CN116693707A - Russula cinerea polysaccharide with hematopoiesis promoting effect and preparation method and application thereof - Google Patents
Russula cinerea polysaccharide with hematopoiesis promoting effect and preparation method and application thereof Download PDFInfo
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- CN116693707A CN116693707A CN202310055533.9A CN202310055533A CN116693707A CN 116693707 A CN116693707 A CN 116693707A CN 202310055533 A CN202310055533 A CN 202310055533A CN 116693707 A CN116693707 A CN 116693707A
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- Prior art keywords
- polysaccharide
- russula
- rgp
- hematopoiesis
- promoting effect
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Abstract
The invention belongs to the technical field of medicines, and particularly discloses a russula cinerea polysaccharide with hematopoiesis promoting effect, a preparation method and application thereof. Further experiments show that the active polysaccharide component can increase CD4 in mice with hematopoietic dysfunction caused by chemotherapy + The expression of T cells promotes the recovery of the hematopoietic function of the organism, and has good hematopoietic function promotionThe method has great significance for the research and development of the russula polysaccharide functional food.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to russula gray mushroom polysaccharide with hematopoiesis promoting effect, and a preparation method and application thereof.
Background
In mammals, hematopoietic Stem Cells (HSCs), which originate in the yolk sac of the embryo and in fetuses and adults, are capable of constantly self-renewing and differentiating into various blood cells, including erythrocytes, lymphocytes, platelets, etc., ensuring steady-state production of hematopoietic cells and a lifelong immune response. The homeostasis of HSCs is affected by the microenvironment in which they are located, i.e. the stem cell niche, which can be divided into two classes, osteoblast (endosteal) niches and vascular (sinusoidal) niches, depending on their cellular compartments, which support hematopoietic stem/progenitor cell homing, self-renewal regulation and differentiation of cells of the hematopoietic lineage. The stromal cell fraction of the niche can produce a variety of cytokines, such as Stem Cell Factor (SCF), interleukin (IL) -7, and CXC chemokine ligand (CXCL) 12, which are necessary for HSCs to maintain hematopoietic progression.
Currently, chemotherapy in cancer treatment is considered as a major method for combating diseases, and cell inhibitors such as cyclophosphamide, cisplatin and doxorubicin, which not only exhibit toxic effects on cancer cells, but also inhibit proliferation of blood cells in bone marrow, and even cause hematopoietic injury diseases such as acute neutropenia, lymphopenia, erythropenia, thrombocytopenia, etc. The existing treatment means for relieving hematopoietic injury mainly comprise administration of recombinant granulocyte colony stimulating factor, thrombopoietin, constitutive blood transfusion, and treatment of natural products. The polysaccharide is a natural product of a macromolecule, has the characteristics of multiple targets, strong pharmacological action and small toxic and side effects, has been reported to have the effect of stimulating hematopoiesis, and can be used as a therapeutic agent or an auxiliary therapeutic agent for regulating the hematopoiesis function of organisms.
The russula vinosa is a medicinal and edible wild fungus, and the fungus cover is red-purple and widely distributed in the southern area of China. The russula vinosa has delicious taste, can be used as a main component of functional foods, and has proved to have various pharmacological effects such as immunity enhancement, anti-inflammatory, antioxidation and the like. The Riche griseus has rich pharmacological activity and biological active substances and rich nutritional ingredients, such as high protein, high polysaccharide content and low fat content. At present, the effect of the russula gray polysaccharide component on relieving the hematopoietic injury of the organism has not been studied and reported.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a russula polysaccharide with hematopoiesis promoting effect and a preparation method and application thereof.
In order to achieve the above purpose, the invention is implemented according to the following technical scheme:
the first object of the present invention is to provide a russula gray polysaccharide with hematopoiesis promoting effect, which has a molecular weight of 31.4kDa and is mainly composed of a molar mass ratio of 25.77:8.37:2.51:1.52: galactose, methoxy galactose, glucose, mannose, fucose of 1.41.
The second object of the invention is to provide a preparation method of russula polysaccharide with hematopoiesis promoting effect, which comprises the following steps:
s1, taking and drying the russula vinosa fruiting bodies, and crushing the fruiting bodies by using a crusher;
s2, according to the mass of the russula vinosa fruiting bodies: deionized water volume = 1: mixing at 30-50 ratio, leaching with hot water at 60-90deg.C for 2-4 hr; repeating the extraction twice, combining the two extracting solutions, and concentrating the extracting solution by rotary evaporation;
s3, removing protein components in the extracting solution by using a Sevag method, discarding organic reagents, and performing rotary evaporation and dialysis on the upper extracting solution;
s4, adding an absolute ethanol solution into the dialyzed liquid to ensure that the final concentration of the ethanol is 80%, fully and uniformly mixing, standing for 12 hours at 4 ℃, centrifuging for 20 minutes at 6000rpm after standing, collecting precipitate, drying the ethanol, and freeze-drying to obtain the russula vinosa Linne crude polysaccharide with hematopoiesis promoting effect;
s5, dissolving the russula botrytis cinerea crude polysaccharide by using ultrapure water, wherein the concentration of the prepared polysaccharide solution is 0.05-0.1g/mL, passing through a 0.45um filter membrane after the polysaccharide solution is fully dissolved, loading the solution on a DEAE Sepharose FF ion exchange column, eluting mobile phases respectively into 0, 0.1 and 0.3M NaCl solutions, sequentially named RGP-A, RGP-B, RGP-C as polysaccharide components eluted by the mobile phases, and collecting 7mL of liquid per tube at the flow rate of 1-2 mL/min; measuring polysaccharide content in the collected liquid by adopting a phenol-sulfuric acid method, drawing an elution curve, collecting a polysaccharide tube with higher concentration, performing rotary evaporation, dialysis and freeze-drying;
s6, separating and purifying by using a DEAE chromatographic column to finally obtain two main polysaccharide components, namely RGP-A, RGP-B; using K562 cells, combining with an XTT method, an apoptosis detection method, a benzidine staining method and a western blot method to analyze and screen RGP-A with better in-vitro hematopoietic promotion activity to be applied to subsequent purification;
s7, sequentially loading RGP-A on HiPrepTM 26/60SephacrylTM S-400 and Ezload26/60Chromdex 200pg gel columns, eluting the mobile phase with 0.15M NaCl, and collecting 2-4mL of liquid in each tube at a flow rate of 1 mL/min; and (3) measuring the polysaccharide content in the collected liquid by adopting a phenol-sulfuric acid method, drawing an elution curve, collecting a polysaccharide tube with higher concentration, performing rotary evaporation, dialysis and freeze-drying to obtain the purified polysaccharide of the russula cinerea with hematopoiesis promoting effect.
The third object of the invention is to provide an application of russula vinosa polysaccharides with hematopoiesis promoting effect in preparing medicines for promoting hematopoiesis.
Further, the russula vinosa polysaccharides with hematopoiesis promoting effect are used for preparing medicines for relieving hematopoietic dysfunction caused by chemotherapy.
Compared with the prior art, the invention takes the russula cinerea polysaccharide extracted by a hot water leaching method as a research object, deeply defines the chemical structure of the purified polysaccharide through a series of separation and purification, and explores the activity and action mechanism of the polysaccharide for relieving hematopoietic dysfunction caused by chemotherapy on the basis. The invention has simple operation, easily obtained raw materials and no pollution hazard, can simultaneously determine the structure and the drug effect of the active polysaccharide in the russula vinosa, and has the potential of being developed into a hematopoietic functional product.
Drawings
FIG. 1 is a DEAE-52Sepharose FF elution profile of russula vinosa crude polysaccharide;
FIG. 2 is a HiPrepTM 26/60SephacrylTM S-400 elution profile of Rumex Gracilis polysaccharide RGP-A;
FIG. 3 is an Ezload26/60chromdex 200pg elution profile of russula polysaccharide RGP-A1;
FIG. 4 is a GPC molecular weight measurement spectrum of Rumex Gracilis polysaccharide RGP 1;
FIG. 5 is a monosaccharide composition HPLC diagram of russula polysaccharide RGP 1;
FIG. 6 is an infrared spectroscopic analysis of russula polysaccharide RGP 1;
FIG. 7 is NMR of Rumex Gracilis polysaccharide RGP1 1 H spectrogram;
FIG. 8 is NMR of Rumex Gracilis polysaccharide RGP1 13 C, spectrogram;
FIG. 9 is an NMR COSY spectrum of russula polysaccharide RGP 1;
FIG. 10 is an NMR HSQC spectrum of russula polysaccharide RGP 1;
FIG. 11 is an NMR NOESY spectrum of Rumex Gracilis polysaccharide RGP 1;
FIG. 12 is an NMR HMBC spectra of russula polysaccharide RGP 1;
FIG. 13 is a graph showing the effect of russula polysaccharide RGP1 on the number of hematopoietic-related cells in hematopoietic dysfunction mice;
FIG. 14 is a pathological effect of russula polysaccharide RGP1 on hematopoietic dysfunction mouse bone marrow;
FIG. 15 is a proteomic analysis of russula polysaccharide RGP1 on the spleen of hematopoietic dysfunction mice. (a) thermogram composition analysis of 16 proteins; (B) a STRING network analysis of 16 proteins; (C) GO and (D) KEGG bioinformatics analysis;
FIG. 16 is the effect of russula gray polysaccharide RGP1 on spleen and serum IL-2 levels in hematopoietic dysfunction mice;
FIG. 17 is the effect of russula gray polysaccharide RGP1 on spleen and serum IL-3 levels in hematopoietic dysfunction mice;
FIG. 18 is the effect of russula gray polysaccharide RGP1 on spleen and serum IL-5 levels in hematopoietic dysfunction mice;
FIG. 19 is the effect of russula gray polysaccharide RGP1 on spleen and serum IL-6 levels in hematopoietic dysfunction mice;
FIG. 20 is the effect of russula gray polysaccharide RGP1 on spleen and serum TNF- α levels in hematopoietic dysfunction mice;
FIG. 21 is the effect of russula greysari polysaccharide RGP1 on spleen and serum TGF-beta levels in hematopoietic dysfunction mice.
Detailed Description
The present invention will be described in further detail with reference to the following examples in order to make the objects, technical solutions and advantages of the present invention more apparent. The specific embodiments described herein are for purposes of illustration only and are not intended to limit the invention.
Example 1
Extraction and purification of russula polysaccharide
(1) Drying the russula vinosa fruiting bodies, and crushing the fruiting bodies by using a crusher;
(2) According to the mass of the fruit bodies of the russula vinosa: deionized water volume = 1: mixing the russula vinosa fruiting body with deionized water at a ratio of 30-50, and leaching with hot water at 60-90deg.C for 2-4 hr. Repeating the extraction twice, combining the extracting solutions, and concentrating the extracting solution by rotary evaporation;
(3) Removing protein component in the extractive solution by Sevag method, discarding organic reagent, and further steaming and dialyzing the upper extractive solution;
(4) Adding absolute ethanol solution into the dialyzed liquid to ensure that the final concentration of ethanol is 80%, fully and uniformly mixing, standing for 12 hours at 4 ℃, standing, centrifuging at 6000rpm for 20 minutes, collecting precipitate, drying the ethanol, and freeze-drying to obtain the russula vinosa crude polysaccharide;
(5) The russula botrytis cinerea crude polysaccharide is dissolved by using ultrapure water, the concentration of the prepared polysaccharide solution is 0.05-0.1g/mL, the solution is fully dissolved and then passes through a 0.45um filter membrane, the solution is loaded on a DEAE Sepharose FF ion exchange column, the eluting mobile phases are respectively 0, 0.1 and 0.3M NaCl solution, the polysaccharide components eluted by the mobile phases are sequentially named as RGP-A, RGP-B, RGP-C, the flow rate is 1-2mL/min, and 7mL of liquid is collected per tube. Measuring polysaccharide content in the collected liquid by adopting a phenol-sulfuric acid method, drawing an elution curve, collecting a polysaccharide tube with higher concentration, performing rotary evaporation, dialysis and freeze-drying;
(6) Separating and purifying by DEAE chromatographic column to obtain two main polysaccharide components (RGP-A, RGP-B); using K562 cells, combining with an XTT method, an apoptosis detection method, a benzidine staining method and a western blot method to analyze and screen RGP-A with better in-vitro hematopoietic promotion activity to be applied to subsequent purification;
(7) RGP-A was sequentially loaded onto HiPrep TM 26/60Sephacryl TM S-400 and Ezload26/60Chromdex 200pg gel columns, eluting mobile phase 0.15M NaCl, flow rate 1mL/min, 2-4mL of liquid per tube were collected. And (3) measuring the polysaccharide content in the collected liquid by adopting a phenol-sulfuric acid method, drawing an elution curve, collecting a polysaccharide tube with higher concentration, performing rotary evaporation, dialysis and freeze-drying to obtain the russula vinosa purified polysaccharide component.
Results: the elution result of the ion exchange column chromatography shows that the russula polysaccharide mainly contains two polysaccharide components, namely RGP-A and RGP-B; RGP-A is eluted by further gel column chromatography to finally obtain single polysaccharide component, and the results are shown in figures 1-3.
Example 2
Characterization of the Rumex Gracilis polysaccharide prepared in example 1
(1) Test materials: the russula gray polysaccharide purified in example 1;
(2) Test method and results
Molecular weight measurement: 2mg/mL of russula polysaccharide was loaded onto a Gel Permeation Chromatograph (GPC) equipped with a PL aquagel-OH Mixed-H (7.5 mm. Times.300 mm) column. The eluting mobile phase was a 0.1M sodium nitrate solution containing 0.01% sodium azide and the molecular weight was calculated by fitting the peak time and the sample viscosity.
Molecular weight measurement results: the MP of the russula polysaccharide is 31.547kDa, the Mn is 31.405kDa, the Mw is 31.425kDa, the Mz is 31.4475 kDa, the Mz+1 is 31.463kDa, and the GPC spectrogram shows that the molecular weight of the russula polysaccharide is single peak, which shows that the molecular weight uniformity of the sample is good, and the result is shown in figure 4.
Monosaccharide composition analysis: mixing 5mg of russula polysaccharide with 2M trifluoroacetic acid, and hydrolyzing at 110 ℃ for 2h, followed by pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP); loading standard yeast and derived polysaccharide into preparation C 18 In a high performance liquid chromatograph of a chromatographic column (4.6 mm multiplied by 250 mm), the composition of monosaccharides in the russula polysaccharide hydrolysate is analyzed according to the concentration, the peak time and the peak area of a standard substance.
Monosaccharide composition analysis results: the HPLC comparison calculation of the control standard substance and the russula polysaccharide shows that the russula polysaccharide mainly comprises five monosaccharides of galactose, methoxy galactose, glucose, mannose and fucose, and the molar mass ratio of the monosaccharides is 25.77:8.37:2.51:1.52:1.41, the results are shown in FIG. 5.
And (3) infrared spectrum analysis: 2mg of russula polysaccharide RGP1 and 200mg of potassium bromide are uniformly mixed, pressed into tablets with the thickness of 1mm, and placed in a Nicolet iZ-10 Fourier transform infrared spectrometer for analysis. Scanning with infrared light source with a scanning range of 4000-400cm -1 The number of scans was 32 and the detector was DTGS KBr.
Results of infrared spectroscopy analysis: the infrared analysis of the characteristic absorption peak of the russula cinerea polysaccharide shows that the russula cinerea polysaccharide is 3436.33cm -1 The characteristic peak of the O-H stretching vibration absorption saccharide exists at 2928.92cm -1 There is C-H stretching vibration peak at 1643.85cm -1 There is C=O stretching vibration peak, 1081.48cm -1 There is a C-O stretching vibration peak, and the result is shown in FIG. 6.
Methylation analysis: weighing 1mg of russula polysaccharide, mixing with 500 mu L of DMSO solution, dissolving, sequentially adding 1mg of sodium hydroxide, incubating for 0.5 hour, incubating with 50 mu L of methyl iodide solution for 1 hour, and mixing with 1mL of ultrapure water and 2mL of dichloromethane solution; next, the dichloromethane layer was evaporated to dryness and reacted with 2M trifluoroacetic acid for 1.5 hours, and then evaporated to dryness and reacted with 2M ammoniaWater, 1M NaBD 4 Mixing and reacting for 2.5 hours, and then adding glacial acetic acid to terminate the reaction; after methanol is cleaned, nitrogen is loaded for blow-drying, and after the reaction with acetic anhydride and dichloromethane in sequence, a dichloromethane layer is taken for machine detection. The polysaccharide derivative is loaded on a GC-MS instrument for analysis, the sample loading amount is 1 mu L, the carrier gas is helium, the initial 2min temperature is controlled at 140 ℃, then the temperature is gradually increased to 230 ℃ in a proportion of 3 ℃/min, and the mass scanning range of the mass spectrum is 30-600m/z.
Methylation analysis results: by analyzing the mass spectrum result after the methylation of the russula polysaccharide, comparing with the glycosidic bond fragments of a Complex Carbohydrate Research Center (CCRC) database, 9 glycosidic fragments are found in the russula polysaccharide, wherein the 4 glycosidic fragments with higher content, namely 6-Gal (p), t-Fuc (p), 2,6-Gal (p), t-Gal, have relative molar ratios of 74.48%,7.113%,6.056% and 4.6%, respectively. The results are shown in Table 1. The results indicate that branches may be present in the polysaccharide.
TABLE 1 methylation analysis of RGP1
TABLE 2 RGP1 1 H and 13 chemical shift of C
NMR analysis: preparing 40mg/mL of Rumex Gracilis polysaccharide solution, loading on a mass spectrometer of 500MHz at room temperature, and detecting the polysaccharide 1 H、 13 A C one-dimensional spectrum and a COSY, HSQC, HMBC, NOESY two-dimensional spectrum.
Results of NMR analysis: determining the chemical shift results for each residue in combination with literature results and NMR results, see table 2; the one-dimensional hydrogen spectrum signal of the russula polysaccharide is mainly concentrated between 3.0 and 5.5ppm, and the one-dimensional carbon spectrum signal is mainly concentrated between 55 and 110 ppm. As shown in Table 2 and the results in FIGS. 7-8, the chemical shift results of the respective residues are shown, the signal peaks of → 6) - α -D-Galp- (1- & gt, 6) - α -D-OMe-Galp- (1- & gt, the signal peaks of anomeric H and anomeric C are concentrated at 4.92ppm and 97.89/97.73ppm, the signal peaks of the terminal sugars α -L-Fucp- (1- & gt, the signal peaks of anomeric H and anomeric C are concentrated at 5.01ppm and 101.39ppm, and the signal peaks of the branched sugars → 2, 6) - α -D-Galp- (1- & gt, the signal peaks of anomeric H and anomeric C are concentrated at 4.99ppm and 98.35ppm; the correlation signal between adjacent C-H or H-H can be reflected according to two-dimensional signal spectra COSY and HSQC in the molecule, the result is shown in figures 9-10, NOESY and HMBC can react with the correlation signal between molecules, wherein significant signals of 3.37-78.81ppm and 3.5-56.08ppm exist, the polysaccharide structure has obvious H (OMe) -BC3 and C (OMe) -BH3 connection, namely, the substitution site of methoxy is the third position of 1, 6-galactose.
Example 3
To further verify the hematopoietic activity of russula polysaccharide (RGP 1), the following experiments were performed.
(1) Test materials: the purified russula polysaccharide, physiological saline, cyclophosphamide, recombinant human granulocyte colony-stimulating factor (rhG-CSF), balb/c mice of example 1;
(2) Test method and results
Mice were modeled and dosed: 90 Balb/c mice were randomly divided into 6 groups (n=15) which were a blank control group, a model group, a positive control group, a low/high dose russula polysaccharide dosing group, and a russula polysaccharide blank control dosing group. During modeling, 100mg/kg cyclophosphamide is injected into the abdominal cavity of a model group, a positive control group and a low/high-dose russula polysaccharide administration group mice for three consecutive days, and the other two groups of mice are given an equivalent amount of physiological saline; after modeling, 6 weeks of administration treatment was performed, wherein mice in the blank control group and the model group were perfused with physiological saline, mice in the positive control group were intraperitoneally injected with 30 μg/kg of rhG-CSF, and the low/high dose of russula polysaccharide administration group and russula polysaccharide blank control administration group were administered with 50, 100mg/kg of RGP1, respectively.
Sample collection: after the administration is completed, the mice are subjected to continuous water treatment during 8 hours of grain interruption, then peripheral blood of the mice is collected by adopting a tail vein blood sampling method, one part of the peripheral blood is placed in an anticoagulation tube for flow cytometry analysis, and after the other part of the peripheral blood is settled for 0.5 hour at room temperature, the other part of the peripheral blood is centrifuged at 3000rpm for 5 minutes to collect serum; next, taking the femur and tibia of the mouse in a sterile workbench, collecting bone marrow cells in one part of femur and tibia for flow cytometry analysis, fixing the other part of femur in 4% paraformaldehyde, and waiting for histopathological analysis; in addition, the spleens of mice were frozen in a-80℃refrigerator for use.
Flow cytometry analysis: collecting sterile bone marrow cells from femur and tibia using RPMI-1640 medium, and filtering through 70 μm filter to remove impurities such as broken bone; subsequently, erythrocytes in bone marrow cells and anticoagulated peripheral blood cells are removed using an erythrocyte lysis buffer; the number of living cells was analyzed using trypan blue staining, and the cell concentration was adjusted to 10 according to the result 6 100. Mu.L. According to the surface markers of stem and progenitor cells in bone marrow, the cell population is defined as follows: LSK (Lin) - Sca-1 + c-Kit + )、LK(Lin - Sca-1 - c-Kit + ) And LT-HSC (CD 48) - CD150 + LSK); lymphocytes in bone marrow are defined as follows: natural Killer (NK) cells (CD 3) - CD49b + ) B cells (CD 45) + CD19 + ). Peripheral blood T cell populations are defined as follows: activated T cells (CD 3 e) + CD28 + ) T helper cells (Th cells) (CD 3) + CD4 + ). The staining protocol used in this study was consistent with the antibody instructions. Immediately after the staining was completed, the cell was examined for cell clusters by flow cytometry in a dark environment.
Flow cytometry analysis results:
HSCs can differentiate into a variety of lymphocytes, providing the necessary hematopoietic cells for the organism. In hematopoietic dysfunctionIn the case of the hindered mice, rhG-CSF and RGP1 significantly increased lineage cells (Lin - Cell), lin - Sca-1 + c-kit + Cells (LSK cells), lin - Sca-1 - c-kit + Cells (LK cells) and Lin - Sca-1 + c-kit + CD48 - CD150 + Proportion of cells (LT-HSC). For lymphocytes, RGP1 significantly increases CD3e in peripheral blood + CD28 + Number of cells (activated T cells) and CD3 + CD4 + The number of cells (Th cells) without affecting CD3 in the bone marrow of hematopoietic dysfunction mice - CD49b + Cells (NK cells) and CD45 + CD19 + Expression of cells (B cells). The above results initially demonstrate that RGP1 is produced by the modulation of CD4 + The level of T cells relieves hematopoietic injury in mice. The results are shown in FIG. 13.
Histopathological observation: the fixed femur was decalcified using EDTA decalcification solution at room temperature until the bone softened. Decalcified femur was transferred to ethanol and xylene with stepwise increasing concentrations for dehydration treatment, then embedded in paraffin, and cut into 4 μm thick sections. Dewaxed sections were stained with hematoxylin and eosin and observed for pathological changes of tissue using an optical microscope.
Histopathological observations: according to the observation of the dyeing result, the bone marrow cells in the femur of the mice in the blank control group and the RGP1 blank administration group are closely and orderly arranged and have uniform density; modeling of cyclophosphamide reduces the number and cell density of bone marrow cells and creates a large number of vacuoles in the bone marrow cavity. This bone marrow damage is alleviated by RGP1 administration. The results are shown in FIG. 14.
Proteomic analysis: spleens of mice in a blank control group, a model group and a high-dose RGP1 administration group were taken to prepare tissue homogenates, and protein concentrations in each group of samples were detected by a BCA kit. Next, proteins were precipitated using acetone, and then the samples were subjected to reduction and alkylation treatments with TCEP and Iodoacetamide (IAA). After the proteins were trypsinized, peptide samples were collected and then separated by a nano ultra high performance liquid chromatography (UPLC) system equipped with a reverse phase chromatography column and then detected by Q-Exactive Orbitrap mass spectrometry. Raw data was analyzed using MaxQuant (2.0.1.0) and UNIPORT databases. When the protein expression or peptide expression fold difference between the two groups was not less than 2, the protein was defined as a significantly changed protein and was used for heat map and cluster analysis. Enrichment of protein interaction networks, gene Ontology (GO) and kyoto genes and genome encyclopedia (KEGG) was analyzed by the sting database.
Proteomic analysis results: RGP1 increased the level of 16 proteins in the mouse spleen compared to hematopoietic dysfunction mice, and the interactions between these proteins were analyzed based on the STRING database. According to GO analysis, signaling between 16 proteins involves T cell activation, CD4 receptor binding and hematopoietic regulation; according to KEGG analysis, these significantly altered proteins are mainly enriched in leukocyte activity and hematopoietic cell lineages. Among them, rac2, a hematopoietic specific gene, has been reported to be capable of affecting T cell activation and hematopoietic lineage cell activity; hcls1 are present in hematopoietic tissues or cells and are involved in T lymphocyte-associated signaling; IL-16 can bind to its receptor CD4, promoting secretion of downstream cytokines; annexin A1 plays a critical role in maintaining homeostasis of the immune system and regulating proliferation and differentiation of T cells. Thus, by proteomic analysis, RGP1 was found to be capable of modulating CD4 + The activity of T cells improves hematopoietic function. The results are shown in FIG. 15.
And (3) biochemical index detection: the secretion of CD 4-associated cytokines, and the level of hematopoietic regulatory factors in the serum and spleen of mice were analyzed using ELISA kits, and the specific methods of operation were consistent with the specification.
Biochemical index detection results: CD4 + The modeling of cyclophosphamide reduces IL-2, IL-3, IL-5, IL-6 levels in mice serum and spleen, increases expression of hematopoietic inhibitors TNF-alpha and TGF-beta, and the abnormal cytokine levels are reversed by RGP1 administration, as compared to the placebo mice, as shown in FIG. 16, which is the polysaccharide RGP1 of Myricus ruber on hematopoietic functionEffects of spleen and serum IL-2 levels in the mice with disorders; FIG. 17 is the effect of russula gray polysaccharide RGP1 on spleen and serum IL-3 levels in hematopoietic dysfunction mice; FIG. 18 is the effect of russula gray polysaccharide RGP1 on spleen and serum IL-5 levels in hematopoietic dysfunction mice; FIG. 19 is the effect of russula gray polysaccharide RGP1 on spleen and serum IL-6 levels in hematopoietic dysfunction mice; FIG. 20 is the effect of russula gray polysaccharide RGP1 on spleen and serum TNF- α levels in hematopoietic dysfunction mice; FIG. 21 is the effect of russula greysari polysaccharide RGP1 on spleen and serum TGF-beta levels in hematopoietic dysfunction mice.
Thus, as can be seen from FIGS. 16-21, the results of ELISA confirmed that RGP1 was able to regulate CD4 + Secretion of cytokines in T cells improves hematopoietic damage in hematopoietic dysfunction mice.
The technical scheme of the invention is not limited to the specific embodiment, and all technical modifications made according to the technical scheme of the invention fall within the protection scope of the invention.
Claims (4)
1. The russula cinerea polysaccharide with hematopoiesis promoting effect is characterized in that: the molecular weight of the strain is 31.4kDa, and the molecular weight of the strain is mainly determined by the molar mass ratio of 25.77:8.37:2.51:1.52: galactose, methoxy galactose, glucose, mannose, fucose of 1.41.
2. A method for preparing the russula polysaccharide with hematopoiesis promoting effect as claimed in claim 1, which comprises the following steps:
s1, taking and drying the russula vinosa fruiting bodies, and crushing the fruiting bodies by using a crusher;
s2, according to the mass of the russula vinosa fruiting bodies: deionized water volume = 1: mixing at 30-50 ratio, leaching with hot water at 60-90deg.C for 2-4 hr; repeating the extraction twice, combining the two extracting solutions, and concentrating the extracting solution by rotary evaporation;
s3, removing protein components in the extracting solution by using a Sevag method, discarding organic reagents, and performing rotary evaporation and dialysis on the upper extracting solution;
s4, adding an absolute ethanol solution into the dialyzed liquid to ensure that the final concentration of ethanol is 80%, fully and uniformly mixing, standing for 12 hours at 4 ℃, centrifuging at 6000rpm for 20 minutes after standing, collecting precipitate, drying the ethanol, and freeze-drying to obtain the russula vinosa Linne crude polysaccharide;
s5, dissolving the russula botrytis cinerea crude polysaccharide by using ultrapure water, wherein the concentration of the prepared polysaccharide solution is 0.05-0.1g/mL, passing through a 0.45um filter membrane after the polysaccharide solution is fully dissolved, loading the solution on a DEAE Sepharose FF ion exchange column, eluting mobile phases respectively into 0, 0.1 and 0.3M NaCl solutions, sequentially named RGP-A, RGP-B, RGP-C as polysaccharide components eluted by the mobile phases, and collecting 7mL of liquid per tube at the flow rate of 1-2 mL/min; measuring polysaccharide content in the collected liquid by adopting a phenol-sulfuric acid method, drawing an elution curve, collecting a polysaccharide tube with higher concentration, performing rotary evaporation, dialysis and freeze-drying;
s6, separating and purifying by using a DEAE chromatographic column to finally obtain two main polysaccharide components, namely RGP-A, RGP-B; using K562 cells, combining with an XTT method, an apoptosis detection method, a benzidine staining method and a western blot method to analyze and screen RGP-A with better in-vitro hematopoietic promotion activity to be applied to subsequent purification;
s7, sequentially loading RGP-A on HiPrepTM 26/60SephacrylTM S-400 and Ezload26/60Chromdex 200pg gel columns, eluting the mobile phase with 0.15M NaCl, and collecting 2-4mL of liquid in each tube at a flow rate of 1 mL/min; and (3) measuring the polysaccharide content in the collected liquid by adopting a phenol-sulfuric acid method, drawing an elution curve, collecting a polysaccharide tube with higher concentration, performing rotary evaporation, dialysis and freeze-drying to obtain the purified polysaccharide of the russula cinerea with hematopoiesis promoting effect.
3. Use of the russula polysaccharide with hematopoiesis promoting effect according to claim 1 for preparing a medicine for promoting hematopoiesis.
4. The use of russula vinosa polysaccharides with hematopoiesis promoting effect according to claim 3 for preparing a medicament for promoting hematopoiesis, characterized in that: the russula vinosa polysaccharide with hematopoiesis promoting effect is used for preparing medicines for relieving hematopoietic dysfunction caused by chemotherapy.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19990062375A (en) * | 1997-11-20 | 1999-07-26 | 김상희 | Compositions for Promoting Proliferation of Hematopoietic Stem Cells and Platelet Progenitor Cells Purified from Deer Antlers and Purification Methods Thereof |
| CN107050044A (en) * | 2017-07-04 | 2017-08-18 | 吉林大学 | Medical application of the grey meat Russula polysaccharide in systemic lupus erythematosus |
| CN111778163A (en) * | 2020-05-15 | 2020-10-16 | 广东省微生物研究所（广东省微生物分析检测中心） | New russula griseofulva endophytic fungus and application of extract thereof in bacteriostasis |
| CN112569336A (en) * | 2020-12-16 | 2021-03-30 | 广东省微生物研究所（广东省微生物分析检测中心） | Application of grifola frondosa glycoprotein in preparation of drugs or health foods for enhancing body immunity and preventing and/or treating tumor diseases |
-
2023
- 2023-01-17 CN CN202310055533.9A patent/CN116693707B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR19990062375A (en) * | 1997-11-20 | 1999-07-26 | 김상희 | Compositions for Promoting Proliferation of Hematopoietic Stem Cells and Platelet Progenitor Cells Purified from Deer Antlers and Purification Methods Thereof |
| CN107050044A (en) * | 2017-07-04 | 2017-08-18 | 吉林大学 | Medical application of the grey meat Russula polysaccharide in systemic lupus erythematosus |
| CN111778163A (en) * | 2020-05-15 | 2020-10-16 | 广东省微生物研究所（广东省微生物分析检测中心） | New russula griseofulva endophytic fungus and application of extract thereof in bacteriostasis |
| CN112569336A (en) * | 2020-12-16 | 2021-03-30 | 广东省微生物研究所（广东省微生物分析检测中心） | Application of grifola frondosa glycoprotein in preparation of drugs or health foods for enhancing body immunity and preventing and/or treating tumor diseases |
Non-Patent Citations (5)
| Title |
|---|
| LIU, X 等: "Structural characterization of Russula griseocarnosa polysaccharide and its improvement on hematopoietic function", INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, vol. 263, 30 April 2024 (2024-04-30), pages 1 - 10 * |
| LIU, Y 等: "Purification, characterization and anti-tumor activities of polysaccharides extracted from wild Russula griseocarnosa", INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, vol. 109, 30 April 2018 (2018-04-30), pages 1054 - 1060 * |
| QIAN CHEN 等: "Immune-enhancing effects of a polysaccharide PRG1-1 from <italic>Russula griseocarnosa</italic> on RAW264.7 macrophage cells via the MAPK and NF-kappaB signalling pathways.", FOOD AND AGRICULTURAL IMMUNOLOGY, vol. 29, no. 01, 31 December 2018 (2018-12-31), pages 833 - 844 * |
| 刘鑫 等: "灰肉红菇多糖的结构表征及其基于免疫调节的促造血功能研究", 中国优秀博士论文全文数据库医药卫生科技, 15 December 2023 (2023-12-15), pages 1 - 206 * |
| 陈志伟;祝彼得;许惠玉;: "肉苁蓉多糖对骨髓抑制性贫血小鼠造血调控的实验研究", 中华中医药学刊, no. 07, 10 July 2007 (2007-07-10), pages 1473 - 1474 * |
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