Applicationof let-7c/let-7d in intrauterine adhesion and/or thin endometriumdiagnosis and treatment

A technology of let-7d and intrauterine adhesions, applied in the direction of medical preparations containing active ingredients, measurement/testing of microorganisms, biochemical equipment and methods, etc., to achieve the effect of increasing the number of glands and the thickness of the endometrium

Inactive Publication Date: 2019-04-16
NANJING DRUM TOWER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, the role of let7 in the endometrium has not been reported

Method used

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  • Applicationof let-7c/let-7d in intrauterine adhesion and/or thin endometriumdiagnosis and treatment
  • Applicationof let-7c/let-7d in intrauterine adhesion and/or thin endometriumdiagnosis and treatment
  • Applicationof let-7c/let-7d in intrauterine adhesion and/or thin endometriumdiagnosis and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Expression of let-7a, let-7b, let-7c, let-7d, let-7e, let-7f and let-7g in endometrial tissues of patients with normal and severe intrauterine adhesions

[0024] 1. Materials, reagents, equipment

[0025] 1.1 Source of human endometrial tissue

[0026] Collect 11 cases of normal endometrial tissue and 11 cases of endometrial tissue from patients with severe intrauterine adhesion syndrome. When obtaining endometrial tissue, the follicle size was detected by ultrasound to ensure that the endometrial tissue acquisition stage was 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.

[0027] 1.2 Main reagents

[0028] Trizol (Invitrogen), reverse transcriptase (Takara), Qpcr enzyme (Roche), let-7 family member reverse transcription and qPCR primers.

[0029] 1.3 Main instruments

[0030] qPCR instrument (Roche), PCR instrument (ABI)

[0...

Embodiment 2

[0036] Example 2 Changes of fibrosis markers in endometrial epithelial cells after let-7c / let-7d reduction.

[0037] 1. Materials, reagents, equipment

[0038] 1.1 Main reagents

[0039] Collagenase (Sigma), Hyaluronidase (Sigma), DNAase (Roche), Epithelial Cell Culture Medium (Gibco), Serum (Gibco), Trizol (Invitrogen), Reverse Transcriptase (Takara), QPCR Enzyme (Roche), let-7c / 7dinhibitor (Ribobio, Guangzhou).

[0040] 1.2 Main instruments

[0041] Cell incubator, shaker, qPCR instrument (Roche), PCR instrument (ABI)

[0042] 1.3 Main method

[0043] 1.3.1 Isolation and culture of primary endometrial epithelial cells

[0044] 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 diges...

Embodiment 3

[0049] Example 3 After let-7c / let-7d intervene in a mouse model of endometrial fibrosis, endometrial fibrosis changes.

[0050] 1. Materials, reagents, equipment

[0051] 1.1 Main reagents

[0052] HE staining solution (Solarbio), Masson staining solution (Thermo), angomir let-7c / let-7d and angomir control (Guangzhou Ruibo).

[0053] 1.2 Main instruments

[0054] Tissue embedding machine, slicer

[0055] 1.3 Main method

[0056] 1.3.1 Establishment of mouse endometrial fibrosis model

[0057] Select BALB / c female mice of 8-10 weeks old, weighing about 20 g, in estrus. The mice were anesthetized by intraperitoneal injection of 0.2ml of 4% chloral hydrate, and the limbs were fixed with medical adhesive tape. Cut the abdomen with surgical scissors, insert a self-made scraper into the upper part of the uterus, and scrape back and forth on one side of the uterus for about 2 minutes, resulting in rough endometrial surface; Intrauterine injection of 20 μl 2.5mg / ml LPS with insu...

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Abstract

The invention discloses applicationof let-7c/let-7d in intrauterine adhesion and/or thin endometriumdiagnosis and treatment. It is discovered that for an intrauterine adhesion patient, let-7c/let-7d expression in the endometrium is remarkably reduced. In-vitro experiments show that after the let-7c/let-7d is reduced, fibroses endometrial epithelium cells are increased. In-vivoanimal experiments show that by supplementing the let-7c/let-7d, the endometriumfibrosis degree of an endometriumfibrosis mouse is remarkably reduced. Therefore, the let-7c/let-7d possiblyplay an importanteffect in the aspects of intrauterine adhesion and/or thin endometriumdiagnosis and treatment, and can be used for preparing medicine for treating intrauterine adhesion and/or thin endometriumdiagnosis and treatmentor preparing medicine for treating other diseases related to uterusfibrosis.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals and relates to the application of let-7c / let-7d in the diagnosis and treatment of 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 method is surgical separation of adhesions, and placement of intrauterine devices or balloons or biomaterials to block the anterior and posterior walls of the uterus to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61P15/00A61P15/08A61P15/06C12Q1/6883
CPCA61K45/00A61P15/00A61P15/06A61P15/08C12Q1/6883C12Q2600/158
Inventor 胡娅莉赵光锋宋敏敏李若天
Owner NANJING DRUM TOWER HOSPITAL
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