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Preparation of a secoiridoid glycoside compound and its application in improving the viability of bone marrow mesenchymal stem cells

A technology of split iridoid glycosides and complexes, which is applied in the preparation of split iridoid glycoside complexes and the application field of improving the vitality of bone marrow mesenchymal stem cells, and can solve the problems of cell morphology, metabolism, proliferation and differentiation Potential changes, BMSCs functional damage and other issues, to achieve the effect of maintaining multi-directional differentiation potential, increasing ability, and improving proliferation ability

Active Publication Date: 2021-03-23
SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the influence of various physical and chemical factors in the process of in vitro expansion and culture, BMSCs often lead to changes in cell morphology, metabolism, proliferation and differentiation potential. Important cause of functional impairment

Method used

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  • Preparation of a secoiridoid glycoside compound and its application in improving the viability of bone marrow mesenchymal stem cells
  • Preparation of a secoiridoid glycoside compound and its application in improving the viability of bone marrow mesenchymal stem cells
  • Preparation of a secoiridoid glycoside compound and its application in improving the viability of bone marrow mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0030] Embodiment: A split iridoid glycoside compound, the split iridoid glycoside compound is composed of 5~80 μM oleuropein, 10 μM rehmanthin A, 50 μM gentiopicroside and conventional in vitro Prepared as a medium for cell culture. The medium for conventional in vitro cell culture contains necessary cell culture supplements, such as serum, NaHCO 3 Wait.

[0031] The preparation method is as follows: firstly, compound A stock solution containing oleuropein and rehinoside A is prepared; secondly, compound B containing gentiopicroside is prepared with DMEM powder; sterile fetal bovine serum stock solution is used in different ratios (V / V ) Compound A stock solution and compound B are mixed to prepare a split iridoid glycoside compound; the specific steps are:

[0032] (1) Preparation of complex A stock solution: According to Table 1, accurately weigh oleuropein and rehmanthin A in 10mL DMSO, heat and ultrasonically dissolve with an ultrasonic dissolving instrument, the heatin...

experiment example 1

[0039] Experimental Example 1: The effect of the split iridoid glycoside compound of the present invention on the proliferation ability of BMSCs

[0040] Press 0.2×10 4 Cells / well subculture and inoculate BMSCs in 96-well plates, and the next day, divide the adhered BMSCs into control group, reagent 1 group, reagent 2 group, reagent 3 group, reagent 4 group, and reagent 5 group, with at least 3 cells in each group Repeat hole. The reagent group was the secoiridoid glycoside complex prepared according to the preparation schemes described in Table 1 and Table 2, and the control group was replaced with 10% FBS-DMEM, a conventional medium containing the same volume of DMSO as the reagent group. After 48 hours of action, add 10 μL of CCK-8 solution to each well (be careful not to generate air bubbles, so as not to affect the OD value), 37°C 5% CO 2 Incubate in an incubator for 2 to 4 hours, and measure the absorbance at 450 nm with a microplate reader. see results Figure 5 a. ...

experiment example 2

[0042] Experimental Example 2: The detection experiment of the iridoid glycoside compound compound of the present invention on active oxygen in BMSCs

[0043] Press 6×10 4 BMSCs were subcultured into 6-well plates in cells / well, and the attached BMSCs were divided into control group and reagent 3 group the next day. The reagent group 3 is the secoiridoid glycoside compound prepared according to the preparation scheme of the reagent group 3 described in Table 1 and Table 2, and the control group is the conventional medium 10% FBS-DMEM containing the same volume of DMSO as the reagent group . According to this, the medium was changed every 2 days, and the BMSCs were subcultured when the growth reached 80% confluence. After 30 days of action, the active oxygen kit was used for detection. see results Image 6 . The results showed that the active oxygen level of BMSCs in the reagent group 3 was 30% lower than that in the control group, indicating that the split iridoid glycosid...

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Abstract

The invention belongs to the field of biomedicine and provides preparation of a secoiridoid glycoside complex and application of the secoiridoid glycoside complex in improving the viability of bone-marrow mesenchymal stem cells in order to solve the problems about changes in the cellular morphology, metabolism, proliferation and differentiation potential of bone-marrow mesenchymal stem cells (BMSCs) cultured in vitro, improve the proliferation of the BMSCs cultured in vitro and keep the BMSCs good in differentiation potential. The secoiridoid glycoside complex is prepared by mixing oleuropein, rehmaionoside A and gentiopicroside with a conventional in-vitro cell culture medium in a certain proportion. By means of the secoiridoid glycoside complex, the proliferation capacity of the BMSCs can be improved, short-term proliferation is promoted, so that a lot of cells are obtained to be used for transplantation for treatment, and moreover, the survival rate of the transplanted cells in vivo can be improved. When the complex acts on the BMSCs for a long time, the oxidative-damage-resistant ability of the cells can be improved, and the BMSC differentiation tending to neurons is promoted without changing the biological characteristics of the BMSCs. A method is provided for the wide application of the BMSCs in research of medical tissue engineering.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the preparation of a split iridoid glycoside compound and its application in improving the proliferation and differentiation activity of bone marrow mesenchymal stem cells. The split iridoid glycoside compound consists of oleuropein (Oleuropein, OE), rehmaionoside A (Rehmaionoside A) and gentiopicroside (Gentiopicroside) preparation composition. Background technique [0002] There are many kinds of terpenoids in nature, they are widely distributed, and they have a wide range of physiological activities. Among them, iridoids, as monoterpenoids, are acetal derivatives of Iridodial [Sun Yujia, Tang Shuhan, Wang Shanshan, et al. Structural classification and biological Activity [J]. Heilongjiang Science and Technology Information, 2014, (2): 80.]. Iridoids are widely distributed in dicotyledonous plants such as Scrophulariaceae, Rubiaceae, Lamiaceae, Gentianaceae, Verbenaceae...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2500/30
Inventor 陆利刘雪芹牛晓洁杨娜赵雪纯赵云鹤陈显久
Owner SHANXI MEDICAL UNIV
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