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Application of micro RNA in preventing or treating femoral head necrosis caused by glucocorticoid

A technology for glucocorticoid and femoral head necrosis, which is applied in the fields of biomolecules and medicine, and can solve problems such as poor treatment effect and difficult early diagnosis

Inactive Publication Date: 2017-05-24
边焱焱
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Avascular necrosis of the femoral head (ANFH) is a common orthopedic disease with frequent occurrence, difficult early diagnosis and poor treatment effect

Method used

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  • Application of micro RNA in preventing or treating femoral head necrosis caused by glucocorticoid
  • Application of micro RNA in preventing or treating femoral head necrosis caused by glucocorticoid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 High-throughput sequencing screening of differentially expressed genes

[0039] 1. Sampling

[0040] Bone marrow tissue samples were collected from 25 patients with femoral head necrosis undergoing total hip replacement surgery. The samples were collected from patients who visited the Department of Orthopedics of Peking Union Medical College Hospital from October 2012 to December 2015. All patients were diagnosed with long-term or high-dose Application of glucocorticoids induced avascular necrosis of the femoral head. The control samples were collected from 22 hip joint bone marrow tissue samples from patients with comminuted hip joint fractures who were hospitalized in the Department of Orthopedics at the same time. The samples of all research subjects were collected, numbered and stored in a -80°C low-temperature refrigerator. All clinical samples in this study were informed to the patients and approved by the Hospital Ethics Committee.

[0041] 2. Total ...

Embodiment 2

[0052] Example 2 RT-PCR detection of has-miR-16 expression in cells treated with different concentrations of dexamethasone

[0053] 1. Sampling

[0054] Bone marrow samples were collected from 3 patients undergoing total hip replacement surgery. The samples were collected from patients with hip osteoarthritis who visited the Department of Orthopedics of Peking Union Medical College Hospital from October 2012 to December 2015. The samples of all research subjects were collected, numbered and stored in a -80°C low-temperature refrigerator. All clinical samples in this study were informed to the patients and approved by the Hospital Ethics Committee.

[0055] 2. Isolation and culture of bone marrow mesenchymal stem cells

[0056] (1) Use a sterile bone marrow puncture needle and syringe to extract bone marrow tissue from the femoral side bone.

[0057] (2) Transfer the extracted bone marrow tissue into a test tube containing culture medium and heparin anticoagulant (4000T / ml),...

Embodiment 3

[0074] Example 3 Inhibition of miR-16-5p expression

[0075] 1. Source of cells

[0076] Bone marrow mesenchymal stem cells cultured in Example 2.

[0077] 2. Design and synthesis of antisense oligonucleotides against miR-16-5p (anti-miR-16-5p)

[0078] According to the sequence information of miR-16-5p, the antisense oligonucleotide sequence of miR-16-5p and random control sequence were designed and synthesized by Dalian Bao Biotechnology Co., Ltd., and the antisense oligonucleotide sequence of miR-16-5p It is SEQ ID NO.4.

[0079] 3. Transfection

[0080] The cationic liposome method was used for transient transfection, and the operation was carried out according to the instructions of LipofectamineTM2000Transfection Reagent reagent. 24 hours before transfection, cells in good growth state were inoculated into a 6-well plate, and the cell count was about 5×104. The test was performed when the cell confluence was 50-60% after routine culture until the day of transfection....

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Abstract

The invention discloses application of micro RNA in preventing or treating femoral head necrosis caused by glucocorticoid. The invention discloses an early diagnostic kit for the femoral head necrosis caused by the glucocorticoid, and the kit comprises a reagent used for detecting the expression level of miR-16-5p. The invention further discloses medicine for treating the femoral head necrosis caused by the glucocorticoid, and the medicine contains an miR-16-5p inhibitor. When the miRNA is utilized to detect the femoral head necrosis, early detection can be quickly and effectively obtained; furthermore, a therapeutic target and an important basis are also provided for clinical application of gene therapy, drug therapy and the like.

Description

technical field [0001] The invention relates to the fields of biomolecules and medicine, in particular to the application of microRNA related to femoral head necrosis as a molecular marker in the prevention or treatment of femoral head necrosis caused by glucocorticoids. Background technique [0002] Femoral head necrosis is often called avascular necrosis of femoral head or aseptic necrosis of femoral head because its main pathology is the blockage of blood supply to the femoral head and the destruction of the head bone caused by ischemia. Avascular necrosis of the femoral head (ANFH) is a common orthopedic disease with frequent occurrence, difficult early diagnosis and poor treatment effect. Avascular necrosis of the femoral head caused by long-term high-dose steroid use has become a common clinical drug-induced disease. According to domestic reports, the incidence rate of femoral head necrosis caused by low-dose long-term application of glucocorticoids is 20%, and the in...

Claims

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Application Information

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IPC IPC(8): C12Q1/68A61K45/00A61K31/7088A61P19/08
CPCC12Q1/6883A61K31/7088A61K45/00C12Q2600/158C12Q2600/178
Inventor 边焱焱翁习生钱文伟吴志宏李红凌王兴山彭慧明
Owner 边焱焱
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