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Applications of non-coding RNA target and inhibitor thereof in preparing medicines capable of promoting osteogenic differentiation of mesenchymal stem cells

A bone marrow mesenchymal and osteogenic differentiation technology, applied in bone/connective tissue cells, animal cells, vertebrate cells, etc.

Active Publication Date: 2017-12-22
深圳盛皓生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The applicant searched and found no ginsenoside RK in the prior art 1 , RG 5 and RZ 1 Related reports of its gene target miR-498 and osteogenic differentiation of bone marrow mesenchymal stem cells

Method used

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  • Applications of non-coding RNA target and inhibitor thereof in preparing medicines capable of promoting osteogenic differentiation of mesenchymal stem cells
  • Applications of non-coding RNA target and inhibitor thereof in preparing medicines capable of promoting osteogenic differentiation of mesenchymal stem cells
  • Applications of non-coding RNA target and inhibitor thereof in preparing medicines capable of promoting osteogenic differentiation of mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Relationship between osteogenic differentiation of bone marrow mesenchymal stem cells and expression level of non-coding RNA miR-498

[0028] 1. Test material

[0029] α-MEM cell culture medium, Hyclone company;

[0030] Fetal bovine serum (FBS), Gibco company;

[0031] Low-sugar DMEM medium, Gibco company;

[0032] miRcute miRNA extraction and isolation kit, Tiangen Biochemical Technology (Beijing) Co., Ltd.;

[0033] TaKaRa Reverse Transcription Kit, TaKaRa Company;

[0034] TaKaRa RT-PCR kit, TaKaRa company;

[0035] BCA kit, Biyuntian Bio;

[0036] Runx2, OCN, β-actin primary and secondary antibodies, Santa Cruz, USA;

[0037] PCR primers were designed and synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd.

[0038] 2. Test method

[0039] 2.1 Isolation and culture of human bone marrow mesenchymal stem cells (hBMSCs)

[0040] According to the standard bone marrow puncture procedure, 5-10 mL of bone marrow was extracted from the post...

Embodiment 2

[0081] Example 2: Ginsenoside RK 1 , RG 5 and RZ 1 Downregulation of miR-498 expression promotes osteogenesis of bone marrow mesenchymal stem cells change

[0082] 1. Test material

[0083] α-MEM cell culture medium, Hyclone company;

[0084] Fetal bovine serum (FBS), Gibco company;

[0085] Low-sugar DMEM medium, Gibco company;

[0086] miRcute miRNA extraction and isolation kit, Tiangen Biochemical Technology (Beijing) Co., Ltd.;

[0087] TaKaRa Reverse Transcription Kit, TaKaRa Company;

[0088] TaKaRa RT-PCR kit, TaKaRa company;

[0089] BCA kit, Biyuntian Bio;

[0090] Runx2, OCN, β-actin primary and secondary antibodies, Santa Cruz, USA;

[0091] Entrust Shanghai Gemma Pharmaceutical Technology Co., Ltd. to design and synthesize PCR primers;

[0092] Ginsenoside RK 1 , RG 5 and RZ 1 Self-made, the purity is greater than 98%.

[0093] 2. Test method

[0094] 2.1 Isolation and culture of human bone marrow mesenchymal stem cells (hBMSCs)

[0095] With ...

Embodiment 3

[0136] Example 3: Overexpression experiments verify the correlation between osteogenic differentiation of bone marrow mesenchymal stem cells and miR-498 targets

[0137] 1. Test material

[0138] α-MEM cell culture medium, Hyclone company;

[0139] Fetal bovine serum (FBS), Gibco company;

[0140] Low-sugar DMEM medium, Gibco company;

[0141] miRcute miRNA extraction and isolation kit, Tiangen Biochemical Technology (Beijing) Co., Ltd.;

[0142] TaKaRa Reverse Transcription Kit, TaKaRa Company;

[0143] TaKaRa RT-PCR kit, TaKaRa company;

[0144] BCA kit, Biyuntian Bio;

[0145] Entrust Shanghai Gemma Pharmaceutical Technology Co., Ltd. to design and synthesize PCR primers, entrust Shanghai Gemma Pharmaceutical Technology Co., Ltd. to design and synthesize miR-498 mimics, miR-498inhibitor, and miR-498 nagative control;

[0146] Ginsenoside RK 1 , RG 5 and RZ 1 Self-made, the purity is greater than 98%.

[0147] 2. Test method

[0148] 2. Test method

[0149] 2.1...

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PUM

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Abstract

The invention relates to applications of a non-coding RNA target and an inhibitor of the non-coding RNA target in preparing medicines capable of promoting osteogenic differentiation of mesenchymal stem cells. According to the technical scheme, for the first time, the result shows that the expression level of miR-498 is relevant to osteogenic differentiation of hBMSCs, and the inhibition for the expression level of miR-498 is beneficial for osteogenic differentiation of hBMSCs. Ginsenosides RK1, RG5 and RZ1 can inhibit the expression level of miR-498 and belong to effective inhibitors for expression of miR-498, ginsenosides RK1, RG5 and RZ1 promote the osteogenic differentiation of hBMSCs by inhibiting the expression of miR-498, and ginsenoside RZ1 has the excellent efficacies.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to the application of a non-coding RNA target and an inhibitor thereof in the preparation of a drug for promoting osteogenic differentiation of bone marrow mesenchymal stem cells. Background technique [0002] The research on the application of bone tissue engineering in the repair of maxillofacial bone tissue defects continues to deepen, and how to construct ideal tissue engineered bone grafts has become a research hotspot. Therefore, human bone marrow mesenchymal stem cells (hMSCs), which are the most widely used in bone tissue engineering, have become a key research object, and studies on osteogenic differentiation of bone marrow mesenchymal stem cells have emerged in an endless stream. [0003] The applicant found that three ginsenosides can promote osteogenic differentiation of bone marrow mesenchymal stem cells, namely ginsenoside RK 1 , RG 5 and RZ 1 . Ginsenoside RK 1 ...

Claims

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Application Information

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IPC IPC(8): C12N5/077
CPCC12N5/0654C12N2500/30
Inventor 孙玉鹤杨勇彭雨泽
Owner 深圳盛皓生物科技有限公司
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