Application of miR-489-3p to preparation of medicines for diagnosing and treating human osteoporosis

A technology for osteoporosis and osteoporosis, applied in the fields of molecular biology and medicine, can solve problems not mentioned

Inactive Publication Date: 2016-10-12
NORTHWESTERN POLYTECHNICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The literature is limited to the study of one target molecule of...

Method used

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  • Application of miR-489-3p to preparation of medicines for diagnosing and treating human osteoporosis
  • Application of miR-489-3p to preparation of medicines for diagnosing and treating human osteoporosis
  • Application of miR-489-3p to preparation of medicines for diagnosing and treating human osteoporosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Detection of low expression of miR-489-3p in bone tissue samples of senile osteoporosis patients by using human miRNA expression profiling chip and qPCR technology.

[0028] (1) Design and prepare specific primer hsa-miR-489-3p, the sequence is shown in Table 1;

[0029] (2) Collection of bone tissue samples from senile osteoporosis patients:

[0030] A total of 111 bone tissue samples from senile osteoporosis patients were collected, immediately preserved in RNA protective agent, and frozen in a -80°C refrigerator. Combined with the patient's past medical history, treatment history and other information, the bone tissue samples of patients without other serious chronic diseases were screened and divided into 90-year-old group and 65-year-old group.

[0031] (3) Extract RNA from bone tissue samples of patients with senile osteoporosis:

[0032] Take the bone tissue sample, peel off the muscle, grind the bone tissue with liquid nitrogen, transfer it to a 1.5ml...

Embodiment 2

[0039] Example 2: Using qPCR technology to detect the low expression of miR-489-3p in the bone tissue of an aged osteoporosis mouse model.

[0040] (1) Design and prepare specific primer mmu-miR-489-3p, the sequence is shown in Table 2;

[0041] (2) Establish an aging mouse model of osteoporosis:

[0042] C57BL / 6 experimental mice were selected and raised to 15 months, 17 months, and 21 months. The mice were sacrificed, tibia samples were collected, and stored in a -80°C refrigerator.

[0043] (3) RNA was extracted from the mouse tibia sample, and the expression level of miR-489-3p in the bone tissue of the mouse model was detected by qPCR technology (the steps were the same as above). The sequence of the mmu-miR-489-3p primer used is as follows:

[0044] Table 2 mouse miR-489-3p primer sequence

[0045]

[0046] refer to figure 2 , the results showed that miR-489-3p was low expressed in the bone tissue of aged osteoporosis mouse model, further indicating that miR-489-...

Embodiment 3

[0047] Example 3: Using qPCR technology to detect the low expression of miR-489-3p in the bone tissue of the HLU hindlimb unloaded osteoporosis mouse model.

[0048] (1) Establish HLU hindlimb unloading osteoporosis mouse model:

[0049] The C57BL / 6 experimental mice were selected, and the mice were hung on the rings of the tail suspension device with a paper clip, and the mouse spine was 30° from the ground to carry out tail suspension and hindlimb unloading. During the experiment, the animals had free access to food and water, and the mice were sacrificed 28 days later.

[0050] (2) RNA was extracted from the mouse tibia sample, and the expression level of miR-489-3p in the bone tissue of the HLU hindlimb unloaded osteoporosis mouse model was detected by qPCR technology (the steps were the same as above).

[0051] refer to image 3 , the results showed that miR-489-3p was lowly expressed in the bone tissue of the HLU hindlimb unloaded osteoporosis mouse model, which fully ...

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Abstract

The invention discloses application of miR-489-3p to preparation of medicines for diagnosing and treating human osteoporosis. According to the technical scheme, the detection of thighbone tissue samples of elderly patients with osteoporosis shows that miR-489-3p is low in expression in human osteoporosis tissue samples and can be used as a marker gene for diagnosing osteoporosis; an analogue and an antagonist of miR-489-3p are designed and prepared, then the analogue and the antagonist of the miR-489-3p are transfected into mice pre-osteoblasts MC3T3-E1 for carrying out a mineralization-alizarin red s dyeing experiment and an ALP dyeing experiment; technical detection of qPCR and Western blot shows that miR-489-3p can accelerate osteocyte differentiation and also can carry out targeted regulation on non-receptor protein-tyrosine-phosphatase6 (PTPN6) molecules. The application of miR-489-3p to preparation of medicines for diagnosing and treating human osteoporosis shows that miR-489-3p can regulate PTPN6 molecules to be used as target sites of action of medicines for treating skeleton system diseases such as osteoporosis.

Description

technical field [0001] The invention belongs to the fields of molecular biology and medicine, and particularly relates to an application of miR-489-3p in preparing medicines for diagnosing and treating human osteoporosis. Background technique [0002] MicroRNAs (miRNAs) are a class of endogenous non-coding single-stranded RNA molecules with a length of about 22-25 nucleotides, which can bind to the 3' non-coding region (3'UTR) of target genes through complementary base pairing. Thereby inhibiting translation or degrading target genes. MicroRNAs are highly conserved during evolution, widely distributed in organisms, and participate in many complex biological processes, such as embryonic development, cell proliferation, apoptosis, fat metabolism, bone formation, etc. However, the biological functions of only a small number of miRNAs have been elucidated. Osteoblasts are the main functional cells for bone formation. As people age, bone development also undergoes dynamic chan...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11A61K48/00A61K31/7105A61P19/10A61P19/08A61P19/02
CPCA61K31/7105C12N15/1137C12N2310/141C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 骞爱荣胡丽芳印崇张琰马剑华赵帆陈志浩李迪杰李思宇马小莉仇伍霞杨超飞王牌苏佩红
Owner NORTHWESTERN POLYTECHNICAL UNIV
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