Osteoporosis diagnosis marker and nucleic acid medicine for promoting osteoporosis bone regeneration

A diagnostic marker and osteoporosis technology, applied in the field of biomedicine, can solve problems such as osteoporosis and bone metabolism that have not been reported in the field, and achieve the effect of preventing and treating osteoporosis

Active Publication Date: 2021-05-04
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, piRNA, as a new non-coding RNA molecule, has not been reported in the field of osteoporosis and bone metabolism

Method used

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  • Osteoporosis diagnosis marker and nucleic acid medicine for promoting osteoporosis bone regeneration
  • Osteoporosis diagnosis marker and nucleic acid medicine for promoting osteoporosis bone regeneration
  • Osteoporosis diagnosis marker and nucleic acid medicine for promoting osteoporosis bone regeneration

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] In this embodiment, the following related researches are done on BMSCs piRNA of osteoporosis rats:

[0068] piRNA sequencing was performed on BMSCs of osteoporotic rats, and the obtained piRNA expression volcano map is shown in Figure 2(A);

[0069] Then, the differential expression analysis of piRNAs found by piRNA sequencing showed that 28 piRNAs were significantly differentially expressed in BMSCs of osteoporotic rats, including 13 up-regulated differentially expressed piRNAs and 15 down-regulated differentially expressed piRNAs;

[0070] Among them, the heat map of significantly differentially expressed piRNAs is shown in Figure 2 (B);

[0071] The up-regulated differentially expressed piRNAs and the down-regulated differentially expressed piRNAs were verified by qPCR, and the results are shown in Figure 2(C) and Figure 2(D) (wherein, n=10, *p<0.05); thus determined Several piRNAs related to osteoporosis including piRNA-63049 were identified.

Embodiment 2

[0073] This example is used to study the correlation between the expression level of piR-63049 and the severity of osteoporosis. Specific steps are as follows:

[0074] (1) Construct the rat model of ovariectomized osteoporosis and sham operation group, perform microCT scanning at 0w, 4w, 8w, and 12w after operation, and calculate bone mass and bone microstructure-related parameters (including bone density BMD, relative bone volume BV / TV, number of trabecular bone and thickness of trabecular bone, etc.);

[0075] (2) Extract rat BMSCs at the above time points, detect the expression level of piR-63049 by qPCR, and analyze the correlation between the expression level of piR-63049 and the severity of osteoporosis through statistical analysis;

[0076] The specific results are as follows:

[0077] The nuclear and cytoplasmic separation marker identification structure is shown in Figure 3 (A), which shows the expression levels of the internal reference GAPDH and nuclear lamin (L...

Embodiment 3

[0082] This example is used to explore the target of piR-63049. According to the results of functional bioinformatics analysis of piRNA, piR-63049 may inhibit the osteogenic differentiation of BMSCs by inhibiting the expression of Wnt2b in the cytoplasm. Therefore, in this example, a luciferase reporter gene plasmid containing the 3'-UTR end of Wnt2b was constructed to verify whether Wnt2b is the target of piR-63049.

[0083] The specific results are as follows:

[0084] As shown in Figure 3(E), the expression level of Wnt2b gradually increased with time during the osteogenic differentiation of BMSCs; as shown in Figure 3(F), the network diagram of Wnt2b protein interaction, in which the solid line indicates the common pathway The dotted line indicates the binding relationship; it can be seen from the figure that the Wnt2b protein is closely related to the expression of various genes;

[0085] As shown in Figure 3(G), after Wnt2b was knocked down, the expression of β-catenin ...

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Abstract

The invention provides an osteoporosis diagnostic marker and a nucleic acid medicine for promoting osteoporosis bone regeneration. The osteoporosis diagnosis marker is Piwi protein binding RNA-63049, and has a nucleotide sequence as shown in SEQ ID NO. 1. The nucleic acid medicine for promoting osteoporotic bone regeneration is an antagonist of Piwi protein binding RNA-63049. After the piwi protein is combined with piRNA-63049, a piR-63049/piwi protein complex is formed, so that the expression of the Wnt2b protein is inhibited, and further, the osteogenic differentiation capability of the BMSCs is weakened; and the piRNA-63049 antagonist plays roles in promoting bone regeneration and preventing osteoporosis bone mass loss in vivo, so that the piRNA-63049 antagonist can be used as a nucleic acid medicine for promoting osteoporosis bone regeneration.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a medicine for treating osteoporosis and its application, in particular to a diagnostic marker for osteoporosis and a nucleic acid medicine for promoting bone regeneration of osteoporosis. Background technique [0002] With the intensification of the aging population in my country, osteoporosis in the elderly or postmenopausal women has become an important health problem for the whole society. Osteoporosis can lead to complications such as general bone pain, short stature, and vertebral and hip fractures. In addition to seriously affecting the activities and quality of life of the elderly, these complications can also cause severe disability and even death. Osteoporosis has become the most common geriatric disease after hypertension, diabetes, and cardiovascular and cerebrovascular diseases, bringing a huge burden to both the family and the society. The pathophy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113A61K31/7105A61P19/10
CPCC12Q1/6883A61K31/7105A61P19/10C12Q2600/178C12Q2600/158
Inventor 彭松林王尚
Owner SHENZHEN PEOPLES HOSPITAL
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