New application of astragalus polysaccharide

An astragalus polysaccharide, a new application technology, applied in the field of biomedicine, can solve problems such as affecting the normal hematopoiesis of bone marrow, and achieve the effect of reducing the formation of DPCs and enhancing the proliferation activity

Inactive Publication Date: 2019-08-30
GANSU UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, once the BM-MSCs are damaged, it will affect the normal hemat

Method used

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  • New application of astragalus polysaccharide
  • New application of astragalus polysaccharide
  • New application of astragalus polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: cell culture and grouping

[0023] 1) Preparation of formaldehyde exposure solution: According to the formaldehyde content (M / V) of 37% and relative molecular weight of 30.03, 0.1194 mL of 37% formaldehyde was added to 99.8804 mL of BM-MSCs special medium to obtain 100 Formaldehyde with a concentration of 16mmol / L was filtered through a 0.22 μm filter membrane, aliquoted, and stored at 4°C. Dilute to 120 μmol / L working solution concentration when used.

[0024] 2) Preparation of astragalus polysaccharide solution: Take 20 mg of astragalus polysaccharide powder (purity ≥ 98%), dissolve in 10 mL of DMEM / F12 medium, and filter through a 0.22 μm microporous filter to make the final concentration of astragalus polysaccharide to 2 mg / mL. Store at 4°C and dilute when used.

[0025] Use BM-MSCs special medium containing 10% fetal bovine serum in 5% CO 2 , 37°C, and a saturated humidity incubator. The logarithmic growth phase cells were trypsinized, inoculated,...

Embodiment 2

[0026] Embodiment 2: MTT method detects cell proliferation activity

[0027] Cell number adjusted to 4×10 3 Each well was inoculated in a 96-well plate, grouped and treated according to Example 1, and then 20 μL of MTT was added and incubated for 4 hours. Discard the supernatant, add DMSO to shake, measure the absorbance (A) value at 490nm with a microplate reader, and calculate the cell proliferation rate of each experimental group according to the formula: (A value of the experimental group / A value of the control group)×100%.

[0028] Such as figure 1 , as shown in Table 1, the results of MTT showed that compared with the control group, the cell proliferation activity of human BM-MSCs exposed to formaldehyde was significantly decreased ( P P <0.01), and when APS 1.0 mg / mL acted, the cell proliferation activity reached its peak.

Embodiment 3

[0029] Embodiment 3: Comet experiment detects DNA fragmentation situation

[0030] Take cells in the logarithmic growth phase, prepare 1% normal melting point agarose gel on a frosted glass slide, then mix the cell suspension with 1% low melting point agarose (volume ratio 1:3), and spread the primer Then, immerse in the lysis solution containing 40% proteinase K, and lyse at 37°C for 4h. Then immerse in 4°C electrophoresis buffer for 30 minutes to unspin. After the unspin is completed, adjust the voltage to 15 V and the current to 300 mA, and avoid electrophoresis for 25 minutes. After electrophoresis, take out the slide, immerse in 0.4 mol / L Tris buffer (pH 7.5), neutralize in the dark at 4°C for 15 min, and then wash with 1% H 2 o 2 Fix, stain with propidium iodide (PI) for 30 min, observe with a fluorescence microscope and take pictures, and analyze the collected pictures by using the comet experiment professional analysis software CASP, and randomly select 20 cells in e...

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Abstract

The invention discloses a new application of astragalus polysaccharide, and belongs to the technical field of biology medicine. A basis is provided for preventing and treating myoblastosis through radix astragali. The inventor of the invention finds that the astragalus polysaccharide can obviously reinforce the proliferation activity of human BM-MSCs in formaldehyde environment, and can reduce DNAfragmentation and DPCs formation of human BM-MSCs induced by formaldehyde so as to have good protection effects. The astragalus polysaccharide can promote DNA injury repair factors of XPA, XPC, ERCC1, RPA1 and RPA2 mRNA and protein expression, and improve DNA damage repair capacity. The inventor finds that the astragalus polysaccharide can reinforce the proliferation activity of human BM-MSCs inthe formaldehyde environment for the first time, can reduce the DNA fragmentation effects of the human BM-MSCs in the formaldehyde environment and can reduce the DPCs formation of the human BM-MSCs inthe formaldehyde environment.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a new application of astragalus polysaccharide. Background technique [0002] As an environmental pollutant, formaldehyde widely exists in automobile exhaust, tobacco smoke and various home decorations. It has obvious toxic effects on the eyes, respiratory, immune, nervous and hematopoietic systems, and can cause cell DNA damage. DNA breakage and DNA-protein crosslinks (DNA-protein crosslinks, DPCs) formation are considered to be the main manifestations of DNA damage caused by formaldehyde. There are even reports that DPCs are molecular markers of formaldehyde-induced carcinogenesis. The higher the level of DPCs, the more cancerous. The higher the risk. Bone marrow mesenchymal stem cells (BM-MSCs) are pluripotent stem cells that exist in the bone marrow. They not only have multilineage differentiation potential, but also regulate the normal proliferation and diff...

Claims

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Application Information

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IPC IPC(8): A61K31/715A61P35/02A61P39/00
CPCA61K31/715A61P35/02A61P39/00
Inventor 舍雅莉李亚玲张立刘永琦李能莲
Owner GANSU UNIV OF CHINESE MEDICINE
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