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Application of oligonucleotide compound of micro-RNA (ribonucleic acid) associated with senile osteoporosis

An oligonucleotide and osteoporosis technology, applied in the application field of oligonucleotide compounds, can solve the lack of animal experiment data support, lack of microRNA oligonucleotides, and lack of microRNA oligonucleotides for senile osteoporosis Nucleotide and other issues to achieve the effect of promoting osteoblast function and inhibiting degree

Inactive Publication Date: 2016-11-30
NORTHWESTERN POLYTECHNICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this invention only uses myoblasts for osteogenesis induction, and does not use bone-derived cells such as MC3T3-E1 pre-osteoblasts as research objects, and also lacks data support from animal experiments
[0006] In summary, based on the existing technologies and methods, on the one hand, there is a lack of microRNA oligonucleotides that can promote osteoblast function and bone formation; on the other hand, in the process of practical application, more osteoporotic Osteoporosis occurs in old age, and there is currently a lack of microRNA oligonucleotides that can inhibit senile osteoporosis

Method used

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  • Application of oligonucleotide compound of micro-RNA (ribonucleic acid) associated with senile osteoporosis
  • Application of oligonucleotide compound of micro-RNA (ribonucleic acid) associated with senile osteoporosis
  • Application of oligonucleotide compound of micro-RNA (ribonucleic acid) associated with senile osteoporosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Expression of microRNAs related to senile osteoporosis in bone tissue of elderly patients.

[0029] (1) Extraction of total RNA from bone tissue.

[0030] Bone tissues discarded during clinical operations were collected and divided into 60-year-old and 90-year-old age groups, as well as ambulatory and elderly bedridden >24-month groups. Immediately after collection, the tissue was frozen and stored in a -80°C refrigerator. When extracting RNA, the bone tissue frozen at -80°C was taken out, placed in a mortar pre-cooled with liquid nitrogen, and quickly ground into powder after adding an appropriate amount of liquid nitrogen for quick freezing. Transfer the powder to Reagents, chloroform extraction, isopropanol precipitation, after washing with 75% ethanol and evaporating to dryness, add RNase free sterile water to dissolve RNA, and use a UV spectrophotometer to perform quality inspection on RNA samples to ensure that A260 / 280≥1.80 of all samples, The tota...

Embodiment 2

[0038] Example 2: miR-138-5p agonists promote MC3T3-E1 preosteoblast function.

[0039] (1) Chemical synthesis of miR-138-5p agonist Agomir-138-5p.

[0040] Obtain the nucleotide sequence of miR-138-5p mature body from miRBase, synthesize oligonucleotide single strand and complementary double strand identical to the mature body sequence, purify by high performance liquid chromatography, and carry out full-strand methylation , before and after 2'-4' site phosphorothioation backbone modification, and 3' end cholesterol labeling, and finally obtained the miR-138-5p double strand with the sequence 5'-agcugguguugugaaucaggccg-3', 5'-gccugauucacaacaccagcuuu-3' Oligonucleotide agonists. At the same time, a random negative control was synthesized with the sequence 5'-guccgaaggugcgcguagaguuu-3', 5'-acucuacgcgcaccuucggacuu-3'. The synthesized oligonucleotide compound was formulated into a 20 μM storage solution with RNase free water and stored at -80°C.

[0041] (2) Agomir-138-5p tran...

Embodiment 3

[0051] Example 3: miR-138-5p inhibitor improves the degree of senile osteoporosis.

[0052] (1) Chemical synthesis of miR-138-5p inhibitor Antagomir-138-5p.

[0053] The nucleotide sequence of the mature body of miR-138-5p was obtained from miRBase, and a single-stranded oligonucleotide complementary to the sequence of the mature body was synthesized, purified by high-performance liquid chromatography, and the whole chain was 2'methylated, 5' Modification of two phosphorothioate sites, modification of four phosphorothioate sites at 3', and cholesterol labeling at the 3' end, finally obtaining miR-138-5p single-stranded oligonucleotide with the sequence 5'-cggccugauucacaacaccagcu-3' Inhibitors. At the same time, a random negative control was synthesized with the sequence 5'-uuguacuacacaaaaguacug-3'. The synthesized oligonucleotide compound was formulated into a 20 μM storage solution with RNase free water and stored at -80°C.

[0054] (2) Antagomir-138-5p inhibits senile ost...

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Abstract

The invention discloses an application of an oligonucleotide compound of micro-RNA (ribonucleic acid) associated with senile osteoporosis. The oligonucleotide compound is used for filling in the gap of micro-RNA based oligonucleotide in application of preparing the compound for promoting osteoblast functions and inhibiting the senile osteoporosis. The technical scheme includes that the micro-RNA associated with the senile osteoporosis is obtained by screening, the oligonucleotide based on the micro-RNA associated with the senile osteoporosis is synthesized and transfected to into preosteoblast and senile osteoporosis mice bodies, results indicate that the osteoblast functions can be promoted, and the progress of the senile osteoporosis is inhibited. The prepared oligonucleotide based on the micro-RNA associated with the senile osteoporosis can inhibit the degree of the senile osteoporosis by promoting the osteoblast functions, and the gap of the micro-RNA based oligonucleotide in application of preparing the compound for promoting the osteoblast functions and inhibiting the senile osteoporosis is filled in.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, in particular to the application of an oligonucleotide compound of microRNA related to senile osteoporosis. Background technique [0002] Senile osteoporosis refers to a disease caused by the decline in the function of the skeletal system with age. It often manifests as a decrease in bone density and bone mass, destruction of bone microstructure, and increased bone fragility and susceptibility to fractures. my country is currently in the process of accelerating population aging. The 2000 census showed that there were about 260 million people over the age of 50, accounting for 20.68% of the total population; according to the 2010 census, the total number of people over the age of 50 rose to about 340 million. It has increased by 30.8% in 10 years (Census Office of the State Council. China 2010 Census Data [M]. Beijing: China Statistics Press). [0003] MicroRNAs are a class of endogenous non-...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 骞爱荣马剑华赵帆陈志浩胡丽芳李迪杰印崇马小莉苏佩红
Owner NORTHWESTERN POLYTECHNICAL UNIV
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