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Application of rehmanthin in promoting the proliferation of cik cells in vitro

A technology of rehmanthin and cell body, applied in the field of medicine, can solve problems that have not yet been reported in research

Inactive Publication Date: 2019-02-15
广东梵胜生命科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are no research reports on the proliferation of CIK cells in vitro with the above four rehmanthins and their extracts as main components.

Method used

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  • Application of rehmanthin in promoting the proliferation of cik cells in vitro
  • Application of rehmanthin in promoting the proliferation of cik cells in vitro
  • Application of rehmanthin in promoting the proliferation of cik cells in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Effects of Rehmanthin B, Rehmanthin D, Pyrothin A and Pyrothin B on the Proliferation of CIK Cells in Vitro

[0024] 1. Experimental materials

[0025] Rehmanthin B, rehmanthin D, keratin A and keratin B are self-made, and the purity is above 95%.

[0026] 2. Experimental method

[0027] 1. Isolation of single cells

[0028] Use Ficoll-Hypaque density gradient centrifugation to separate PBMC from the peripheral blood of healthy volunteers, add 4mL PBS to dilute 4mL whole blood (ratio 1:1), take out the diluted blood and slowly add it to a 4mL Ficoll-Hypaque centrifuge tube, be careful not to Disturb the liquid surface interface, centrifuge at 2000r / min for 20min, and gently draw the mononuclear cells into another centrifuge tube. Add 5 mL of normal saline, blow the cells evenly, and centrifuge to wash twice.

[0029] 2. Culture of CIK cells

[0030] Add CIK initial culture medium to the PBMC suspended in normal saline, and adjust the cell concentration t...

Embodiment 2

[0040] Example 2: Preparation and activity of Rehmannia glutinosa extract with rehmanthin B, rehmanthin D, and pyroxanthin A as main components

[0041] The preparation method comprises the following steps:

[0042] Step S1, wash and slice fresh rehmannia glutinosa, dry at 55°C to obtain fresh rehmannia glutinosa slices; extract fresh rehmannia glutinosa slices with 95% ethanol aqueous solution under heat reflux for 3 times, each time for 2 hours, with a solid-to-liquid ratio of 1:20, combine the extracts, Concentrate under reduced pressure to obtain extract;

[0043] Step S2, dissolving the extract with an appropriate amount of water, first extracting with equal volume of petroleum ether for 3 times to remove impurities, then extracting with dichloromethane for 3 times, combining the dichloromethane extracts, concentrating and drying to obtain the dichloromethane extract;

[0044] Step S3, dissolve the dichloromethane extract with 30% ethanol aqueous solution, mix the resin ...

Embodiment 3

[0058] Example 3: Separation and preparation of rehmanthin B, rehmanthin D, and pyrothin A monomers

[0059]The separation preparation method comprises the steps:

[0060] Step S1, wash and slice fresh rehmannia glutinosa, dry at 55°C to obtain fresh rehmannia glutinosa slices; extract fresh rehmannia glutinosa slices with 95% ethanol aqueous solution under heat reflux for 3 times, each time for 2 hours, with a solid-to-liquid ratio of 1:20, combine the extracts, Concentrate under reduced pressure to obtain extract;

[0061] Step S2, dissolving the extract with an appropriate amount of water, first extracting with equal volume of petroleum ether for 3 times to remove impurities, then extracting with dichloromethane for 3 times, combining the dichloromethane extracts, concentrating and drying to obtain the dichloromethane extract;

[0062] Step S3, dissolve the dichloromethane extract with 30% ethanol aqueous solution, mix the resin and load it on the XDA-1B macroporous adsorp...

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Abstract

The invention discloses application of rehmannin in promoting in-vitro proliferation of CIK (cytokine-induced killer) cells. The application has the advantages that rehmannin B, rehmannin D and jioglutin A can obviously improve the in-vitro proliferation function of the CIK cells, but jioglutin B does not have the in-vitro proliferation function; compared with the jioglutin A, only the configuration of Cl-connected carbon atoms in jioglutin B is different, and the configurations of carbon atoms in the rehmannin B and the rehmannin D are the same with the configuration of carbon atoms in the jioglutin A, so that the configuration of carbon atoms is critical to the promotion of cell proliferation activity of the CIK cells of the rehmannin type compounds; accordingly, the rehmannin B, the rehmannin D and the jioglutin A can be used for preparing a culture medium for promoting the in-vitro proliferation of the CIK cells, for example, the culture medium can be prepared by adding the rehmannin B, the rehmannin D or the jioglutin A into a RPMI1640 culture solution.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to the application of rehmanthin in promoting the proliferation of CIK cells in vitro. Background technique [0002] Rehmanthin B, Rehmanthin D, Pyrothaxanthin A and Pyrorethin B are natural products isolated from Rehmannia glutinosa. The chemical structures are very similar. The chemical information is shown in the table below. [0003] [0004] At present, there are no research reports on the proliferation of CIK cells in vitro with the above four rehmanthins and their extracts as main components. Contents of the invention [0005] The purpose of the present invention is to provide the application of rehmanthin in promoting the proliferation of CIK cells in vitro. [0006] Above-mentioned object realizes through following technical scheme: [0007] The application of rehmanthin with the following chemical structure in promoting the proliferation of CIK cells in vitro; ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0638C12N2501/999
Inventor 马海周
Owner 广东梵胜生命科技有限公司
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