CN112656807A - Application of platycodon grandiflorum polysaccharide in preparation of medicine for degrading SOCS1/2 - Google Patents
Application of platycodon grandiflorum polysaccharide in preparation of medicine for degrading SOCS1/2 Download PDFInfo
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Abstract
The invention discloses application of platycodon grandiflorum polysaccharide in preparation of a medicine for degrading SOCS1/2, and belongs to the field of medical application of platycodon grandiflorum polysaccharide. The invention discovers for the first time that platycodon grandiflorum polysaccharide with a specific composition can obviously reduce the expression levels of SOCS1 protein and SOCS2 protein in macrophages; further research shows that the platycodon grandiflorum polysaccharide realizes the degradation of SOCS1/2 protein by activating a lysosome pathway. SOCS1/2 may be a logical target for treatment regimens for various immune diseases that occur due to an over-regulation of the immune system, such as immunodeficiency and immunosuppressive diseases. The invention is expected to provide a new therapeutic drug for treating SOCS1/2 immune diseases.
Description
Technical Field
The invention belongs to the field of medical application of platycodon grandiflorum polysaccharide, and particularly relates to application of platycodon grandiflorum polysaccharide in preparation of a medicine for degrading SOCS 1/2.
Background
Cytokine signal transduction inhibitors (SOCS), a class of negative regulators produced by cells and which feedback-block cytokine signaling, regulate a variety of cytokines, and are involved in a variety of inflammatory disease processes in humans.
The SOCS protein family mainly comprises 8 members, SOCS1-7 and CIS. Their protein structures are similar: consists of an N-terminal variable region, a central Src homology region 2(Src homology 2, SH2) tyrosine kinase catalytic function region and a C-terminal SOCS box. The length of the N-terminal region of the SOCS protein is variable, varying from about 50 to about 380 amino acids, and the members of the SOCS family differ in the amino acid sequence and length of this region; the SH2 region contains an SH2 structural domain, can be combined with phosphorylated tyrosine residues of other signal proteins and is a key part for the SOCS protein to play a role; the "SOCS box" is a highly conserved motif consisting of 40 amino acids, with 2-10 non-conserved amino acids in the middle, with homology above 80%. The mechanism of action of SOCS is mainly dependent on the binding capacity of the SH2 domain to the tyrosine phosphorylation site and is additionally associated with the ability of the SOCS cassette to bind to Elongin BC. Overexpression of SOCS in different cell lines has a broad inhibitory effect on cytokine signaling. Research shows that the cell factors activate multiple signal pathways and widely participate in the processes of proliferation, apoptosis, differentiation, development and the like of cells.
In recent years, significant progress has been made in the study of the mechanism by which SOCS inhibits cytokine signaling, and in particular, the negative feedback effect on the JAK/STAT signaling pathway has been demonstrated by several studies. Much of the current research on the SOCS family has focused on SOCS 1-3. Wherein SOCS1 is an important member of a family discovered in the early stage, and researches show that SOCS1 participates in the negative regulation of multiple signal pathways such as JAKs-STAT pathway, NF-kB pathway and the like; in addition, SOCS1 is also closely related to the occurrence of hepatitis, malignant tumors and various autoimmune diseases. SOCS2 can not only regulate the expression of other family members to cause the change of inflammatory factors, but also directly participate in various signal paths to regulate the expression of IFN-gamma, IL-6, IL-12 and other inflammatory factors, and has influence on the balance of Th1/Th 2; it has also been reported that SOCS2 is involved in inflammatory responses and in the accumulation of neutrophils.
However, in the research on SOCS1 and SOCS2, mainly by means of high expression of SOCS1/2 or knockout of SOCS1/2, there are few reports related to direct degradation of SOCS 1/2.
Disclosure of Invention
Aiming at the prior art, the invention aims to provide a new application of platycodon grandiflorum polysaccharide in preparing a medicine for degrading SOCS 1/2.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect of the invention, there is provided the use of platycodon grandiflorum polysaccharide in the manufacture of a medicament for degrading SOCS1 and/or SOCS2 in macrophages.
In the above application, the platycodon grandiflorum polysaccharide degrades SOCS1 and/or SOCS2 by activating lysosomal pathways.
In the application, the macrophage is a porcine alveolar macrophage cell line 3D 4/21.
In the above application, the platycodon grandiflorum polysaccharide is selected from platycodon grandiflorum total polysaccharide (PGPStc) and fractionated Platycodon Grandiflorum Polysaccharide (PGPS)60c) And fractionating Platycodon Grandiflorum Polysaccharide (PGPS)80c) Mixed according to the weight ratio of 2:1: 1.
Preferably, the platycodon grandiflorum total polysaccharide (PGPStc) is prepared by the following method:
soaking radix Platycodi decoction pieces in 8 weight times of water for 30min, boiling for 3 times, each time for 1 hr, mixing filtrates, heating and concentrating the filtrate, cooling, centrifuging at 3000rpm for 10min, and removing impurities to obtain a final concentration of 1 g/mL; slowly adding 95% ethanol until the final concentration of ethanol reaches 80%, stirring while adding, standing for 24 hr, centrifuging, drying the precipitate, and purifying.
The fractionated Platycodon Grandiflorum Polysaccharide (PGPS)60c) And fractionating Platycodon Grandiflorum Polysaccharide (PGPS)80c) The preparation method comprises the following steps:
soaking radix Platycodi decoction pieces in 8 weight times of water for 30min, boiling for 3 times, each time for 1 hr, mixing filtrates, heating and concentrating the filtrate, cooling, centrifuging at 3000rpm for 10min, and removing impurities to obtain a final concentration of 1 g/mL; slowly adding 95% ethanol until the final concentration of ethanol reaches 60%, stirring, standing for 24 hr, centrifuging, drying the precipitate, and purifying to obtain graded radix Platycodi polysaccharide (PGPS)60c)；
Slowly adding 95% ethanol into the centrifuged supernatant until the final concentration of ethanol reaches 80%, stirring while adding, standing for 24 hr, centrifuging, drying the precipitate, and purifying to obtain graded radix Platycodi polysaccharide (PGPS)80c)。
Preferably, the fractionated Platycodon Grandiflorum Polysaccharide (PGPS)60c) In the formula, the molar ratio of glucose is 62.603%, and the molar ratio of arabinose is 62.603%13.925%; the fractionated Platycodon Grandiflorum Polysaccharide (PGPS)80c) In the formula, the molar ratio of glucose is 54.322%, the molar ratio of arabinose is 16.801%, and the molar ratio of galactose is 15.013%.
Preferably, the fractionated Platycodon Grandiflorum Polysaccharide (PGPS)60c) Has a number average molecular weight of 3.00X 103-9.34×104Da, weight average molecular weight 5.69X 103-1.12×105Da, distribution width is 1.2-1.89; the fractionated Platycodon Grandiflorum Polysaccharide (PGPS)80c) Has a number average molecular weight of 3.13X 103-6.12×104Da with a weight average molecular weight of 4.14X 103-1.01×105Da, distribution width 1.32-1.64.
In a second aspect of the invention, a medicament for degrading SOCS1 and/or SOCS2 in macrophages is provided, wherein the medicament takes platycodon grandiflorum polysaccharide as an effective component; the radix Platycodi polysaccharide is prepared from total polysaccharide (PGPStc) and fractionated polysaccharide (PGPS)60c) And fractionating Platycodon Grandiflorum Polysaccharide (PGPS)80c) Mixed according to the weight ratio of 2:1: 1.
The invention has the beneficial effects that:
the invention discovers for the first time that platycodon grandiflorum polysaccharide with a specific composition can obviously reduce the expression levels of SOCS1 protein and SOCS2 protein in macrophages; further research shows that the platycodon grandiflorum polysaccharide realizes the degradation of SOCS1/2 protein by activating a lysosome pathway. SOCS1/2 may be a logical target for treatment regimens for various immune diseases that occur due to an over-regulation of the immune system, such as immunodeficiency and immunosuppressive diseases. The invention is expected to provide a new therapeutic drug for treating SOCS1/2 immune diseases.
Drawings
FIG. 1: LPS/IFN-gamma induces the morphological change of the cells after M1 polarization of 3D4/21 cells; in the figure, A is CON group and B is IFN-. gamma. + LPS treated group.
FIG. 2: mRNA expression of SOCS1/2 after LPS/IFN-. gamma.and PGPS treatment; in the figure, A is the expression of SOCS1 mRNA; b is SOCS2mRNA expression.
FIG. 3: protein levels of SOCS1/2 were varied after LPS/IFN-. gamma.and PGPS treatment.
FIG. 4: expression of iNOS after LPS/IFN-. gamma.and PGPS treatment.
FIG. 5: protein levels of PGPS-treated SOCS1/2 changed after LPS/IFN-. gamma.and MG-132 or CQ.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
As described in the background section, SOCS1 and SOCS2 are involved in the regulation of multiple signaling pathways and are also closely related to the development of various autoimmune diseases. Therefore, the development of corresponding regulatory substances by taking SOCS1/2 as a target is of great significance for the treatment of immune diseases.
Platycodon grandiflorum is one of the most representative Chinese medicines in China, and has a history of hundreds of years in China. Radix Platycodi can promote health and homeostasis. In recent decades, the research on platycodon grandiflorum has mainly focused on its biological activities such as anti-tumor, liver protection, immunoregulation and anti-oxidation. These studies have isolated compounds such as saponins, flavonoids, anthocyanins, phenols and polysaccharides from plants. The inventor of the present patent conducted various studies on extraction, structural identification and function of platycodon grandiflorum polysaccharide in the early stage. During the development process of the SOCS1/2 regulating substance, the invention unexpectedly discovers that the expression level of SOCS1/2 protein is increased by LPS/IFN-gamma, but the protein level of SOCS1/2 is reduced by platycodon grandiflorum polysaccharide.
On the basis, the invention further researches the degradation pathway of the platycodon grandiflorum polysaccharide to SOCS1/2, and as a result, the platycodon grandiflorum polysaccharide is found to degrade SOCS1/2 by activating a lysosome pathway.
In addition, the platycodon grandiflorum polysaccharide prepared by different methods has different chemical compositions and molecular weights; the type and molar ratio of monosaccharides in platycodon grandiflorum polysaccharide are closely related to the biological activity of polysaccharide. In order to further research the influence of the composition of the platycodon grandiflorum polysaccharide on the degradation effect of SOCS1/2, the invention uses the identification of the structure of the platycodon grandiflorum polysaccharide and the influence on the immunocompetence of chicken abdominal cavity macrophages (Shandong agriculture Dada)Radix Platycodi total polysaccharide (PGPStc) and fractionated radix Platycodi polysaccharide (PGPS) prepared in academic thesis of Master school, 2017)60c) And fractionating Platycodon Grandiflorum Polysaccharide (PGPS)80c) For the test material, the effect of the test material on the degradation effect of SOCS1/2 in macrophage 3D4/21 alone and after mixing was examined, and the results are as follows:
the above results indicate that the total polysaccharide (PGPStc) and fractionated polysaccharide (PGPS) of Platycodon grandiflorum60c) And fractionating Platycodon Grandiflorum Polysaccharide (PGPS)80c) The compound has the optimal effect on the degradation of SOCS1/2 in macrophage 3D4/21 and has a remarkable synergistic promoting effect.
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
The test materials used in the examples of the present invention, which were not specifically described, were all those conventional in the art and commercially available. Wherein:
the platycodon grandiflorum polysaccharide used in the present example is composed of platycodon grandiflorum total polysaccharide (PGPStc) and fractionated Platycodon Grandiflorum Polysaccharide (PGPS)60c) And fractionating Platycodon Grandiflorum Polysaccharide (PGPS)80c) Mixed in a weight ratio of 2:1:1 and marked as PGPSt. Radix Platycodi total polysaccharide (PGPStc), and fractionated radix Platycodi polysaccharide (PGPS)60c) And fractionating Platycodon Grandiflorum Polysaccharide (PGPS)80c) Prepared according to the structural identification of the platycodon grandiflorum polysaccharide and the influence on the immunological activity of chicken abdominal cavity macrophages (Master academic paper of Shandong university of agriculture, 2017), namely 'extraction of 2.2.1 polysaccharide' and 'purification of 2.2.2 polysaccharide'.
Recombinant porcine interferon gamma (IFN-. gamma.) (985-PI050) was purchased from R-D Systems (Minneapolis, MN, USA). Lipopolysaccharide (LPS) (L8880) was purchased from Solarbio (Beijing, China). MG-132, Chloroquine (CQ), supplied by MCE corporation (Shanghai, China). hoechst33342 kit (C1026) and nitric oxide synthase (iNOS) detection kit (S0025) were provided by the Beyotime Biotechnology institute (Haima, Jiangsu, China). Modified RPMI-1640 medium was purchased from Gibco (Shanghai, China). Penicillin streptomycin amphotericin B (03-033-1B/C) and certified FBS (04-001-1ACS) were purchased from Kibbutz wait Haemek Biotech, Israel.
Antibody: anti-SOCS1(ab9870), available from Abcam (Shanghai, China); anti-SOCS2(#2779) was purchased from CST (Shanghai, China); anti-GAPDH (600041lg), HRP-conjugated affinity rabbit anti-goat IgG (H L) (SA00001-4), HRP-conjugated goat anti-mouse IgG (H L) (SA00001-1), HRP-conjugated goat anti-rabbit IgG (H L) (SA00001-2) and CoraLite 488-conjugated goat anti-mouse IgG (H L) (SA00013-1) were purchased from Proteintetech, Inc., Wuhan, China.
The porcine alveolar macrophage cell line 3D4/21 was purchased from iCell Bioscience (Shanghai, China).
Example 1: research on influence of platycodon grandiflorum polysaccharide on expression of SOCS1/2 in porcine alveolar macrophages
1. The test method comprises the following steps:
1.1, preparing a platycodon grandiflorum polysaccharide solution:
the platycodon grandiflorum polysaccharide prepared in example 1 was prepared into a PGPSt solution with a concentration of 100. mu.g/ml using RPMI 1640 medium.
1.2 cell culture:
the porcine alveolar macrophage cell line 3D4/21 was cultured in modified RPMI-1640 medium containing 2mM L-glutamine, 1.5g/L sodium bicarbonate, 4.5g/L glucose, 10mM HEPES, 1.0mM sodium pyruvate, with the addition of 0.1mM non-essential amino acids, 10% fetal bovine serum, 1% penicillin-mycin amphotericin B; culturing in 5% carbon dioxide at 37 deg.C.
1.3 cell treatment:
dividing the cultured porcine alveolar macrophages into three groups, and using a CON group as a blank control without treatment; IFN-gamma + LPS treated groups stimulated porcine alveolar macrophages with LPS (150ng/ml) and IFN-gamma (50 g/ml); IFN-. gamma. + LPS + PGPSt-treated groups porcine alveolar macrophages were stimulated with LPS (150ng/mL) and IFN-. gamma. (50g/mL) and grown to 70-80% in 6-well plates, after which PGPSt solution (100. mu.g/mL) was added. After various treatments, cells or supernatants were collected for analysis.
1.4 real-time fluorescent quantitative Polymerase Chain Reaction (PCR)
Total RNA was extracted with RNAiso Plus regent (Takara, Dalian) and genomic DNA was removed using PrimeScriptfi RT kit (Takara, Dalian) containing a gDNA eraser and reverse transcribed in a BIO-RAD Mastercycler (Takara, Dalian). PCR was performed in triplicate in the LightCyclerfi96 system (roche, germany). The sequences of the specific primers are shown in Table 1. Foldchange was calculated after normalizing the change in expression of the housekeeping gene GAPDH gene using the threshold cycle value.
Table 1: specific primer sequence
1.5 protein isolation and Western blotting
After the different treatments, the cells were protein-separated with RIPA buffer containing a mixture of protease inhibitors. Protein concentration was determined using BCA protein assay kit (CWBIO, beijing). Equivalent protein samples were size separated by 12% SDS-PAGE and blotted with polyvinylidene fluoride membrane. After blocking, the cell membrane was exposed overnight to antibodies recognizing SOCS1, SOCS 2. The IgG secondary antibody was incubated with the corresponding horseradish peroxidase (HRP) conjugated IgG for 1h at room temperature, the reaction bands were visualized with an enhanced chemiluminescence detection system and analyzed with Image J software. Relative protein expression levels were normalized to GAPDH.
1.6 nitric oxide synthase assays
And detecting the expression condition of the nitric oxide synthase under different treatments by adopting a nitric oxide synthase (iNOS) detection kit.
1.7 statistical analysis
All data were from triplicate experiments and are expressed as Standard Deviation (SD). Differences between groups were analyzed using one-way analysis of variance (ANOVA). The significance levels were as follows: p- <0.05 and P- <0.01 are indicated by x.
2. And (3) test results:
2.1 morphological Change in M1 polarization in porcine alveolar macrophages
After 24 hours of stimulation with LPS/IFN- γ, the morphology of 3D4/21 cells changed. Compared with CON (FIG. 1A), the nucleus of the polarized 3D4/21 cell becomes larger, the nucleus becomes irregular, the cell contour becomes round, and tubulin becomes disorganized (FIG. 1B).
2.2 mRNA expression level of SOCS1/2 in porcine alveolar macrophages
As shown in FIGS. 2A and B, LPS/IFN- γ also enhanced the mRNA expression level of SOCS 1/2; however, the addition of PGPSt did not significantly affect the mRNA expression level of SOCS 1/2.
2.3 protein levels of SOCS1/2 in porcine alveolar macrophages
LPS/IFN-gamma treatment increased the expression level of SOCS1/2 protein in macrophages; however, the addition of PGPSt resulted in a significant decrease in the protein level of SOCS1/2 (FIGS. 3A, B and C).
2.4 expression of iNOS in porcine alveolar macrophages
The expression of nitric oxide synthase under different treatments is shown in FIG. 4, and the increase of the content of nitric oxide synthase indicates the increase of M1 polarization of macrophage.
The above test results show that: PGPSt was able to significantly reduce the increase in SOCS1/2 protein expression levels in macrophages caused by LPS/IFN- γ treatment; however, PGPSt did not have a significant effect on the mRNA expression level of SOCS1/2, and thus it was speculated that PGPSt reduced the expression level of SOCS1/2 protein in macrophages by degradation.
Example 2: research on degradation pathway of platycodon grandiflorum polysaccharide to SOCS1/2 in porcine alveolar macrophages
To explore the degradation pathway of SOCS1/2, the present invention used the classical proteasome pathway inhibitor MG-132 and the lysosomal inhibitor CQ. Performing cell culture according to the method of example 1, dividing cultured porcine alveolar macrophages into five groups, and using CON group as blank control without treatment; IFN-gamma + LPS treated groups stimulated porcine alveolar macrophages with LPS (150ng/ml) and IFN-gamma (50 g/ml); IFN- γ + LPS + PGPSt treated groups porcine alveolar macrophages were stimulated with LPS (150ng/mL) and IFN- γ (50g/mL) to 70-80% growth in 6-well plates, and then PGPSt solution (100. mu.g/mL) was added; IFN- γ + LPS + PGPSt + MG-132 treatment groups porcine alveolar macrophages were stimulated with LPS (150ng/mL) and IFN- γ (50g/mL) to 70-80% growth in 6-well plates, after which PGPSt solution (100. mu.g/mL) and MG-132(100nM) were added; IFN-. gamma. + LPS + PGPSt + CQ treated groups porcine alveolar macrophages were stimulated with LPS (150ng/mL) and IFN-. gamma. (50g/mL) to grow to 70-80% in 6-well plates, and then PGPSt solution (100. mu.g/mL) and CQ (50. mu.M) were added.
The expression levels of SOCS1 and SOCS2 proteins were determined in the cells after different treatments using the method of "1.5 protein isolation and Western blotting" in example 1, and the results are shown in FIG. 5.
As shown in FIGS. 5A, B and C (it should be noted that, since the optimal exposure intensities of the experimental bands of different batches are automatically analyzed by the instrument, the obtained bands have different differences when subjected to gray scale analysis, but each protein has the internal reference GAPDH from the same gel for comparison, so that the difference between groups is reliable), the expression level of SOCS1/2 protein is reduced by PGPSt, and the expression of SOCS1/2 can still be reduced by PGPSt when treated with MG-132. In contrast, CQ prevented the effect of PGPSt on SOCS 1/2. These results indicate that PGPSt can degrade SOCS1/2 protein by activating the lysosomal pathway.
In sum, macrophages polarize towards classically activated M1 phenotype macrophages and can trigger an inflammatory response and kill pathogens in cells. The IFN- γ/JAK-STAT1 pathway is a key component of macrophage M1 polarization regulators. The SOCS protein family is an important regulator of macrophage polarization. During viral infection, cytokines trigger and manage inflammation. However, excessive cytokine production can cause cytokine storm, and excessive host innate immune response can also damage the body. Therefore, SOCS proteins having negative feedback regulation ability can prevent damage to the host due to excessive secretion of cytokines. For M1-type macrophages, SOCS1/2 protein acts as a feedback inhibitor protein that attenuates the secretion of certain pro-inflammatory mediators by M1-type macrophages. However, studies have shown that overexpression of SOCS1 results in decreased phosphorylation levels of JAK1, TYK2 and STAT1 and inhibits IFN-I-induced antiviral and antiproliferative responses, and that infectious microorganisms can manipulate the SOCS proteins of a host to evade innate immunity, such as Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) which can select SOCS1 to evade the host's immune response, SOCS1 which can inhibit the expression levels of ISGs and IFN- β to promote viral replication, and in addition, immune evasion of various viruses, such as HBV, HCV, Human Immunodeficiency Virus (HIV), Semiki forest, Coxsackie virus, RSV, Ebola virus, IAV, HSV-1, varicella virus (VZV), Japanese Encephalitis Virus (JEV) and Epstein-Barr virus (EBV), can hijack the SOCS proteins to evade innate immunity. Thus, upon infection with certain viruses, an increase in SOCS is detrimental.
The research of the invention finds that the platycodon grandiflorum polysaccharide can degrade SOCS1/2 protein through activating a lysosome way, and the platycodon grandiflorum polysaccharide has very important significance for realizing the control of viruses such as porcine reproductive and respiratory syndrome virus, HBV, HCV, Human Immunodeficiency Virus (HIV), Semliki forest virus, Coxsackie virus, RSV, Ebola virus, IAV, HSV-1, varicella virus (VZV), Japanese Encephalitis Virus (JEV) and Epstein-Barr virus (EBV) by adjusting the level of SOCS1/2 protein.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
SEQUENCE LISTING
<110> Shandongdxin Biotechnology, Inc.; shandong Xingu health industry Co Ltd
Application of <120> platycodon grandiflorum polysaccharide in preparation of medicine for degrading SOCS1/2
<130> 2021
<160> 6
<170> PatentIn version 3.5
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ggcaccgttc acctctatct g 21
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Claims (8)
1. Use of platycodon grandiflorum polysaccharide in the preparation of a medicament for degrading SOCS1 and/or SOCS2 in macrophages.
2. The use according to claim 1, wherein the platycodon grandiflorum polysaccharide degrades SOCS1 and/or SOCS2 by activating lysosomal pathways.
3. The use of claim 1, wherein the macrophage is porcine alveolar macrophage cell line 3D 4/21.
4. The use as claimed in claim 1, wherein the platycodon grandiflorum polysaccharide is selected from the group consisting of PGPStc, PGPS60cAnd PGPS80cMixed according to the weight ratio of 2:1: 1.
5. The use according to claim 4, wherein the PGPStc is prepared by a process comprising:
soaking radix Platycodi decoction pieces in 8 weight times of water for 30min, boiling for 3 times, each time for 1 hr, mixing filtrates, heating and concentrating the filtrate, cooling, centrifuging at 3000rpm for 10min, and removing impurities to obtain a final concentration of 1 g/mL; slowly adding 95% ethanol until the final concentration of ethanol reaches 80%, stirring while adding, standing for 24 hr, centrifuging, drying the precipitate, and purifying to obtain the final product;
the PGPS60cAnd PGPS80cThe preparation method comprises the following steps:
soaking radix Platycodi decoction pieces in 8 weight times of water for 30min, boiling for 3 times, each time for 1 hr, mixing filtrates, heating and concentrating the filtrate, cooling, centrifuging at 3000rpm for 10min, and removing impurities to obtain a final concentration of 1 g/mL; slowly adding 95% ethanol until the final concentration of ethanol reaches 60%, stirring, standing for 24 hr, centrifuging, drying the precipitate, and purifying to obtain PGPS60c；
Slowly adding 95% ethanol into the centrifuged supernatant until the final concentration of ethanol reaches 80%, stirring while adding, standing for 24 hr, centrifuging, drying the precipitate, and purifying to obtain PGPS80c。
6. The use according to claim 4, wherein the PGPS is selected from the group consisting of60cIn the formula, the molar ratio of glucose is 62.603%, and the molar ratio of arabinose is 13.925%; the PGPS80cIn the formula, the molar ratio of glucose is 54.322%, the molar ratio of arabinose is 16.801%, and the molar ratio of galactose is 15.013%.
7. The use according to claim 4, wherein the PGPS is selected from the group consisting of60cHas a number average molecular weight of 3.00X 103-9.34×104Da, weight average molecular weight 5.69×103-1.12×105Da, distribution width is 1.2-1.89; the PGPS80cHas a number average molecular weight of 3.13X 103-6.12×104Da with a weight average molecular weight of 4.14X 103-1.01×105Da, distribution width 1.32-1.64.
8. A medicine for degrading SOCS1 and/or SOCS2 in macrophages is characterized in that the medicine takes platycodon grandiflorum polysaccharide as an effective component; the platycodon grandiflorum polysaccharide is composed of PGPStc and PGPS60cAnd PGPS80cMixed according to the weight ratio of 2:1: 1.
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Application publication date: 20210416 |