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Preparation of secoiridoid glycoside complex and application of secoiridoid glycoside complex in improving viability of bone-marrow mesenchymal stem cells

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

Active Publication Date: 2017-08-25
SHANXI MEDICAL UNIV
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  • Description
  • Claims
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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 secoiridoid glycoside complex and application of secoiridoid glycoside complex in improving viability of bone-marrow mesenchymal stem cells
  • Preparation of secoiridoid glycoside complex and application of secoiridoid glycoside complex in improving viability of bone-marrow mesenchymal stem cells
  • Preparation of secoiridoid glycoside complex and application of secoiridoid glycoside complex in improving viability of bone-marrow mesenchymal stem cells

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Experimental program
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Embodiment

[0030] Example: a schizo iridoid glycoside complex, the schizo iridoid glycoside complex is composed of 5-80 μM oleuropein, 10 μM retinoid A, 50 μM gentiopicroside and conventional in vitro cell culture medium. 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 prepare a stock solution of complex A containing oleuropein and retinoid A; secondly, use DMEM powder to prepare compound B containing gentiopicroside; use sterile fetal bovine serum stock solution, different ratios (V / V ) Compound A stock solution and compound B are mixed to prepare split iridoid glycoside complexes; the specific steps are:

[0032] (1) Preparation of complex A stock solution: According to Table 1, accurately weigh oleuropein and diflavin A in 10 mL of DMSO, and dissolve them in an ultrasonic dissolving instrument, heating temperature is 50 °C, ultrasonic power is 300 W, and...

experiment example 1

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

[0040] Press 0.2×10 4 The cells / well were subcultured and seeded with BMSCs in 96-well plates. The next day, the adherent BMSCs were divided into control group, reagent 1 group, reagent 2 group, reagent 3 group, reagent 4 group, and reagent 5 group, with at least 3 in each group. Duplicate holes. The reagent group was the split iridoid glycoside complexes prepared according to the preparation scheme described in Table 1 and Table 2, and the control group was replaced with a conventional medium containing 10% FBS-DMEM with 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 for 2 to 4 hours in an incubator, and measure the absorbance at 450 nm with a microplate reader. se...

experiment example 2

[0042] Experimental Example 2: Experiment for the detection of reactive oxygen species in BMSCs by the iridoid glycoside compound complex of the present invention

[0043] Press 6×10 4 The cells / well were subcultured and seeded with BMSCs in 6-well plates. The next day, the adherent BMSCs were divided into control group and reagent 3 groups. The reagent group 3 is the split iridoid glycoside complex 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 passaged when the growth reached 80% confluence. After 30 days of action, the reactive oxygen species kit was used for detection. see the results Image 6 . The results showed that the reactive oxygen species level of BMSCs in Reagent 3 group was 30% lower than that in the control grou...

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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 complex and its application for improving the proliferation and differentiation activity of bone marrow mesenchymal stem cells. The split iridoid glycoside complex is composed of oleuropein (Oleuropein, OE), Rehmaionoside A (Rehmaionoside A) and Gentipicroside (Gentiopicroside) are formulated. Background technique [0002] Terpenoids are widely distributed in nature and have a wide range of physiological activities. Among them, iridoids (Iridoids), as monoterpenoids, are the acetal derivatives of Iridodial [Sun Yujia, Tang Shuhan, Wang Shanshan, et al. Structural classification and biology of iridoids Activity [J]. Heilongjiang Science and Technology Information, 2014, (2): 80.]. Iridoids are widely distributed in Scrophulariaceae, Rubiaceae, Lamiaceae, Gentianaceae, Verviaceae, Oleaceae and other dicotyledonous plants. At pres...

Claims

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Application Information

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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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