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Application of protein acetylation enzyme SIRT1 (Silent Mating type Information Regulation 2Homolog1) in promoting proliferation and delaying senility of mesenchymal stem cells

A deacetylase and stem cell technology, which is applied in the application of protein deacetylase SIRT1 in promoting the proliferation of mesenchymal stem cells and delaying aging, and can solve problems such as cell aging and MSC proliferation stagnation.

Active Publication Date: 2013-09-18
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, during the in vitro expansion and culture process, the proliferation of MSCs quickly stagnates (growth arrest), and the cells gradually age, which is currently the main obstacle affecting the application of MSCs.

Method used

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  • Application of protein acetylation enzyme SIRT1 (Silent Mating type Information Regulation 2Homolog1) in promoting proliferation and delaying senility of mesenchymal stem cells
  • Application of protein acetylation enzyme SIRT1 (Silent Mating type Information Regulation 2Homolog1) in promoting proliferation and delaying senility of mesenchymal stem cells
  • Application of protein acetylation enzyme SIRT1 (Silent Mating type Information Regulation 2Homolog1) in promoting proliferation and delaying senility of mesenchymal stem cells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Regulation of SIRT1 on proliferation and aging of mesenchymal stem cells (MSCs)

[0032] 1. Detection of the relationship between the expression level of SIRT1 and the proliferation and senescence of MSCs

[0033] MSCs were isolated and amplified from human bone marrow and adipose tissue by conventional density gradient centrifugation combined with adherent culture, and continued to be subcultured in vitro until aging. Different donors (donors 1, 2, 3) and different passages (4, 3) were collected. 7, 8, 12, 14, and 17 generations) cells, the expression of SIRT1 was detected (the expression level was detected by Western Blot method, the primary antibody was rabbit anti-human SIRT1 antibody (purchased from Epitomics), and the secondary antibody was goat anti-rabbit antibody ( purchased from Zhongshan Jinqiao Company) and MSC proliferation (cell count, PI staining and Brdu staining) and senescence (SA-β-Gal staining) indicators to analyze whether there is a corr...

Embodiment 2

[0104] Example 2. Regulatory mechanism of SIRT1 on mesenchymal stem cell (MSC) proliferation and aging

[0105] Figure 6 The mechanism of cell senescence known so far is reviewed: pathway 1: telomere shortening, oncogene activation and reactive oxygen species cause DNA damage, damage signal activates p53, p53 activates p21Cip1, p21 Cip1 inhibits CDK2 and leads to pRb dephosphorylation; Certain senescence signals in one pathway activate p15INK4b or p16INK4a, and p15INK4b or p16INK4a inhibit CDK4 / 6 leading to dephosphorylation of pRb. It is already known that dephosphorylated pRb and acetylated pRb bind to the transcription factor E2F on the one hand to inhibit the transcription of proliferation-promoting genes, and on the other hand affect the chromatin structure of proliferation-promoting genes to shut down their expression, while hyperphosphorylated pRb and deacetylated pRb The ability of acetylated pRb to bind to E2F is reduced, allowing E2F to be released to promote the t...

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Abstract

The invention discloses an application of protein acetylation enzyme SIRT1 (Silent Mating type Information Regulation 2Homolog1), namely promoting proliferation and delaying senility of mesenchymal stem cells. The experimental result shows that the SIRT1 is needed by the in vitro proliferation of the mescenchymal stem cells, and the condition is verified by MSC (mesenchymal stem cells from marrow) and MSC from fat, and the proliferation and the senility of the MSC can be regulated by regulating the expression level of SIRT1 in the MSC. The invention has very important significances on the determination of the regulation mechanism of the proliferation and the senility of the MSC, the prolonging of the service life of the MSC, and the histocyte substitution and repairing treatment based on the MSC, and is broad in application prospect.

Description

technical field [0001] The invention belongs to the new application of enzymes, in particular to the application of protein deacetylase SIRT1 in regulating the proliferation and aging of mesenchymal stem cells. Background technique [0002] SIRT1 (silent mating type information regulation 2homolog 1), a member of the Sirtuin family (SIRT1-7), is a highly conserved multifunctional NAD-dependent protein deacetylase from bacteria to humans. It prolongs the lifespan of lower organisms such as yeast, fruit flies and nematodes. In higher animals such as mice and humans, it participates in the regulation of various life activities such as malignant tumor formation and chemotherapy resistance, stress response, cell metabolism and aging, so as to treat malignant tumors, diabetes, vascular aging and Parkinson's diseases potential target has attracted widespread attention. [0003] Bone marrow or adipose-derived mesenchymal stem cells (MSCs) have the following advantages: 1) It is co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775C12N5/10C12N15/867
Inventor 裴雪涛岳文袁红丰翟超
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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