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Marker protein related to asthenospermia and application of marker protein

A technology for asthenozoospermia and histones, applied in the field of biomedicine, can solve problems such as unclear molecular mechanism

Pending Publication Date: 2021-05-07
上海市生物医药技术研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main causes of low sperm motility include abnormal metabolism of the testis or epididymis, defects in the structure of the sperm tail, and necrospermia, but the molecular mechanism is still unclear

Method used

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  • Marker protein related to asthenospermia and application of marker protein
  • Marker protein related to asthenospermia and application of marker protein
  • Marker protein related to asthenospermia and application of marker protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0135] Example 1 Research on Thiothiolated Proteome in Human Sperm

[0136] 1.1 Materials and methods

[0137] 1.1.1 Experimental materials

[0138] 1.1.1.1 Experimental animals

[0139] Adult male C57BL / 6 mice (8 weeks old) were purchased from Shanghai Slack Experimental Animal Co., Ltd. (SCXK (Shanghai) 2017-0005) and Shanghai Xipuer-Bikai Experimental Animal Co., Ltd. (SCXK (Shanghai) 2013-0016) , AB+ zebrafish used in experiments were purchased from Beijing Weitong Lihua Experimental Animal Company (SCXK (Beijing) 2014-0001). The mice were kept in the SPF-level experimental animal room of the Shanghai Institute of Family Planning Science (the license for the use of experimental animals: SYXK (Shanghai) 2013-0027), and the feeding conditions were as follows: temperature 24°C, relative humidity 50%, daily light 14hr, free Diet and water. The zebrafish were kept in the Fish Experimental Animal Room of the Shanghai Family Planning Research Institute under constant temperat...

Embodiment 2

[0197] Example 2 Relationship between protein sulfhydrylation and sperm motility

[0198] The research of the present invention finds that the sulfhydryl modification of sperm protein is involved in the regulation of the motility of sperm, and this embodiment will mainly focus on this hypothesis.

[0199] 2.1 Experimental materials

[0200] 2.1.1 Experimental materials and reagents

[0201] The sperm used in the experiment of this example is from the same source as that used in the above example, and other reagents are also the same as those described above.

[0202] 2.1.2 Antibodies

[0203] The H3K4Me3, H3K9Me3 and H3K27Me3 antibodies used in WB detection were purchased from Abcam. The article numbers are ab8580, ab8898 and ab4729 respectively.

[0204] 2.2 Experimental method

[0205] 2.2.1 Separation of sperm with different motility by upstream sperm method

[0206] The sperm upstream method is a simple sperm sorting method that can separate the motile sperm from a s...

Embodiment 3

[0228] Example 3 Determination of Histone H3 Thiosulfhydryl Modification Sites

[0229] Previous studies have confirmed that the level of sulfhydrylation of H3 protein in asthenospermia sperm significantly decreased, and the level of H3 sulfhydrylation was positively correlated with sperm motility. In this example, the modification site of sulfhydrylation of histone H3 is determined.

[0230] 3.1 Experimental materials

[0231] 3.1.1 Cell lines

[0232] Mouse Leydig cell line TM-3 was purchased from Suzhou Beina Chuanglian Biotechnology Co., Ltd., the product number is BNCC100249. The mouse spermatogonial stem cell line C18-4 was donated by Professor He Zuping's laboratory at Shanghai Jiao Tong University School of Medicine. Cells were cultured in DMEM / F12 medium containing 10% fetal bovine serum (FBS) at 37 °C with 5% CO 2 in a constant temperature incubator. Replace with fresh culture medium every 2-3 days. Cells for identification were cultured in 6-well plates, and v...

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Abstract

The invention provides a marker protein related to asthenospermia and an application of the marker protein, and particularly discovers that sulfur-sulfhydrylation modification of H3 protein in sperm protein participates to regulation and control of establishment and maintenance of sperm motility, and abnormal sulfur-sulfhydrylation modification of H3 participates in a pathological process of asthenospermia. The sperm motility can be obviously improved by improving the H3 sulfhydrylation level, so that the asthenozoospermia is prevented or treated.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular, the invention relates to asthenospermia-associated marker protein and its application. Background technique [0002] Infertility is the inability to conceive after unprotected intercourse with the same partner at an appropriate frequency for one year or more. At present, there are more than 40 million infertile patients in China, accounting for 12.5% ​​of China's total childbearing population. About half of infertility is caused by the man. With the number of male infertility patients increasing year by year, male infertility has become one of the major problems in clinical medicine worldwide. [0003] Asthenozoospermia (AS) is one of the main causes of male infertility, and about 17.5% of male infertile patients have asthenozoospermia. Asthenospermia refers to sperm motility lower than normal sperm motility, usually defined as less than 50% motile sperm or less than 25% forward motil...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K38/06A23L33/10C07K14/47G01N33/68A61P15/08A61P15/00
CPCA61K45/00A61K38/063C07K14/47G01N33/689A61P15/08A61P15/00A23L33/10G01N2800/367G01N2333/47A23V2002/00A23V2200/30A23V2250/31
Inventor 李润生祁琪
Owner 上海市生物医药技术研究院
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