Small peptide Phe-Gly-Phe capable of improving sperm motility level and mucus penetrating ability and application of small peptide Phe-Gly-Phe

Abstract

The invention relates to the technical field of biological medicines, in particular to a small peptide Phe-Gly-Phe capable of improving the sperm motility level and mucus penetrating capacity and application of the small peptide Phe-Gly-Phe, and it is found that the small peptide can improve motility and functions of human mature sperms under the in-vitro culture condition. In-vitro experiments prove that compared with a control group, after the Phe-Gly-Phe is added, the sperm motility level is obviously improved, and the mucus penetrating capacity is obviously improved. The application of Phe-Gly-Phe can effectively improve the functions of sperms in the in-vitro fertilization process by enhancing sperm motility and improving athletic ability, so that the fertilization success rate of sperms optimized or treated by the invention can be increased by means of artificial fertilization, in-vitro fertilization-embryo transplantation or single sperm injection in oocytoplasm and the like, infertility is treated, and the aim of preventing and treating infertility is fulfilled. The fertility outcome of the patient is improved to a great extent.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to a small peptide Phe-Gly-Phe for improving sperm motility and mucus penetration ability and an application thereof. Background Art

[0002] Sperm function, that is, the ability of sperm to fertilize under physiological conditions, is not only the key to ensuring normal fertilization, but also an important factor in measuring sperm quality. Sperm motility and mucus penetration are important indicators that reflect sperm function and evaluate male fertility.

[0003] The level of sperm motility is usually evaluated by sperm motility, which is an important factor affecting male fertilization ability. Sperm motility refers to the ability of sperm to move, and is usually evaluated by the proportion of active sperm to total sperm. The ability to move forward is crucial for sperm to achieve fertilization function, and refers to the proportion of sperm that move forward actively, linearly, and over a large range. High sperm motility means that sperm will have a higher chance of successfully passing through the female reproductive tract, meeting and combining with the egg, thereby increasing the success rate of conception. Therefore, if sperm wants to have normal survival and fertilization ability, it must have the ability to move forward.

[0004] Cervical mucus is an important substance in the female reproductive tract. It has a screening and filtering effect on sperm. Only sperm with strong vitality and normal morphology can successfully penetrate cervical mucus. The ability of sperm to penetrate mucus is a key step for sperm to reach the fallopian tube and combine with the egg.

[0005] Sperm quality is directly related to whether the sperm can successfully penetrate the cervical mucus, reach the fallopian tube and combine with the egg. It is of vital importance for conception. Sperm dysfunction is an important cause that affects sperm quality and leads to male infertility.

[0006] In summary, sperm motility directly reflects the motility and fertilization potential of sperm, which is of great significance for evaluating male fertility and is one of the main indicators for assessing semen quality. Low sperm motility can lead to asthenospermia, which is one of the most common causes of male infertility clinically, and currently about 50% of infertility is due to problems with the male spouse. Because of low sperm motility, sperm may not be able to reach the egg effectively, thus affecting conception. Even if it can combine with the egg and successfully fertilize, the poor quality of the fertilized egg will increase the probability of miscarriage and pregnancy complications during pregnancy, and increase the risk of fetal malformations or developmental delays.

[0007] For male infertile patients, infertility can be treated by improving sperm motility and mucus penetration through drug therapy and assisted reproductive technology. Currently, it is necessary to find certain drugs or physiologically active substances to improve sperm motility and sperm function to treat male infertility. For example, finding an active substance or treatment method that improves the ability to penetrate mucus is expected to become a new drug for the treatment of male infertility or a key step in in vitro assisted reproductive technology. Summary of the invention

[0008] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a small peptide Phe-Gly-Phe for improving sperm motility and mucus penetration ability and its application, which has significant effects, is reliable and safe, can effectively improve sperm motility, increase the ability to penetrate mucus, and is expected to treat male oligospermia and asthenospermia and improve sperm quality.

[0009] In order to achieve the above object, the present invention adopts the following technical solutions:

[0010] A small peptide Phe-Gly-Phe for improving sperm motility and mucus penetration ability. The small peptide Phe-Gly-Phe comprises three amino acids: Phe, Gly and Phe.

[0011] The present invention also provides an application of the small peptide Phe-Gly-Phe in the preparation of a drug for treating asthenospermia for improving sperm function.

[0012] Preferably, the therapeutic agent is capable of modulating overall sperm motility.

[0013] Preferably, the therapeutic drug is capable of modulating the progressive motility of sperm.

[0014] Preferably, the therapeutic drug is capable of modulating the progressive motility of sperm.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] The present invention improves the function of sperm in the process of in vitro fertilization by utilizing Phe-Gly-Phe, so as to increase the fertilization success rate through artificial insemination, in vitro fertilization-embryo transfer or intracytoplasmic sperm injection, treat infertility, and greatly improve the patient's reproductive outcome. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a diagram showing the experimental results of FGF improving the sperm climbing ability in Example 1 of the present invention;

[0018] Figure 2 This is a diagram showing the experimental results of FGF improving sperm motility and increasing sperm movement level in Example 2 of the present invention. DETAILED DESCRIPTION

[0019] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the accompanying drawings, so that those skilled in the art can better understand the advantages and features of the present invention, thereby making a clearer definition of the protection scope of the present invention. The embodiments described in the present invention are only a part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without making creative work are within the scope of protection of the present invention.

[0020] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0021] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0022] The formula of the HTF solution (Human tubal fluid) used in the present invention is: 93.8mM NaCl, 4.69mM KCl, 0.2mM MgSO4, 0.37mM KH2PO4, 2.04mM CaCl2, 21.4mM lactic acid, 2.78mM glucose, 4mM NaHCO3, 331.8μM Sodium pyruvate, 200mg / 50mL BSA, and the pH is adjusted to 7.4.

[0023] The HS solution (HS high-saline solution) used in the present invention has a formula of: 135mM NaCl, 5mM KCl, 1mM MgSO4, 2mM CaCl2, 20mM HEPES, 5mM glucose, 10mM lactic acid and 1mM Na-pyruvate at pH 7.4 with NaOH.

[0024] Embodiment 1:

[0025] Climbing test to detect the ability of sperm to penetrate viscous media

[0026] Take 1mL of sperm sample from a volunteer with normal parameters, add 5mL of HS and place in an incubator at 37℃, 5% CO2 for 1h. Take 3mL of the supernatant with good activity and no impurities after the upstream. Centrifuge and wash once at 300g for 6min. Resuspend the sperm pellet in 1mL of HTF and reserve 800μL of sperm sample. After 3h of capacitation in an incubator at 37℃, 5% CO2, the sperm suspension was divided into 3 groups. Add different drugs, the FGF concentration was 100μM, and 10μM progesterone P4 was used as a positive control. 5min after the incubation, insert the capillary glass tube filled with 1% methylcellulose into the sperm suspension with different drugs added, and let the sperm climb in the incubator for 1h. Finally, transfer it to a microscope with a slide as a fixed matrix for observation and statistics of the sperm density of each glass tube at 1cm, 2cm, and 3cm.

[0027] like Figure 1 As shown, 100 μM FGF can significantly enhance the sperm climbing ability at 2 cm and 3 cm.

[0028] Embodiment 2:

[0029] CASA system detects sperm motility

[0030] Sperm samples from volunteers with normal parameters were obtained after upstreaming to obtain sperm suspension with good activity and no impurities. Centrifuge at 300g for 6min, discard the supernatant, and resuspend in HTF capacitation solution. The sperm suspension was divided into 2 groups. First, 200mM FGF mother solution was prepared with DMSO, and then 10mM FGF was prepared with HS solution. Finally, FGF with a final concentration of 100μM was added to the sperm suspension. To control the variable, DMSO was treated in the same way as the control group. CASA detects sperm motility parameters, mainly recording Totalmotility, Progressive, VAP, VSL, VCL, LIN, ALH, BCF, STR, and LIN. To determine whether sperm is superactivated, the parameters need to meet VCL≥150μm / s, LIN≤50%, and ALH≥7.0μM at the same time. Sort A is the percentage of sperm superactivation, which is the ratio of superactivated sperm to the total number of sperm.

[0031] like Figure 2 As shown, 100 μM FGF can enhance the total sperm motility and forward motility.

[0032] In summary, the present invention has found that small peptides can improve the movement and function of mature human sperm under in vitro culture conditions. In vitro experiments have shown that compared with the control group, the sperm motility level was significantly improved and the ability to penetrate mucus was significantly increased after the addition of Phe-Gly-Phe. The application of Phe-Gly-Phe can effectively improve the function of sperm during in vitro fertilization by enhancing sperm motility and improving motility, so that the sperm after the optimization or treatment of the present invention can increase the fertilization success rate through artificial insemination, in vitro fertilization-embryo transfer or intracytoplasmic sperm injection, treat infertility, and greatly improve the patient's reproductive outcome.

[0033] The description and practice disclosed in the present invention are easy to think and understand for ordinary technicians in the technical field, and several improvements and modifications can be made without departing from the principles of the present invention. Therefore, modifications or improvements made without departing from the spirit of the present invention should also be regarded as the protection scope of the present invention.

Claims

1. A small peptide Phe-Gly-Phe for improving sperm motility and mucus penetration, characterized in that: The small peptide Phe-Gly-Phe comprises three amino acids: Phe, Gly and Phe.

2. Use of the small peptide Phe-Gly-Phe according to claim 1 in the preparation of a drug for treating asthenospermia for improving sperm function.

3. The use according to claim 2, characterized in that: The therapeutic drug is capable of regulating the overall motility of sperm.

4. The use according to claim 2, characterized in that: The therapeutic drug is capable of regulating the forward motility of sperm.

5. The use according to claim 2, characterized in that: The therapeutic drug can enhance the ability of sperm to penetrate mucus.

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