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A ccr5 expression promoter and its application in the preparation of drugs that promote NK cells to penetrate the blood-brain barrier
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A technology of expressing promoters and NK cells, which is applied in the field of medicine and can solve problems such as the absence of NK cells penetrating the blood-brain barrier
Active Publication Date: 2018-08-14
徐州立顺康达医药科技有限公司
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[0007] Ginsenoside RZ has not been seen so far 1 There are no reports on the effect of CCR5 expression on NK cells, let alone its use to promote NK cells to penetrate the blood-brain barrier
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Embodiment 1
[0022] Embodiment 1: Ginsenoside RZ 1 Promote high expression of CCR5 in NK cells
[0023] 1. Experimental method
[0024] 1. Culture and grouping of NK cells
[0025] Take peripheral anticoagulant blood from healthy blood donors, separate mononuclear cells, and use NK medium containing 500 U / ml rhIL-2 (purchased from CellGenix, Germany) to adjust the number of cells to 3.5×10 5 / ml, inoculated in 6-well culture plate, 5ml per well, at 37°C, 5% CO 2 Culture in an incubator, change the medium once every 2 days, and adjust the number of cells to 5×10 4 / ml, collect NK cells cultured for 10 days; randomly divide them into the following four groups, and use different media to continue treatment for 24 hours:
[0026] Ginsenoside RZ 1 Group: NK medium supplemented with 20 μM ginsenoside RZ 1 ;
[0027] Ginsenoside RK 1 Group: NK medium supplemented with 20 μM ginsenoside RK 1 ;
[0028] Ginsenoside RG 5 Group: NK medium supplemented with 20 μM ginsenoside RG 5 ;
[0...
Embodiment 2
[0043] Embodiment 2: Ginsenoside RZ 1 Promote NK cell penetration into human brain microvascular endothelial cells (HBMEC) model
[0044] 1. Experimental method
[0045] 1. Culture and grouping of NK cells
[0046] With embodiment 1.
[0047] 2. Culture of HBMECs
[0048] HBMEC is provided by the Cell Biology Laboratory of China Medical University, and can be cultured according to conventional methods, specifically: add HBMEC to RPMI-1640 nutrient solution containing 10% calf serum, 10% Nuserum, 2mmol / L glutamine, 1mmol / L sodium pyruvate, non-essential amino acids, MEM vitamins, at 37°C, 5% CO 2, cultured in a cell culture incubator with 95% humidity. Replace the fresh culture medium every 2 days. After the cells grow to about 90% on the bottom of the culture flask, digest the cells with cell digestion solution containing 0.25% trypsin, then add the culture medium, blow into a single-cell suspension, and passage in 1:4. into a new culture bottle.
[0049] 3. Establis...
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Abstract
The invention relates to a CCR5 expression accelerant and application thereof to preparation of medicines for promoting NK cells to penetrate through a blood brain barrier. The invention discovers that ginsenoside RZ1 can promote the NK cells to highly express CCR5 to promote the NK cells to penetrate through the blood brain barrier, so that the ginsenoside RZ1 can be used for preparation of medicines for promoting the NK cells to express CCR5 and medicines for promoting the NK cells to penetrate through the blood brain barrier.
Description
technical field [0001] The invention relates to the field of medicines, in particular to a CCR5 expression promoter and its application in the preparation of medicines for promoting NK cells to penetrate the blood-brain barrier. Background technique [0002] Natural killer cells (NK cells) are one of the important components of the body's innate immune system. They can kill target cells without specific antigen stimulation, and have the functions of immune clearance and immune surveillance. Only activated NK cells can spread and infiltrate into the source of infection or tumor tissue to play an anti-tumor effect. NK cells can also secrete a variety of cytokines, such as TNF-α, TNF-β, IFN-γ, etc. to cooperate with them in anti-tumor. The activation of NK cells is regulated by their surface receptors. These receptors transmit inhibitory or activating signals according to the structure and function to regulate the activity of NK cells, thereby regulating the anti-tumor functio...
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