The use of baicalin combined with Atractylodes polysaccharides to up-regulate Treg cells in the treatment of immune recurrent abortion

A technique for treating Atractylodes polysaccharides and cells, which is applied in the field of medicine

Active Publication Date: 2022-03-11
JIANGSU PROVINCIAL HOSPITAL OF TCM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Studies have shown that baicalin and Atractylodes polysaccharides are the main immune-related pharmacodynamic substance bases of Scutellaria baicalensis and Atractylodes rhizome, respectively, and both have regulatory effects on immune function. However, current research is mainly focused on anti-apoptosis and immune activation. There are few reports on the promotion of immune tolerance in the treatment of immune RSA

Method used

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  • The use of baicalin combined with Atractylodes polysaccharides to up-regulate Treg cells in the treatment of immune recurrent abortion
  • The use of baicalin combined with Atractylodes polysaccharides to up-regulate Treg cells in the treatment of immune recurrent abortion
  • The use of baicalin combined with Atractylodes polysaccharides to up-regulate Treg cells in the treatment of immune recurrent abortion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Detection of the dose-effect relationship between baicalin and Atractylodes macrocephala polysaccharide, such as Figure 9 As shown, (1) Separation of mouse spleen cells followed by red blood cell lysis, removal of adherent cells, magnetic bead sorting of CD4+ T cells; (2) addition of anti-CD3 and anti-CD28 co-stimulatory factors as a blank control group, the experiment On the basis of adding anti-CD3 and anti-CD28, different concentrations of baicalin and Atractylodes polysaccharide were added to induce the differentiation of mouse Treg cells; (3) The concentrations of baicalin were 5 μM, 10 μM, 20 μM and 40 μM; The concentrations were 50 μg / ml, 100 μg / ml, and 200 μg / ml; (4) The proportion of Treg cells in each group of cells was detected by flow cytometry, and a curve was drawn to obtain the dose-effect relationship diagram of baicalin and Atractylodes macrocephala polysaccharide. The experimental results show that: the concentration of baicalin between 5 μ...

Embodiment 2

[0059] Example 2: Baicalin and Atractylodes polysaccharides respectively and jointly promote the flow detection of Treg cells, such as Figure 10 As shown, (1) Preparation of cells for detection: CD4+ T cells sorted by magnetic beads were spread in 24-well plates with 106 cells per well, and different concentrations of Atractylodes polysaccharide RAMP and baicalin were added to each group and combined administration , the cells can be collected after 72 hours for subsequent flow detection; (2) flow antibody staining: centrifuge at 400g for 5 minutes at 4°C, collect the cell pellet and medium supernatant respectively, wash the cell pellet with 1mL pre-cooled PBS buffer 2 times. Add 100 μL of anti-mCD16 / 32 blocking working solution to each tube of cells, incubate in an ice bath in the dark for 20 minutes, and add 1 mL of pre-cooled PBS buffer to wash the cell pellet twice. Add 2 μL FITC anti-mouse CD4 flow cytometry antibody and 5 μL APC anti-mouse CD4 flow cytometry antibody t...

Embodiment 3

[0060] Example 3: Atractylodes polysaccharides and baicalin respectively and jointly promote the qPCR detection of Treg cells, such as Figure 11 Shown, (1) the extraction of RNA: according to Qiangen company's The Plus Mini Kit instructions were used to extract total RNA; (2) RNA reverse transcription: according to Novizan’s The instructions of II Q RT SuperMix for qPCR kit guide to complete the reverse transcription of RNA; (3) Fluorescent real-time quantitative PCR: 1) The fluorescent real-time quantitative PCR (Quantitative real-time PCR, qPCR) primers used in this experiment were provided by Nanjing GenScript Company Synthesis. Centrifuge the synthesized dry powder of primers to make the primers gather at the bottom of the EP tube. Add nuclease-free water to dissolve the primers according to the instructions given by the company, and dilute the concentrated primer stock solution to a concentration of 2uM for use.

[0061]2) Each group needs to set up 3 duplicate wel...

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Abstract

The present invention provides the use of baicalin combined with Atractylodes polysaccharides to up-regulate Treg cells in the treatment of immune recurrent abortion, and relates to the use of baicalin and Atractylodes polysaccharides, including the combination of baicalin and / or Atractylodes polysaccharides to stimulate Naïve CD4 (+) T cells to promote Treg The level of cell expression; and / or; baicalin and / or Atractylodes polysaccharides reduce the embryo absorption rate; the drug combination of baicalin and Atractylodes polysaccharides stimulates Naïve CD4 (+) T cells to promote the expression level of Treg cells, and the present invention provides a The combination of traditional Chinese medicine monomers and polysaccharide compounds, namely baicalin and Atractylodes polysaccharides, verified the effect of the drug combination in increasing immune tolerance, and confirmed its use in the treatment of immune RSA. The present invention uses baicalin and Atractylodes polysaccharide to stimulate Naïve CD4(+) T cells, analyzes the expression level of Treg cells, and determines the optimal dosage ratio of baicalin and Atractylodes polysaccharide for treating immune RSA.

Description

technical field [0001] The invention belongs to the field of medicine and relates to the use of baicalin and Atractylodes polysaccharides, in particular to the use of baicalin and Atractylodes polysaccharides in the preparation of drugs for promoting the proliferation and activation of Treg cells. Background technique [0002] Recurrent spontaneous abortion (RSA) refers to 2 or more consecutive abortions before 20 weeks of pregnancy, which are called "fetal leakage", "fetal movement" and "slip tire" in ancient Chinese medicine books. The incidence of recurrent miscarriage in women of reproductive age is 1% to 5%, of which 50% to 60% are attributed to immune factors. Immune RSA refers to recurrent miscarriages caused by immune rejection between mother and fetus. It is manifested as a lack of immune tolerance. The incidence rate is quite high, and the incidence rate has a tendency to increase year by year, seriously affecting women's reproductive health. [0003] Modern medic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/715A61P15/06A61P37/06A61K31/7048
CPCA61K31/7048A61K31/715A61P15/06A61P37/06A61K2300/00
Inventor 陈聪郭薇陆启滨任青玲康安
Owner JIANGSU PROVINCIAL HOSPITAL OF TCM
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