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Application of CTRP3 protein (C1qTNF related protein 3)

A technology of protein and follicle-stimulating hormone, which is applied in the fields of biotechnology and medicine, can solve problems such as decreased ovulation, increased abnormal embryos, follicle atresia, etc., and achieves the effect of enhancing the effect and enhancing the sensitivity

Active Publication Date: 2016-12-07
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that the number of developing follicles in PCOS patients after FSH stimulation treatment is more than that of normal people, but due to the excessive production of estrogen, a negative feedback regulation is formed, which inhibits the level of FSH in the body, so that a large number of developing follicles are stimulated by FSH. Insufficient resulting in follicular atresia and reduced ovulation
Increasing the dose of exogenous FSH in this process is also a routine treatment strategy for PCOS, but the use of high doses of FSH will increase the probability of abnormal embryos
There is no research report on the regulation of CTRP3 on follicle development

Method used

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  • Application of CTRP3 protein (C1qTNF related protein 3)
  • Application of CTRP3 protein (C1qTNF related protein 3)
  • Application of CTRP3 protein (C1qTNF related protein 3)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Immunohistochemical analysis of the expression distribution of CTRP3 protein in mouse ovarian tissue.

[0035] Take 7-week-old normal female mice, kill them by cervical dislocation, separate the ovarian tissue, fix the ovarian tissue with 4% paraformaldehyde for 3 hours, and then make paraffin-embedded sections. Wash 3 times, 10 minutes each time. 70% ethanol dehydration for 45 minutes, 85% ethanol dehydration for 45 minutes, 95% ethanol dehydration for 45 minutes, 100% ethanol dehydration for 45 minutes, and 100% ethanol dehydration for 45 minutes. 1 / 2 ethanol + 1 / 2 xylene transparent for 30 minutes, xylene transparent for 5 minutes, xylene transparent for 5 minutes, observe the tissue block until transparent. Permeate the wax at 62°C for 120 minutes. Then put the tissue block into the mold, place it in a good position, pour it with paraffin for embedding, cool it on the cold table of the paraffin embedding machine, and store it at 4°C for later use. Slic...

Embodiment 2

[0037] Example 2: Immunofluorescence analysis of the expression distribution of CTRP3 protein in human ovarian tissue.

[0038] Human ovarian tissue samples were voluntarily donated by patients undergoing ovariectomy for cervical cancer in the Department of Obstetrics and Gynecology of the First Affiliated Hospital of Jinan University with informed consent. The operations of fixing, embedding, slicing and incubating the primary antibody of the removed human ovarian tissue were the same as the immunofluorescence steps in Example 1. After the primary antibody stayed overnight, Alexa Fluor 594-labeled goat anti-rabbit secondary antibody was added dropwise, and incubated in a humid box in the dark for 60 minutes. Rinse slowly with PBS for 5 minutes x 3 times, stain with DAPI (Wuhan Boster) and protect from light for 10 minutes. Wash with PBS 3 times, 2 minutes each time. Anti-fluorescent quencher (Wuhan Boster) was added dropwise, and the slides were sealed and photographed unde...

Embodiment 3

[0039] Example 3: Effect of CTRP3 protein on follicle development of ovaries cultured in vitro

[0040] Schematic diagram of isolated ovarian tissue culture, such as image 3 As shown in A. The ovaries of 10-day-old and 12-day-old Kunming female mice were carefully separated under a stereoscope to avoid damage. Place two paired ovaries of each mouse in two Millipore culture chambers with 0.4 μm pores, place the culture chambers in a 24-well plate supplemented with 400 μL of ovarian tissue culture medium, and store at 37°C, 5% CO 2 Cell culture incubator under the conditions. The composition of ovarian tissue culture medium is: DMEM / F12 basal medium (GIBCO) + 0.1% bovine serum albumin by mass + 0.1% advanced bovine serum albumin Albumax II (GIBCO) + 1:20 diluted ITS (GIBCO, 100×, insulin-transferrin-selenium)+0.5 mg / mL L-ascorbic acid (Sigma)+1:100 diluted penicillin-streptomycin solution (penicilin-streptomycin, GIBCO, 100×). At the beginning of the culture, different conc...

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Abstract

The invention discloses an application of CTRP3 protein (C1qTNF related protein 3), and belongs to the fields of biotechnology and medicines. The CTRP3 protein provided by the invention can enhance the sensitivity of follicle to follicle stimulating hormone; and by identifying the expression of the CTRP3 protein in human and mouse ovaries, it indicates that the CTRP3 protein is autologous protein of the ovaries; therefore, the the CTRP3 protein can be safely developed and utilized as a drug. The effect of the CTRP3, as a follicle stimulating hormone sensitizer, on promoting follicle development is verified from three different levels, namely isolated ovarian tissue culture, isolated preantral follicle culture and living mesovarium injection, indicating that the CTRP3 protein, as the follicle stimulating hormone sensitizer, is effective on promoting preantral follicle and antral follicle development. The invention provides the effect and the mechanism of the CTRP3 participating in follicle development regulation and indicates that the CTRP3 protein, as the follicle stimulating hormone sensitizer, is applicable to the treatment of female ovarian hypoplasia diseases, so that the effect of the follicle stimulating hormone on the development of the follicle is enhanced.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine, and particularly relates to the application of a CTRP3 protein as a follicle stimulating hormone sensitizer. Background technique [0002] In women's normal menstrual cycle, there are generally about 20 preantral follicles, but the secretion of estradiol is extremely low. In the late stage of follicle development in the menstrual cycle, only one mature follicle develops to ovulate and secretes a high level of estradiol at the same time. Estradiol stimulates endometrial hyperplasia. In women with normal menstrual cycles, the use of exogenous hormones such as follicle stimulating hormone (FSH) can induce the growth of about 15 to 30 preovulatory follicles. However, in some patients, the level of endogenous gonadotropin increases, and there are follicles in the ovary, but the response to exogenous gonadotropin stimulation is low, and only 2 to 3 preovulatory follicles can be obtained afte...

Claims

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Application Information

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IPC IPC(8): A61K38/19A61K38/24A61P15/08
CPCA61K38/191A61K38/24A61K2300/00
Inventor 禹艳红卢小圣毛周飞杨柳红齐绪峰蔡冬青冯珊珊肖銮娟
Owner JINAN UNIVERSITY
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