Diagnosis and treatment marker for postmenopausal osteoporosis

An osteoporosis and drug technology, applied in the field of biomedicine

Active Publication Date: 2019-03-08
GUAN BOJIAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Searching for miRNA markers related to postmenopausal osteoporosis has important clinical diagnostic and therapeutic value. At present, there are few studies on the differential expression of miRNA in postmenopausal osteoporosis, and an effective molecular marker for postmenopausal osteoporosis is provided. It is of great significance to realize the accurate diagnosis and treatment of diseases

Method used

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  • Diagnosis and treatment marker for postmenopausal osteoporosis
  • Diagnosis and treatment marker for postmenopausal osteoporosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Screening of miRNAs associated with postmenopausal osteoporosis

[0054] 1. Sample collection

[0055]Blood samples were collected from 30 healthy people and postmenopausal osteoporosis patients. The EDTA anticoagulant tube was left to stand for 10 minutes, the serum was separated by centrifugation, and stored at -20°C for later use. All the above specimens were obtained with the consent of the organizational ethics committee. Five samples were taken for detection and analysis of miRNA expression profiles, screening of differentially expressed genes, and a large sample verification experiment was carried out in all 30 samples.

[0056] Exclusion criteria: premature menopause; patients with secondary osteoporosis caused by endocrine diseases, blood diseases, connective tissue diseases, drugs and nutritional diseases; metabolic bone diseases, chronic liver and kidney diseases and other diseases that interfere with bone metabolism Patients; patients who have r...

Embodiment 2

[0085] Example 2 QPCR verification of differentially expressed miRNA-3656

[0086] 1. Large-sample QPCR verification of differentially expressed miRNA-3656.

[0087] 2. RNA extraction

[0088] Use the QIAGEN blood RNA extraction kit to extract RNA in serum, and refer to the instruction manual for specific steps.

[0089] 3. Reverse transcription:

[0090] 1) Mix 10pg-1μg total RNA template with 2μl 10× buffer, 2μl dATP (10mM), 0.5μl polyA polymerase, 0.5μl RNase inhibitor and RNase free water , the final volume was 20 μl, and incubated at 37°C for 1h.

[0091] 2) Add 1 μl of 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, and incubate at 70° C. for 5 minutes.

[0092] 3) Immediately incubate on ice for at least 2 minutes to break the secondary structure of RNA and primers.

[0093] 4) Mix the above 20 μl reaction mixture with 4 μl 5× buffer, 1 μl dNTP (10 mM), 0.5 μl M-MLV reverse transcriptase, 0.5 μl ribonuclease (RNase) inhibitor, 10 μl polyA reaction mixt...

Embodiment 3

[0109] Example 3 Effect of miRNA-3656 expression on osteogenic differentiation

[0110] Human bone marrow mesenchymal stem cells (hMSCs) were cultured, and the inhibitor of miRNA-3656 has-miR-3656mirVana TM miRNA inhibitor (US life technologies), mimic has-miR-3656 mirVana TM miRNA mimic (Life Technologies, USA) transfected hMSCs with the carrier lipofectamine RNAiMAX Reagent, and observed the effect of miRNA-3656 on the differentiation of hMSCs into osteoblast-like cells.

[0111] 1. Cell culture

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Abstract

The invention discloses a diagnosis and treatment marker for postmenopausal osteoporosis, and the marker is miRNA-3656. According to the invention, the miRNA-3656 is proved to be relevant to the occurrence and development of the postmenopausal osteoporosis through a QPCR (Quantitative Polymerase Chain Reaction); and meanwhile, the differentiation of osteoblasts is proved to be influenced by the miRNA-3656 through cell experiments, indicating that the miRNA-3656 can be taken as a molecular target to be applied to early diagnosis and treatment of the postmenopausal osteoporosis.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to markers for diagnosis and treatment of postmenopausal osteoporosis, specifically the marker is miRNA-3656. Background technique [0002] Postmenopausal osteoporosis (postmenopausal osteoporosis, PMOP) is a kind of systemic bone metabolic disease caused by estrogen deficiency, characterized by weakened bone strength, and can induce an increased risk of fracture. The coupled dynamic balance between bone formation and bone resorption is the basis for maintaining the morphological stability of the bone itself. When the coupled dynamic balance of bone formation-bone resorption is broken, bone homeostasis will be disrupted (Ensrud KE, Crandall CJ.Osteoporosis.Annals of internal medicine 2017;167(3}:Itc17-itc32.).With the gradual increase of age or the rapid decline of hormone levels after menopause, and other factors lead to a decline in the ability of bone formation, accompanied by an increase...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/113A61K45/00A61P19/10
CPCA61K45/00A61P19/10C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 杨承刚向常娟
Owner GUAN BOJIAN BIOTECH CO LTD
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