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Application of ginsenoside Rh2 to inhibition of Treg cells

A technology of ginsenosides and cells, which is applied in the field of application and preparation of ginsenoside Rh2 in inhibiting Treg cells, and can solve problems such as unclear specific mechanism of action

Inactive Publication Date: 2016-02-03
JIANGSU MICROMEDMARK BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ginsenosides can also induce differentiation of tumor cells, but the specific mechanism of action is still unclear

Method used

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  • Application of ginsenoside Rh2 to inhibition of Treg cells
  • Application of ginsenoside Rh2 to inhibition of Treg cells
  • Application of ginsenoside Rh2 to inhibition of Treg cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Embodiment 1: the preparation of ginsenoside Rh2

[0109] 1.1 Preparation of dry powder of protopanaxadiol group saponins

[0110]Take 5-15 mesh ginseng coarse powder, reflux extraction with 40-100% ethanol for 1-4 times, each time for 1-3 hours, collect the extract, filter, and recover ethanol from the filtrate until there is no ethanol smell, then concentrate in vacuum for 1.5-2 hour, the concentrated solution was added with equal volume of distilled water, stirred evenly, filtered, and the filtrate was adsorbed through a macroporous adsorption resin column; water-soluble impurities were washed away with 2-4 times the volume of resin bed volume distilled water, and the resin column for adsorbing ginsenosides was set aside; the above-mentioned adsorption The resin column is eluted with 20-50% ethanol, and the amount of 20-50% ethanol is 1-5 times the volume of the resin bed. Thin-layer chromatography wherein is: developing agent is the chloroform of 30-100: 10-50: 2-2...

Embodiment 2

[0120] Example 2: Rh2 reduces Treg levels in tumor mice

[0121] The mouse model was established by implanting tumors during administration (hereinafter referred to as TIDA); T cells in the whole blood and spleen of mice in each group were extracted; the ratio of Treg to T cells in whole blood and spleen of mice in different groups was detected.

[0122] 2.1. Cell culture

[0123] Under the culture conditions of complete medium, high sugar DMEN, 10% FBS, 4mmol / L glutamine, double antibody, cultivate LLC mouse Lewis lung cancer cell line (purchased from Chinese Academy of Sciences Cell Bank) in 5% CO2 incubator ).

[0124] 2.2. Mouse Modeling

[0125] Male BALB / c mice (provided by the Institute of Model Animals, Nanjing University) were selected and divided into three groups:

[0126] i. Control group: BALB / c mice, inject 100uL normal saline subcutaneously, and give placebo (normal saline) by intragastric administration;

[0127] ii. Experimental group 1: BALB / c mice were g...

Embodiment 3

[0146] Example 3: Rh2 directly down-regulates tumor mouse Treg cells

[0147] First, treat tumor mouse CD4 with ginsenoside Rh2 monomer + T cells; then the expression changes of Treg cells were detected by flow cytometric analysis.

[0148] 1. Cell culture

[0149] LLC mouse lung cancer cells (from the Cell Bank of the Chinese Academy of Sciences) and Renca mouse kidney cancer cells (from the Cell Bank of the Chinese Academy of Sciences) were subcultured, digested and collected for the preparation of mouse tumor models.

[0150] i.LLC mouse lung cancer cells

[0151] Complete medium, high glucose DMEN, 10% FBS, 4mmol / L glutamine, double antibody, in 5% CO 2 Incubate at 37°C in an incubator.

[0152] ii.Renca mouse kidney cancer cells

[0153] Complete medium, high glucose DMEN, 10% FBS, 4mmol / L glutamine, double antibody, in 5% CO 2 Incubate at 37°C in an incubator.

[0154] 2. Mouse Modeling

[0155] BALB / c male mice (provided by the Institute of Model Animals, Nanjin...

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Abstract

The invention discloses application of ginsenoside Rh2 and particularly verifies that ginsenoside can be applied to preparation of inhibitors for inhibiting Treg cells or preparation of compositions for preventing and / or treating tumors with Treg cells increased. The ginsenoside Rh2 is capable of improving balance of T cells in human bodies by inhibition of the Treg cells, autoimmunity can be further enhanced, and accordingly an anti-tumor effect or an immunodeficiency disease improvement effect can be achieved.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and provides the application of ginsenoside Rh2 in inhibiting Treg cells and its preparation method. Specifically, the present invention discloses the method and application of ginsenoside Rh2 to enhance the anti-tumor immune response of T cells and inhibit tumor growth by inhibiting Treg cells, so as to improve the body's immune function. Background technique [0002] Tumor is currently one of the most serious diseases that endanger human health. According to WHO statistics, tumor has become the number one killer threatening human life and health. A lot of manpower, material and financial resources have been invested in the research on the prevention and treatment of malignant tumors at home and abroad, but no fundamental breakthrough has been made so far. Although the combination of traditional chemotherapy, surgery, and radiotherapy has successfully improved the cure rate of variou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/704A61P35/00A23L1/29
Inventor 张辰宇王真高崇昆张栋梁湘樱李红雨
Owner JIANGSU MICROMEDMARK BIOTECH
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