阅读说明：本技术 一种5-Methylcytidine在制备促生精药物中的应用 (Application of 5-Methylcytidine in preparation of spermatogenic promoting drugs ) 是由 唐冲 张滢 于 2021-01-12 设计创作，主要内容包括：本发明公开一种5-Methylcytidine在制备促生精药物中的应用,5-Methylcytidine在治疗无精症或少精症具有独特的疗效。(The invention discloses application of 5-Methylcytidine in preparing a spermatogenic promoting medicine, and the 5-Methylcytidine has unique curative effect on azoospermia or oligospermia.)

1. An application of 5-Methylcytidine in preparing the medicine for promoting the generation of sperm is disclosed.

2. The use of 5-Methylcytidine in the preparation of a spermatogenic medicament according to claim 1, wherein the spermatogenic medicament is a medicament for the treatment of azoospermia.

3. The use of 5-Methylcytidine in the preparation of a spermatogenic medicament according to claim 1, wherein the spermatogenic medicament is a medicament for the treatment of oligospermia.

4. The use of 5-Methylcytidine in the preparation of a spermatogenic medicament according to claim 1, wherein the spermatogenic medicament is for promoting the spermatogenic ability of a patient with azoospermia to recover.

5. The use of 5-Methylcytidine according to claim 1 for the preparation of a spermatogenic medicament, wherein the spermatogenic medicament is a medicament for increasing the methylation level of RNA of spermatogenic cells.

6. The use of 5-Methylcytidine in the preparation of a spermatogenic medicament according to claim 1, wherein the spermatogenic medicament is a medicament for the treatment of non-obstructive azoospermia and oligoasthenospermia.

7. An application of 5-Methylcytidine in promoting the generation of sperm from testis is disclosed.

8. An application of 5-Methylcytidine in restoring sperm production from testis is disclosed.

Technical Field

The invention relates to the field of medicines, and in particular relates to application of 5-Methylcytidine in preparation of a spermatogenic promoting medicine.

Background

Infertility accounts for about 15% of the population of the child-bearing age, with male factors accounting for 50% and azoospermia accounting for 10%. Azoospermia can be classified into obstructive azoospermia and non-obstructive azoospermia, wherein obstructive azoospermia is caused by the fact that sperms cannot be normally discharged due to obstruction of epididymis or vas deferens caused by inflammation, congenital malformation or tumor compression, and the like, and the spermatogenesis of the obstructive azoospermia is not affected, so that the obstructive azoospermia is clinically curable. Non-obstructive azoospermia and oligoasthenospermia are infertility caused by the production of non-mature spermatozoa due to spermatogenesis dysfunction. Clinically, for infertility caused by non-obstructive azoospermia and oligospermia, except for a small part of infertility caused by abnormal hormone secretion, the rest azoospermia is caused by genetic factors, so that no effective treatment medicine exists, and the only solution is assisted reproduction technology, namely test-tube infants.

The spermatogenesis process refers to a process in which spermatogonia finally form mature sperms through mitosis, meiosis and deformation, and the process is cooperatively regulated by a plurality of genes, wherein dysregulation of any one of the genes can cause spermatogenesis disorder, and finally causes azoospermia or oligoasthenospermia in clinic. From the pathological typing perspective, spermatogenic disorders can be mainly classified as supportive cell only syndrome (only supportive cells in the seminiferous tubules, without spermatogenic cells), spermatogenic arrest in spermatogonial cells (meiosis), spermatogenic arrest in spermatids (dysspermia). Most commonly seen in clinic is that spermatogenesis stagnates in the sperm cell stage, i.e. the anaphase dysgenosis of sperm is unable to produce extended sperm with normal morphology. At present, no effective treatment means exists clinically aiming at the support cell syndrome only and the azoospermia with spermatogenic stagnation in spermatogonium and spermatocyte, the assisted reproduction technology can only use for supplying sperms, and the azoospermia with spermatogenic stagnation in spermatocyte can also be subjected to microscopic sperm collection operation to search possible haploid sperms for carrying out the self sperm assisted reproduction. In any way, the clinical treatment means for patients with azoospermia and oligospermia is artificial assisted reproduction, and the self spermatogenesis and natural pregnancy can not be recovered.

Disclosure of Invention

The invention aims to provide a novel spermatogenic medicament.

In order to solve the technical problem, the invention provides application of 5-Methylcytidine in preparing a spermatogenic promoting medicine.

Preferably, the spermatogenic medicament is a medicament for treating azoospermia.

Preferably, the spermatogenic medicament is a medicament for treating oligospermia.

Preferably, the spermatogenic medicament is used for promoting the spermatogenic ability of the azoospermia patient to recover.

Preferably, 5-Methylcytidine increases the level of methylation of RNA in spermatogenic cells.

Preferably, the spermatogenic promoting medicine is a medicine for treating non-obstructive azoospermia and oligospermia.

In addition, an application of 5-Methylcytidine in promoting sperms generation from testis is provided.

And, an application of 5-Methylcytidine in restoring sperm production from testis.

The structural formula of Methylcytidine is shown as follows:

the spermatogenesis arrest is caused by gene regulation abnormality in the spermatogenesis process, especially the 'delayed translation' abnormality of spermatogenesis specific expression gene. The phenomenon of "delayed translation" is a specific gene expression regulation mode in the process of spermatogenesis, and the phenomenon of uncoupling of transcription and translation is called "delayed translation" because the template RNA required for protein translation in late-stage prolonged sperm is derived from RNA stored in early-stage spermatozoa because of the phenomenon of cell nucleus condensation and the stop of transcription in the process of transformation of round sperm to prolonged sperm in the late-stage spermatogenesis, i.e. no new RNA is generated. These RNAs stored in early spermatogenic cells need to bind to the corresponding RNA-binding proteins as complexes to be stored in the subunit RNP (ribonucleoprotein) in order to keep themselves undegraded and translated in advance, and if this process is deregulated, the "delayed translation" is out of control, which in turn leads to dysspermia. The most important link of the delayed translation of RNA is the storage of RNA in RNP, the process depends on the interaction between RNA and RNA binding protein, and the interaction depends on the modification of RNA, especially methylation modification, namely, the methylation modification site on RNA can specifically recruit related RNA binding protein and form a complex to store RNA in RNP, protect RNA from degradation and translation, and reserve the delayed translation at the later stage to promote the normal process of spermatogenesis.

Therefore, the mechanism of promoting spermatogenesis by 5-Methylcytidine is:

1.5-Methylcytidine is a cytidine that can enter spermatogenic cells without hindrance and integrate into cellular RNA.

2. The RNA integrated into the spermatogenic cells can improve the methylation level of the corresponding RNA and increase RNA methylation sites, thereby promoting the RNA to be stored in the RNP, twisting RNA delayed translation abnormality and promoting the spermatogenesis to be recovered.

Drawings

FIG. 1 shows the results of sperm concentration and sperm morphology.

FIG. 2 is a photograph comparing testis size.

Fig. 3 is a testis weight comparison graph.

FIG. 4 is a staining morphology of testis tissue sections.

FIG. 5 is a 35-day reproduction comparison chart.

FIG. 6 is a graph comparing propagation at 70 days.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is obvious that the embodiments described are only some representative embodiments of the present invention, rather than all embodiments, and all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.

Example 1

Adult ICR male mice are divided into three groups, namely a control group (wild type WT), a model group (busulfan construction module) and a dosing group, wherein the control group and the model group are respectively 8, the dosing group is subjected to gradient concentration dosing, and the three concentration gradients are respectively 1mg/kg, 2mg/kg and 4mg/kg, and each group is 8. All mice in the model group and the administration group use a 35mg/kg busulfan gastric lavage and model azoospermia model. After 40 days of molding, the azoospermia phenotype reaches a peak, the spermatids in the testicular seminiferous tubules are greatly reduced, no mature sperms are generated, and the mice are sterile. Beginning on the 40 th day of modeling, the administration group starts to inject 5-Methylcytidine into the abdominal cavity, the dosages are respectively 1mg/kg, 2mg/kg and 4mg/kg which are dissolved in PBS solution, and the administration is continuously carried out for 35 days (one spermatogenic period) by the intraperitoneal injection every day; model groups PBS solutions were given under the same conditions and frequency. After 35 days, 6 mouse epididymal tail sperms are taken from each group for sperm motility analysis, and mouse testicles are taken for morphological analysis; 2 mice were left in each group for mating experiments with wild type females. The results are as follows:

1. as shown in fig. 1, sperm analysis results: compared with the sperm parameters of the wild mice in the control group, the sperm density and the motility of the model group are obviously reduced; although the sperm density and the sperm motility of the three administration concentration groups are not restored to the level of the control group, the sperm density and the motility are obviously increased compared with the model group, and the sperm parameters of the three administration concentration groups have no obvious difference.

2. As shown in fig. 2 and 3, testis size and weight results: the average mass of the testes of the control group mice is 138mg, the testes of the model group are obviously reduced, and the average mass is 31 mg; the testis size of the administration group is remarkably recovered compared with that of the model group, wherein the average number of the testis masses of the 1mg/kg concentration administration group is 48mg, the average number of the testis masses of the 2mg/kg concentration administration group is 44mg, the average number of the testis masses of the 4mg/kg concentration administration group is 51mg, and no obvious difference exists among the three concentration groups.

3. As shown in fig. 4, the result of morphological analysis of stained testis tissue sections: compared with the control group, the sperms in the testis spermatid tubules of the model group are seriously damaged and are vacuolated, the spermatid cells are rarely seen, and no mature spermatid cells exist; seminiferous complete recovery of most seminiferous tubules of testis in three concentration administration groups, the seminiferous sperm cells in the lumen are full, mature sperm cells can be seen, and the seminiferous sperm cells are recovered in all three concentration groups without obvious difference.

4. As shown in fig. 5 and 6, the mating experiment results: after the model group and the administration group are subjected to Busulfan treatment for 40 days, namely, the azoospermia reaches the peak, two male mice and two female mice are respectively selected from the control group, the model group and the three concentration administration groups and are respectively combined in a cage, and two sperm producing periods (70 days) are observed in the breeding condition. We recorded the litter size of each mating cage at 35 and 70 days, respectively, and the spermatogenesis recovery was significant for the dosed group compared to the model group.

Various modifications may be made to the above without departing from the spirit and scope of the invention as defined by the appended claims. The scope of the invention is therefore intended to be limited not by the above description, but rather by the scope of the appended claims. The invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and the scope of protection is still within the scope of the invention.