conceptual diagram of the research conducted for this invention.

[0014] Figure 2 The results of sperm motility test of three groups of mouse samples are shown in Figure 2.

[0015] Figure 3 Statistical results of semen density of three groups of mouse samples. Specific implementation method:

[0016] The method of the present invention will be further described below with reference to the accompanying drawings and through specific embodiments.

[0017] Example 1

[0018] 1. Experimental animal treatment and reagent preparation

[0019] The present invention uses 3-week-old ICR male mice as experimental subjects. The mice are caged in a clean animal room and raised strictly according to standard mouse breeding conditions. The mice are placed in a 12-hour light and 12-hour dark cycle, the room temperature is 23±1°C, the ambient humidity is 50%, and they can drink water and eat freely. All procedures comply with animal ethics standards and experimental specifications. A normal control group (Con), a busulfan-treated group (Bus), and an agar oligosaccharide (purchased from Qingdao Bozhi Huili Biotechnology Co., Ltd., product batch number: 2023010401QBF) rescue group (Bus+Ao) were set up. Specific treatment method for mice: This experiment used 3-week-old ICR male mice. The Con group received a single intraperitoneal injection of normal saline at a dose of 7.5 μL per gram of body weight on the morning of the first day, and then continued to receive a single oral gavage of normal saline at a dose of 7.5 μL per gram of body weight every morning on the second day for five weeks; the Bus group received a single intraperitoneal injection of 1 mg / mL busulfan according to body weight on the morning of the first day, and then continued to receive a single oral gavage of normal saline at a dose of 7.5 μL per gram of body weight every morning on the second day for five weeks; the Bus+Ao group received a single intraperitoneal injection of 1 mg / mL busulfan according to body weight on the morning of the first day, and then continued to receive a single oral gavage of 40 mg / mL agar oligosaccharide according to body weight every morning on the second day for five weeks.

[0020] Preparation of busulfan solution: Dissolve 0.1 g of busulfan (B2635, Sigma, USA) in 10 mL of DMSO. Aliquot the 10 mg / mL busulfan solution into 1.5 mL Eppendorf tubes and store in the dark at 4°C until use. Dilute 10-fold with sterile saline before use (final concentration: 1 mg / mL). Intraperitoneally inject the solution using a 1 mL syringe, ensuring that the mouse receives 30 mg of busulfan per kg of body weight.

[0021] Agar oligosaccharide solution preparation: Dissolve agar oligosaccharide powder in sterile saline to a concentration of 40 mg / mL. Administer intragastrically daily in the morning for 5 weeks. Use a 1 mL syringe to ensure mice receive 300 mg of agar oligosaccharide per kg of body weight per day.

[0022] 2. Sample collection

[0023] After five weeks of continuous gavage treatment, testicular and semen samples were collected from the different treatment groups for comparative testing. Mice were sacrificed by cervical dislocation. The abdominal skin was sprayed with alcohol, and the epididymis was exposed with sterile surgical scissors. The epididymis was removed, and sperm was released for in vitro capacitation and sperm motility measurements.

[0024] 3. Sperm motility and semen density testing

[0025] Prepare sperm capacitation solution in advance by mixing α-MEM / F12 and FBS in a 9:1 volume ratio and preheating on a 37°C hot plate. Mice were sacrificed by cervical dislocation, and the cauda epididymis tissue was carefully removed and placed in a 3 cm dish containing 200 μL of sperm capacitation solution. The cauda epididymis was incised with a razor blade to release fresh sperm, and the mixture was timed for 5 minutes. Subsequently, 20 μL of the turbid solution was aspirated and diluted with 200 μL of fresh sperm capacitation solution, mixing thoroughly to obtain sperm fluid. Finally, 4 μL of the sperm fluid was aspirated and dropped onto a sperm slide to allow the sperm to quickly diffuse onto the slide. The sperm were then imaged using an SCA sperm quality analyzer for sperm motility and semen density analysis.

[0026] 4. Results

[0027] Figure 1 This diagram illustrates the research concept for this invention. Three-week-old male mice were given a single intraperitoneal injection of busulfan (1 mg / mL) to establish an oligoasthenozoospermia mouse model. Starting on the second day after successful model establishment, agar oligosaccharide (40 mg / mL) was administered orally daily for five weeks. Finally, we evaluated whether supplementing with agar oligosaccharide by gavage for five weeks after the busulfan injection could improve sperm motility and semen density. Figure 2 The results of sperm motility tests in three groups of mice show that compared with the control group, injection of busulfan (1 mg / mL) significantly decreased sperm motility (P<0.05). However, after five weeks of oral administration of agar oligosaccharide (40 mg / mL), sperm motility in mice was significantly improved compared with the busulfan group (P<0.05). Figure 3 The results show that compared with the control group, busulfan injection caused a significant decrease in semen density (P<0.05). However, after five weeks of oral administration of agar oligosaccharide, the semen density of mice was significantly increased compared with the busulfan group (P<0.05).

[0028] The method of the present invention obtains a compound that can be used to improve sperm motility and male fertility, providing new ideas and clues for future in-depth research on the pathogenesis of oligoasthenozoospermia and for researchers of male reproductive health.

Claims

1. The application of agar oligosaccharide in the preparation of a drug for improving male fertility, wherein the drug for improving male fertility is a drug for improving sperm motility and semen density.

2. The use according to claim 1, characterized in that The medicine contains an effective amount of agar oligosaccharide and an acceptable carrier.

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