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Application of mir-1288 in diagnosis or treatment of osteoarthritis diseases

A technology of mir-1288, 1.mir-1288, applied in the application field of miR-1288 in the diagnosis or treatment of osteoarthritis disease, can solve the problem of no treatment method and the like

Active Publication Date: 2020-02-21
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the irreversibility of cartilage tissue lesions, an effective treatment method has not yet appeared clinically

Method used

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  • Application of mir-1288 in diagnosis or treatment of osteoarthritis diseases
  • Application of mir-1288 in diagnosis or treatment of osteoarthritis diseases
  • Application of mir-1288 in diagnosis or treatment of osteoarthritis diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 High-throughput sequencing

[0035] 1. Sampling

[0036] From October 2012 to December 2015, patients with osteoarthritis who were treated in the Department of Orthopedics of Peking Union Medical College Hospital were collected. A total of 10 cases were collected as the case group. A total of 4 cases were collected from patients with other diseases who were hospitalized in the Department of Orthopedics during the same period as controls. The synovial fluid samples of all subjects were obtained, numbered and stored in a -80°C low-temperature refrigerator.

[0037] The patients in the case group were all in line with the diagnostic criteria of knee OA and underwent knee arthroplasty; the control group were patients with meniscus injury and cruciate ligament injury who underwent arthroscopic surgery. According to the 1995 American Rheumatology Association diagnostic criteria for osteoarthritis, a patient with severe osteoarthritis and an indication for knee repl...

Embodiment 2

[0060] Example 2 RT-PCR verification of the expression of miR-1288 and SHISA4 in patients with osteoarthritis

[0061] 1. Materials

[0062] The synovial fluid of 36 patients with osteoarthritis and the synovial fluid of 10 controls were selected and grouped and numbered. The patients in the case group were all in line with the diagnostic criteria of knee OA and underwent knee arthroplasty; the control group were patients with meniscus injury and cruciate ligament injury who underwent arthroscopic surgery.

[0063] 2. Method

[0064] 2.1 Carry out total RNA extraction to synovial fluid, with the extraction method of embodiment 1.

[0065] 2.2 Reverse transcription

[0066] (1) mRNA reverse transcription

[0067] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0068] Using a reverse transcription ...

Embodiment 3

[0087] Example 3 Verification of the relationship between miR-1288 and SHISA4 target genes

[0088] 1. Design and synthesis of antisense oligonucleotides against miR-1288 (anti-miR-1288)

[0089] According to the sequence information of miR-1288, the antisense oligonucleotide sequence and random control sequence of miR-1288 were designed and synthesized by Dalian Bao Biotechnology Co., Ltd. The antisense oligonucleotide sequence of miR-1288 is shown as SEQ ID NO.4 shown.

[0090] 2. Cell culture

[0091]Wash the sterile knee articular cartilage tissue removed from the joint replacement with phosphate buffered saline (petal bovineserum, PBS) containing penicillin (100 U / mL) and streptomycin (100 μg / mL) and place it in a sterile petri dish for several times , trim the cartilage to a size of 1mm×1mm×1mm; add 3 times the volume of 0.25% trypsin to digest at 37°C for 30-50 minutes, then gently blow and beat to discard the supernatant, and rinse with D-Hanks (Baihao Biological) so...

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Abstract

The invention discloses an application of miR-1288 to preparation of a medicine for treating SHISA4 gene-related diseases, and a kit for diagnosing early osteoarthritis. The kit comprises a reagent for detecting the expression level of the miR-1288. The invention further discloses a medicine for treating the osteoarthritis. The medicine comprises a miR-1288 inhibitor. By the aid of miRNA for detecting the osteoarthritis, early detection can be rapidly and effectively realized, and therapeutic targets and important evidence are provided for clinical applications such as gene therapy and medicine therapy.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of miR-1288 in diagnosing or treating osteoarthritis diseases. Background technique [0002] Osteoarthritis (OA) is a chronic progressive disease characterized by the degeneration, destruction, disappearance of articular cartilage and bone hyperplasia around the joints. The prevalence increases significantly with age. The main clinical manifestations are: pain, swelling, deformity, and limited movement of the affected joints. The main X-ray findings are the narrowing of the joint space of the affected joints, subchondral bone sclerosis, and formation of surrounding osteophytes. At present, the pathogenesis of OA is still unclear, and a more unified explanation is that mechanical and biological factors lead to the degeneration of chondrocytes, extracellular matrix and subchondral bone. [0003] Articular cartilage is mainly composed of chondrocytes, type II ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/7105A61P19/02C12Q1/6883
CPCA61K31/7105C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 范彧翁习生林进杨波叶伟亮
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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