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Application of AMPK activator in preparation of product for improving abnormal AMPK signaling pathway in bone marrow mesenchymal stem cells

A bone marrow mesenchymal and signaling pathway technology, applied in the biological field, can solve problems such as unclear molecular mechanism and bone mineralization, achieve the effect of restoring osteogenic differentiation ability, promoting differentiation ability, and remarkable effect

Pending Publication Date: 2020-12-25
XIAN TISSUE ENG & REGENERATIVE MEDICINE RES INST
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Problems solved by technology

Clinically, ALPL gene mutations lead to the occurrence of hypoalkaline phosphatasia (HPP), and the clinical phenotype of patients is mainly accompanied by abnormal bone development and severe bone mineralization deficiency.
Since the molecular mechanism of potential pathogenicity of HPP is still not very clear, the current conventional regimens used in the clinical treatment of HPP patients can only slow down or delay the decline of bone mass, and cannot fundamentally solve the primary problem of bone development defects.

Method used

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  • Application of AMPK activator in preparation of product for improving abnormal AMPK signaling pathway in bone marrow mesenchymal stem cells
  • Application of AMPK activator in preparation of product for improving abnormal AMPK signaling pathway in bone marrow mesenchymal stem cells
  • Application of AMPK activator in preparation of product for improving abnormal AMPK signaling pathway in bone marrow mesenchymal stem cells

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Embodiment 1

[0067] 1. Detection of osteogenic differentiation ability of bone marrow mesenchymal stem cells with abnormal osteogenic differentiation ability derived from patients with hypoalkaline phosphatase disease 1. Bone marrow mesenchymal stem cells with abnormal osteogenic differentiation ability derived from patients with hypoalkaline phosphatase disease stem cell acquisition

[0068] Bone marrow mesenchymal stem cells with abnormal osteogenic differentiation ability isolated from hypoalkaline phosphatase patients were adhered to culture medium in α-MEM medium containing 10% fetal bovine serum, and each passage time was 2- After 3 days, 1-3 passages of bone marrow mesenchymal stem cells with abnormal differentiation ability were obtained.

[0069] 2. Detection of osteogenic differentiation ability of bone marrow mesenchymal stem cells with abnormal osteogenic differentiation ability derived from patients with hypoalkaline phosphatase disease

[0070] 1) Detection of osteogenic dif...

Embodiment 2

[0077] 1. Detection of osteogenic differentiation ability of bone marrow mesenchymal stem cells derived from ALPL knockout mice with abnormal osteogenic differentiation ability

[0078] 1. Obtaining bone marrow mesenchymal stem cells with abnormal osteogenic differentiation ability derived from ALPL knockout mice

[0079] Bone marrow mesenchymal stem cells with abnormal osteogenic differentiation ability derived from isolated ALPL knockout mice were adhered to the wall in α-MEM medium containing 20% ​​fetal bovine serum, and the abnormal differentiation ability of passage 0-3 was obtained bone marrow mesenchymal stem cells.

[0080] 2. Detection of osteogenic differentiation ability of bone marrow mesenchymal stem cells derived from ALPL knockout mice with abnormal osteogenic differentiation ability Digested mesenchymal stem cells of the p0 generation from normal and ALPL knockout mice and counted cells at 1× 10 5 and 5×10 5 Inoculate into 24-well plates and 6-well plates f...

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Abstract

The invention provides an application of an AMPK activator in the preparation of a product for improving an abnormal AMPK signaling pathway in bone marrow mesenchymal stem cells. In-vitro experimentsprove that the osteogenic differentiation capacity of bone marrow mesenchymal stem cells derived from patients with hypophosphatasia or ALPL knockout mice is abnormal, a signal channel protein chip finds that the intracellular AMPK protein phosphorylation of the bone marrow mesenchymal stem cells with the abnormal osteogenic differentiation capacity is remarkably reduced, and the AMPK activator metformin is used to act on the bone marrow mesenchymal stem cells with the abnormal osteogenic differentiation capacity, so that a blocked downstream pathway is restarted, and thereby the osteogenic differentiation characteristic of the bone marrow mesenchymal stem cells with the abnormal osteogenic differentiation capacity is recovered.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of an AMPK activator in the preparation of a product for improving the abnormality of the AMPK signaling pathway in bone marrow mesenchymal stem cells. Background technique [0002] Tissue-nonspecific alkaline phosphatase (tissue-nonspecific alkaline phosphatase, TNAP) is a tissue-nonspecific alkali encoded by the liver / bone / kidney alkaline phosphatase gene (liver / bone / kidney alkaline phosphatase, ALPL) Phosphatase, which mainly releases monophosphate by hydrolyzing the phosphate bond of pyrophosphate or polyphosphate to dephosphorylate. Studies have confirmed that tissue non-specific alkaline phosphatase utilizes phosphatase properties in vivo to achieve detoxification by removing LPS phosphate groups. In addition, monophosphate combined with calcium ions released from the hydrolysis of pyrophosphate by tissue non-specific alkaline phosphatase deposits in the bone e...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K31/155A61P19/08A61P19/10
CPCC12N5/0663A61K31/155A61P19/08A61P19/10C12N2501/999
Inventor 刘文佳张立强
Owner XIAN TISSUE ENG & REGENERATIVE MEDICINE RES INST
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