Compound for delaying senescence of human mesenchymal stem cells and application of same

A technology of compounds and stem cells, applied in the direction of effective ingredients of hydroxyl compounds, medical preparations containing active ingredients, drug combinations, etc., can solve the problems that the effect of quercetin has not been reported yet

Active Publication Date: 2018-12-07
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the role of quercetin in human progeria and mesenchymal stem cells in children with progeria has not been reported yet.

Method used

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  • Compound for delaying senescence of human mesenchymal stem cells and application of same
  • Compound for delaying senescence of human mesenchymal stem cells and application of same
  • Compound for delaying senescence of human mesenchymal stem cells and application of same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1, the screening of natural product library

[0055] This example involves the screening of a natural product library (133 compounds, purchased from Selleck, Cat. No. L1400) on human progeria mesenchymal stem cells (WS MSCs). The method used was the MTS method, and the reagents were purchased from Promega. The quercetin used in subsequent experiments was quercetin dihydrate, which was purchased from Selleck Company.

[0056] The specific method is as follows: Firstly, the fifth-generation WS MSCs were seeded in a 96-well plate, 3000 cells / well, cultured overnight, and replaced with MSC medium containing 1 μM of various compounds on the second day, cultured for 7 days, and the next day Change fluid. The negative control was the solvent dimethyl sulfoxide (DMSO, 1‰), and the positive control was Vitamin C (VC, 280 μM). After 7 days of drug treatment, MTS detection was carried out, the absorbance value (OD value) at 490 nm was measured, and the cell viability...

Embodiment 2

[0059] Embodiment 2, the effective concentration detection of preferred 10 small molecule compounds

[0060] After screening, 10 small molecules such as quercetin, rhein, luteolin, formononetin, honokiol, berberine, kaempferol, phylloxine, synephrine, licochalcone A, etc. were obtained , and studied the effective concentration at the level of WS MSCs cells. see results image 3 .

[0061] Detection method Cell counting. The fifth generation (p5) WS MSCs were seeded in a 6-well plate at 30,000 cells / well, cultured overnight, and replaced with medium containing 100nM, 250nM, 500nM and 1μM of various compounds on the next day, and cultured From about 20 days to the sixth passage, the medium was changed every other day (fresh MSC medium containing various compounds at different concentrations). The negative control was the solvent dimethyl sulfoxide (DMSO). After drug treatment to the sixth passage, cells were counted and analyzed after trypan blue staining. It was found tha...

Embodiment 3

[0081] Example 3. Drug-treated WS MSCs have better osteogenesis, cartilage differentiation ability and anti-aging phenotype.

[0082] Using quercetin 100nM and negative control DMSO (1‰) to treat the seventh generation of WS MSCs (drug treatment since the fifth generation), differentiate into osteoblasts, cartilage and adipocytes ( Figure 5 ). The results showed that the ability of WS MSCs to differentiate into osteoblasts and chondrocytes was enhanced after quercetin treatment.

[0083] Osteogenic Medium:

[0084]

[0085]

[0086] Cartilage Medium:

[0087]

[0088] Fat Medium:

[0089]

[0090] At the same time, progeria mesenchymal stem cells treated with quercetin had a phenotype of delaying aging ( Image 6 ). A, Ki67 immunofluorescence staining of progeria mesenchymal stem cells treated with quercetin, the rate of Ki67 positive cells increased after quercetin treatment; B, Western blot protein quantification of senescence markers p16 and p21 proteins. ...

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Abstract

The invention discloses the applications of compounds, selected from quercetin, chrysophanic acid, digicitrine, formononetin, honokiol, berberine, kaempferol, bergenin, deoxyepinephrine and licochalcone A, as well as derivatives thereof, in preparation of medicines applied to methods that are used for promoting stem cell proliferation, delaying senescence of stem cells, promoting differentiation of the stem cells into osteoblasts, chondrocytes and adipocytes, enhancing the vascularization capability of the stem cells and / or improving exercise tolerance of mice. The invention also discloses that the compounds can be combined with vitamin C for application.

Description

technical field [0001] The present invention relates to the field of biotechnology, more specifically, relates to a series of compounds that delay the aging of human mesenchymal stem cells, promote the proliferation of stem cells, promote the differentiation of stem cells into osteogenesis and chondrocytes, promote the regeneration of blood vessels of mesenchymal stem cells, and enhance the exercise endurance of mice. its use. Background technique [0002] Population aging is an increasingly serious social problem facing the world, especially China, which brings a heavy burden to society and families and affects the sustainable development of society. Therefore, how to delay aging and achieve "healthy aging" is a necessary condition to ensure long-term social stability and sustainable economic development. Therefore, aging and aging-related diseases have become hotspots and difficulties that urgently need to be solved and breakthroughs in the global scientific research fiel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/352A61K31/122A61K31/05A61K31/4375A61K31/366A61K31/12A61K31/137A61P39/06A61P43/00
CPCA61P39/06A61P43/00A61K31/05A61K31/12A61K31/122A61K31/137A61K31/352A61K31/366A61K31/4375
Inventor 曲静刘光慧耿令令张维绮宋默识任若通刘尊鹏
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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