Application of miR-21 in preparation of medicine for treating intrauterine adhesion and/or thin inner membranes

A technology of intrauterine adhesions and mir-21, applied in the application field of medicine, can solve problems affecting endometrial fibrosis and other problems

Active Publication Date: 2017-12-29
NANJING DRUM TOWER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether there is a correlation between miR-21 and intrauterine adhesion syndrome, and whether miR-21 affects endometrial fibrosis has not been reported.

Method used

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  • Application of miR-21 in preparation of medicine for treating intrauterine adhesion and/or thin inner membranes
  • Application of miR-21 in preparation of medicine for treating intrauterine adhesion and/or thin inner membranes
  • Application of miR-21 in preparation of medicine for treating intrauterine adhesion and/or thin inner membranes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Expression of miR-21 in endometrial tissues of patients with normal and severe intrauterine adhesions

[0028] 1. Materials, reagents, equipment

[0029] 1.1 Source of human endometrial tissue

[0030] Collect 5 cases of normal endometrial tissues and 5 cases of endometrial tissues from patients with severe intrauterine adhesion syndrome. When obtaining endometrial tissues, the size of follicles was detected by ultrasound to ensure that all the stages of endometrial tissue acquisition were unified into the late stage of proliferation. All participants signed a written informed consent form, which was approved by the Ethics Committee of Nanjing Drum Tower Hospital.

[0031] 1.2 Main reagents

[0032] MicroRNA in situ detection probe HAS Probe TM (Pengji Kaifeng Biotechnology Co., Ltd.), hybridization kit (Pengji Kaifeng Biotechnology Co., Ltd.), anti-digoxigenin antibody (Anti-Digoxigenin-AP, Roche), NBT / BCIP color development (Pengji Kaifeng Biotechnology C...

Embodiment 2

[0039] Example 2 Effect of miR-21 on fibrosis of endometrial epithelial cells

[0040] 1. Materials, reagents, equipment

[0041] 1.1 Main reagents

[0042] Collagenase (Sigma), Hyaluronidase (Sigma), DNAase (Roche), Epithelial Cell Culture Medium (Gibco), Serum (Gibco), Trizol (Invitrogen), Reverse Transcriptase (Takara), Qpcr Enzyme (Roche).

[0043] 1.2 Main instruments

[0044] Cell incubator, shaker, qPCR instrument (Roche), PCR instrument (ABI)

[0045] 1.3 Main method

[0046] 1.3.1 Isolation and culture of primary endometrial epithelial cells

[0047] Place the endometrial tissue in a new 60mm petri dish, cut the tissue until no lumps are visible to the naked eye, add the prepared digestive solution, blow and mix, and place it in a 37°C incubator for 5 minutes; take out the petri dish and observe the tissue under a microscope Digestion: Add DNase at a concentration of 4mg / mL, continue to digest for 5mim; pipette the digested tissue, and drop the tissue suspension in...

Embodiment 3

[0052] Example 3 miR-21 reverses Np63-induced endometrial epithelial cell fibrosis.

[0053] 1. Materials, reagents, equipment

[0054] 1.1 Main reagents

[0055] E-cadherin antibody (Abcam), N-cadherin antibody (Abcam), fluorescent secondary antibody (Jackson), dapi-containing mounting medium (Abcam), PFA, antibody diluent (Gibco), methanol, Tween, PBS.

[0056] 1.2 Main instruments

[0057] Fluorescence microscope (Leica), shaker

[0058] 1.3 Main method

[0059] The expression of E-cadherin and N-cadherin was detected by cell immunofluorescence. After the endometrial epithelial cells were sliced, the Np63 adenovirus infected the cells for 24 hours. After 48 hours of transfection with miR-21, the slides loaded with cells were taken out. Wash 3 times with 1×PBS, 5 min each time, then fix with PFA at room temperature for 15 min, wash 3 times with 1×PBS, 5 min each time. Treat the cells with pre-cooled methanol for 5 minutes, wash with 1×PBS 3 times, 5 minutes each time....

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Abstract

The invention discloses application of miR-21 in preparation of medicine for treating intrauterine adhesion and / or thin inner membranes. The study finds that the miR-21 expression for endometrial epithelium cells and mesenchymal cells is obviously reduced, and particularly the decrease for epithelial cells is most remarkable. Moreover, in vitro results indicate that the EMT of the endometrial epithelium cells is inverted through miR-21, and fibrosis is inhibited. Therefore, the miR-21 possibly plays important roles in diagnose and treatment aspects of intrauterine adhesion and / or thin inner membranes, and can be used for preparing medicine for diagnosing and treating intrauterine adhesion and / or thin inner membranes or preparing medicine for treating other diseases related to womb fibrosis.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals and relates to the application of miR-21 in the preparation of medicines for treating intrauterine adhesion and / or thin endometrium. Background technique [0002] Endometrial regenerative disorders mainly include intrauterine adhesions and thin endometrium. Intrauterine adhesions are caused by damage to the basal layer of the endometrium and dysfunction of the regeneration and repair of the functional layer, resulting in the formation of intramural adhesions characterized by endometrial fibrosis. Up to 25-30%. In my country, with the increase in the number of intrauterine surgery, especially painless abortion, the incidence of intrauterine adhesions and endometrial fibrosis has increased significantly. At present, the main treatment is surgical separation of adhesions, and placement of intrauterine devices, balloons, or biomaterials to block the anterior and posterior walls of the uterus to pre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7105A61P41/00A61P15/00C12Q1/68
CPCA61K31/7105C12Q1/6883C12Q2600/178C12Q2600/158A61P41/00A61P15/00A61K49/0054C12N15/113
Inventor 胡娅莉周艳赵光锋李若天
Owner NANJING DRUM TOWER HOSPITAL
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