NL2032676B1 - Neutral oligosaccharide cordyceps militaris for regulating lung lymphocyte differentiation, preparation method and applications thereof - Google Patents
Neutral oligosaccharide cordyceps militaris for regulating lung lymphocyte differentiation, preparation method and applications thereof Download PDFInfo
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- cordyceps militaris
- neutral oligosaccharide
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- 241001264174 Cordyceps militaris Species 0.000 title claims abstract description 81
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7004—Monosaccharides having only carbon, hydrogen and oxygen atoms
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A61K31/00—Medicinal preparations containing organic active ingredients
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- A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/062—Ascomycota
- A61K36/066—Clavicipitaceae
- A61K36/068—Cordyceps
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- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
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- C07H3/02—Monosaccharides
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Abstract
Disclosed are a neutral oligosaccharide of Cordyceps militaris for regulating lung lymphocyte differentiation, a method for preparing the neutral oligosaccharide and applications thereof, belonging to the field of natural product development and utilization. The neutral oligosaccharide of Cordyceps militaris includes dextrose, mannose, galactose, xylose, arabinose, rhamnose, ribose and rock in a mass ratio of 269.45 : 72.01 : 38.47 : 9.99 : 6.27 : 1.42 : 1.19 : 1. Neutral oligosaccharide of Cordyceps militaris can lower lung index, depress serum immune globulin E levels, increase interferon gamma levels, inhibit the expression of lL-4, lL-5, lL-13 and lL-17A inflammatory factors, promote Nrf-2 signalling pathway transduction and inhibit NF-KB signalling pathway transduction in mice with allergic asthma, thereby regulating the level of Th1/Th2 differentiation in mouse lymphocytes and alleviating the symptoms of lung inflammation.
Description
NEUTRAL OLIGOSACCHARIDE CORDYCEPS MILITARIS FOR REGULATING LUNG
LYMPHOCYTE DIFFERENTIATION, PREPARATION METHOD AND APPLICATIONS
THEREOF
The disclosure relates to the field of development and utilization of natural products, and in particular to a neutral oligosaccharide of Cordyceps militaris for regulating lung lymphocyte differentiation, preparation method and applications thereof.
Lymphocyte differentiation in lungs leads to a variety of lung diseases, such as allergic asthma, chronic obstructive pulmonary disease and pneumonia, all of which are caused by inflammation in lungs, and the severity of inflammation is largely influenced by lymphocyte differentiation in the lungs. Allergic asthma alone is an inflammatory lung disease that affects 300 million people worldwide and the number of patients is increasing at a rapid rate of 20 percent (%) - 25% every 10 years. Inflammatory lung diseases result not only in breathing difficulties, loss of appetite and weight, depression and anxiety in patients, but also place a heavy burden on governments and healthcare authorities in all countries.
Cordyceps militaris, also known as northern Chinese caterpillar fungus, belongs to the phylum ascomycota, order hypocreales, family clavicipitaceae; it is a medicinal fungus with high dietary value and biological activities such as antitumour, antibacterial and antioxidant; however, the role of its oligosaccharide in regulating lymphocyte differentiation in the lung has not been reported yet.
The present disclosure provides a neutral oligosaccharide of Cordyceps militaris for regulating lung lymphocyte differentiation, a preparation method and applications thereof so as to solve the problems existing in the prior art. The neutral oligosaccharide of Cordyceps militaris can lower the lung index and regulate the differentiation of lymphocytes in mice with allergic asthma, thus alleviating the symptoms of the case in mice.
To achieve the above objectives, the present disclosure provides the following technical schemes: a neutral oligosaccharide of Cordyceps militaris for regulating lung lymphocyte differentiation, comprising dextrose, mannose, galactose, xylose, arabinose, rhamnose, ribose and fucose in a mass ratio of 269.45 : 72.01: 38.47 : 9.99 : 6.27 : 1.42: 1.19:1.
Optionally, the neutral oligosaccharide of Cordyceps militaris is a beta (B)-furanose.
The present disclosure also provides a preparation method of the neutral oligosaccharide of
Cordyceps militaris for regulating lung lymphocyte differentiation, comprising the following steps: (1) extraction of Cordyceps militaris polysaccharide pulverizing dried Cordyceps militaris into powder, adding the pulverized Cordyceps militaris powder into ethanol solution, standing the solution for solid-liquid separation after ultrasonic treatment, performing ultrasonic treatment on the remaining precipitate in ethanol solution, standing again for separation, taking out the precipitate, and drying the precipitate to obtain
Cordyceps militaris powder; adding Cordyceps militaris powder into water, extracting by wall-breaking under high temperature, centrifuging the extracted powder to obtain a supernatant, adding absolute ethanol into the supernatant; centrifuging the supernatant added with absolute ethanol after standing to obtain a supernatant, concentrating the centrifuged supernatant obtain a supernatant, and freeze-drying the concentrated supernatant to obtain a crude polysaccharide of Cordyceps militaris; the obtained crude polysaccharide of Cordyceps militaris is de-proteinized by sevag method, dialyzed to remove micromolecule compounds and then freeze-dried to obtain polysaccharide of Cordyceps militaris; (2) column chromatography dissolving Cordyceps militaris polysaccharide, purifying the dissolved Cordyceps militaris polysaccharide solution with cellulose anion exchange chromatography column, dialyzing and concentrating the purified solution, and freeze-drying the concentrated solution to obtain neutral oligosaccharide of Cordyceps militaris.
Optionally, the ethanol solution is in a volume concentration of 80%.
Optionally, the purifying is carried out in a gradient elution using sodium chloride solution.
The present disclosure also provides an application of the neutral oligosaccharide of
Cordyceps militaris for regulating lung lymphocyte differentiation in preparing health products or medicines for regulating lung lymphocyte differentiation.
Optionally, the lymphocytes include lymphocytes of T helper cell type 1 (Th1) and T helper cell type 2 (Th2).
The present disclosure also provides an application of the neutral oligosaccharide of
Cordyceps militaris for regulating lung lymphocyte differentiation in preparing health products or medicines for treating allergic asthma.
The present disclosure also provides a health product or medicine containing the neutral oligosaccharide of Cordyceps militaris for regulating lung lymphocyte differentiation.
The present disclosure discloses the following technical effects: according to the present disclosure, the crude polysaccharide of Cordyceps militaris is extracted by ethanol ultrasonic extraction and the method of mechanically crushing under high-
temperature, and free proteins and micromolecule compounds are removed to obtain
Cordyceps militaris polysaccharide; the obtained polysaccharide has a sugar content of 66.49 + 0.47%, an uronic acid content of 12.94 + 1.55%, and a protein content of 26.54 + 0.71%; polysaccharide of Cordyceps militaris is purified by a DEAES2 (Diethylaminoethyl) cellulose column chromatography to obtain the neutral oligosaccharide of Cordyceps militaris, the obtained neutral oligosaccharide is a B-furanose with a molecular weight of 15.94 Kilodalton (KDa), also with a monosaccharide composition in a ratio of dextrose : mannose : galactose : xylose : arabinose : rhamnose : ribose : fucose = 269.45: 72.01: 38.47: 9.99: 6.27 : 1.42: 1.19 : 1; in vivo experiments in mice confirm that neutral oligosaccharide of Cordyceps militaris can lower lung index, depress serum immune globulin E (IgE) levels, increase interferon gamma (INF-y) levels, inhibit the expression of IL-4, IL-5, IL-13 and IL-17A inflammatory factors, promote nuclear factor erythroid 2-related factor 2 (Nrf-2) signalling pathway transduction and inhibit Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) signalling pathway transduction in mice with allergic asthma, thereby regulating the level of Th1/Th2 differentiation in mouse lymphocytes and alleviating the symptoms of lung inflammation.
For a clearer illustration of the technical schemes in the embodiments of the present disclosure or in the prior art, a brief description of the accompanying figures to be used in the embodiments will be given below. It is obvious that the accompanying figures in the following description are only some embodiments of the present invention, and that other accompanying figures may be obtained on the basis of these figures without any creative effort on the part of a person of ordinary skill in the art.
Fig. 1 shows a standard curve of dextrose content.
Fig. 2 illustrates a standard curve of protein content.
Fig. 3 shows a standard curve of uronic acid content.
Fig. 4 illustrates a standard curve of molecular weight of standard dextran.
Fig. 5 is a high performance gel permeation chromatogram of neutral oligosaccharide of
Cordyceps militaris.
Fig. 6 illustrates a HPLC chart of monosaccharide composition of neutral oligosaccharide of
Cordyceps militaris.
Fig. 7 shows an infrared spectrum of Cordyceps militaris polysaccharide.
Fig. 8 shows the effect of cytidine monophosphate (CMP) on serum immune globulin E (IgE) level in ovalbumin-induced (OVA-induced) allergic asthma mice. Note: compared with the Control group, **p < 0.01, ***p < 0.001; compared with OVA group, #p < 0.05, ##p < 0.01, ##4p < 0.001.
Fig. 9 shows the effect of CMP on interferon gamma (INF-y) level in lung tissue of OVA- induced mice with allergic asthma. Note: compared with the Control group, ***p < 0.001; compared with OVA group, ##p < 0.01.
Fig. 10 shows the effect of CMP on interleukin-4 (IL-4) level in lung tissue of OVA-induced mice with allergic asthma. Note: compared with the Control group, **p < 0.01, ***p < 0.001; compared with OVA group, ###p < 0.001.
Fig. 11 shows the effect of CMP on IL-5 level in lung tissue of OVA-induced mice with allergic asthma. Note: compared with the Control group, **p < 0.01, ***p < 0.001; compared with OVA group, ###p < 0.001.
Fig. 12 shows the effect of CMP on IL-13 level in lung tissue of OVA-induced mice with allergic asthma. Note: compared with the Control group, ***p < 0.001; compared with OVA group, #p < 0.05, ##p < 0.001.
Fig. 13 shows the effect of CMP on the level of IL-17A in lung tissue of OVA-induced mice with allergic asthma. Note: compared with the Control group, ***P < 0.001; compared with
OVA group, ###p < 0.001.
Fig. 14 shows the effect of CMP on lung histopathology of OVA-induced mice with allergic asthma; in which, A is the Control group, B is Group OVA, C is OVA + dexamethasone (OVA + DEX) group, D is OVA + CMP 200 group; E is OVA + CMP 100 group; F is
OVA + CMP 50 group.
Fig. 15 shows the H-Score levels of nuclear factor erythroid 2-related factor 2 (Nrf-2) and P-
P65 protein in lung tissue. Note: compared with the Control group, *p < 0.05, **p < 0.01; compared with OVA group, #p < 0.05, ##p < 0.01.
Now, various exemplary embodiments of the present application will be described in detail.
This detailed description should not be taken as a limitation of the present application, but should be understood as a more detailed description of some aspects, characteristics and embodiments of the present application.
It should be understood that the terms mentioned in the present application are only used to describe specific embodiments, and are not used to limit the present application. In addition, for the numerical range in the present application, it should be understood that each intermediate value between the upper limit and the lower limit of the range is also specifically disclosed.
Every smaller range between any stated value or the intermediate value within the stated range and any other stated value or the intermediate value within the stated range is also included in the present application. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.
Unless otherwise stated, all technical and scientific terms used herein have the same meanings commonly understood by those of ordinary skill in the field to which this application relates. Although the present application only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and/or materials related to the 5 documents. In case of conflict with any incorporated documents, the contents of this specification shall prevail.
Without departing from the scope or spirit of the present application, it is obvious to those skilled in the art that many modifications and changes can be made to the specific embodiments of the present specification. Other embodiments obtained from the description of the present application will be obvious to the skilled person. The description and embodiment of that application are only exemplary.
As used in this paper, the terms "comprising", "including", "having" and "containing" are all open terms, meaning including but not limited to.
Embodiment 1 Preparation and structure analysis of Cordyceps militaris polysaccharide 1.1 Extraction of Cordyceps militaris polysaccharide preparing 200 grams (g) of dried Cordyceps militaris, crushing the dried Cordyceps militaris, adding 80 volume percent (vol.%) ethanol solution at a ratio of material to liquid of 1: 20 (g : milliliter (mL)) into the crushed Cordyceps militaris, followed by ultrasonic at 500 Watts (W) and 55 degree Celsius (°C) for 20 minutes (min); pouring out a supernatant after standing the ultrasonicated Cordyceps militaris for 2 hours (h); adding 80 vol.% ethanol solution to a remaining precipitate at a ratio of material to liquid of 1 : 10 (g : mL), followed by ultrasonic at 500 W and 55°C for 20 min; pouring out a supernatant after standing the ultrasonicated
Cordyceps militaris for 2 h; collecting that remaining precipitate, heating and drying the precipitate at 50°C to obtain Cordyceps militaris powder; weighing Cordyceps militaris powder, adding its volume of hot water at 80°C, pouring the powder with water together into a wall-breaking machine, boiling the powder and water for wall- breaking extraction for 8 min, centrifuging the boiled powder and water to separate supernatant, followed by adding 3 times of absolute ethanol and standing for 12 h; centrifuging again to collect supernatant, which is concentrated to one third of the original liquid volume, centrifuging the concentrated supernatant at 7,000 revolutions per minute (rpm) for 10 min, collecting a supernatant, and freeze-drying to obtain crude polysaccharide of Cordyceps militaris; de-proteinizing the obtained crude polysaccharide of Cordyceps militaris by sevag method, dialyzing the de-proteinized crude polysaccharide to remove micromolecule compounds, and then freeze-drying the dialyzed crude polysaccharide to obtain Cordyceps militaris polysaccharide, where the obtained polysaccharide has a sugar content of 66.49 + 0.47%, an uronic acid content of 12.94 + 1.55%, and a protein content of 26.54 £ 0.71%; 1.2 DEAE-52 cellulose column chromatography dissolving the obtained Cordyceps militaris polysaccharide in distilled water and purifying the dissolved Cordyceps militaris polysaccharide by DEAE 52 (Diethylaminoethyl) cellulose anion exchange chromatography column (4.5 centimeters (cm) x 40 cm}; carrying out gradient elution with 0, 0.1, 0.2 mol/l NaCl solution; collecting one tube of eluent every 3 mL, tracing and detecting the polysaccharide content by phenol-concentrated sulfuric acid method, collecting the polysaccharide components eluted by distilled water, dialyzing and concentrating the collected polysaccharide components, and freeze-drying the concentrated polysaccharide components to obtain neutral oligosaccharide of Cordyceps militaris; 1.3 Determination of molecular weight of Cordyceps militaris polysaccharide determining the molecular weight of neutral oligosaccharide of Cordyceps militaris by high performance gel permeation chromatography, and determining the molecular weight of the sample according to the retention time of the sample, where the smaller the retention time, the greater the molecular weight, and the chromatographic conditions are RID-10A parallax refractive detector, TSK-gel G-3000PWXL column (7.8x300 nanometer (nm)), mobile phase of distilled water, flow rate of 0.6 mL/min, column temperature of 35°C, and sample volume of 10 microliters (ul);
FIGS. 1-3 show the content standard curves of dextrose, protein and uronic acid, FIG. 4 shows the molecular weight standard curve of standard dextran, and a molecular weight standard curve equation y = -0.158x + 7.7965, R? = 0.9949 is obtained; FIG. 5 shows the gel permeation chromatography structure, where a molecular weight of neutral oligosaccharide of
Cordyceps militaris is calculated to be 15.94 Kilodaltons (KDa) according to the retention duration and standard curve; 1.4 Analysis of monosaccharide composition of Cordyceps militaris polysaccharide taking a clean chromatographic bottle, adding 5 milligrams (mg) of neutral oligosaccharide of Cordyceps militaris into a prepared trifluoroacetic (TFA) acid solution, and heating the polysaccharide-added solution at 121°C for 2 h; blow-drying the solution with nitrogen, washing the blow-dried solution with methanol, followed by blow-drying again; repeating the washing and blow-drying for 3 times; dissolving the dried sample in sterile water, and transferring the dissolved sample to chromatographic bottle for testing, where the conditions of mobile phase are as follows: phase A: ddH2O; phase B: 200 Millimolars (mM) NaOH/500 mM NaAC; flow rate: 0.5 mL/min; according to the results shown in FIG 8, the neutral oligosaccharide of Cordyceps militaris is consisted of dextrose : mannose : galactose : xylose : arabinose : rhamnose : ribose : fucose in a ratio of 269.45 : 72.01 : 38.47 : 9.99 : 6.27 : 1.42: 1.19: 1; 1.5 Infrared spectrum analysis of Cordyceps militaris polysaccharide weighing 1.5 mg neutral oligosaccharide of Cordyceps militaris and 200 mg chromatographically pure potassium bromide (KBr), mixing the polysaccharide and KBr fully and grinding the mixed polysaccharide and KBr with agate mortar, tabletting the ground mixture, and then tabletting 200 mg KBr separately as background; scanning and analysing the infrared spectrum of tablet by infrared spectrometer in the range of 4,000 - 500 cm, and the results as shown in FIG. 7 indicate that the neutral oligosaccharide of Cordyceps militaris is beta (B)- furanose.
Embodiment 2 Study on the mechanism of Cordyceps militaris polysaccharide improving lymphocyte differentiation activity in asthmatic mice 2.1 Design and treatment of animal experiments 2.1.1 Animal grouping randomly dividing 90 Balb/c mice into 6 groups, namely, blank control group (Control group), model group (OVA group), dexamethasone group (OVA + DEX group), Cordyceps militaris neutral oligosaccharide high-dose group (OVA + cytidine monophosphate (CMP) 200 group, Cordyceps militaris neutral oligosaccharide medium-dose group (OVA + CMP 100 group), Cordyceps militaris neutral oligosaccharide low-dose group (OVA + CMP 50 group), 15 in each group; 2.1.2 Establishment and administration of animal models injecting 0.2 mL of sensitizing solution intraperitoneally on days 1, 8 and 15 of the start of the moulding for each group of mice except the Control group, and injecting an equal volume of saline intraperitoneally for the mice of Control group; administering each group at a fixed time every day, where the 22 - 35" day is the challenge stage; calculating the dosage of OVA + CMP 200 group, OVA + CMP 100 group and OVA + CMP 50 group according to the difference of standard weight and drug resistance between human and animal to be 200 mg/kg (kilogram), 100 mg/kg and 50 mg/kg respectively, and that of OVA + DEX group is 2.5 mg/kg; providing the
Control group and the OVA group with corresponding volume of saline; performing nebulisation attack 1 h after the end of gavage in each group, with 30 min per attack and once a day, while nebulisation attack in the Control group is replaced by saline; 2.1.3 Animal handling and sample collection fasting the mice while providing water for 24 h after the final treatment, collecting blood from mice by removing eyeballs and centrifuging the blood twice at 4°C and 2,000 rpm for 10 min, separating the serum and storing the serum at -80°C in separate packages; opening the thoracic cavity of the mouse, taking the tissue of the lower lobe of the right lung and part of the duodenum and fixing the tissue in 4% paraformaldehyde solution; storing the rest of the lung tissue and the remaining parts of the duodenum, cecum and colon at -80°C for later use; 2.2 Mouse lung index recording the body weight of mice before dissection and the wet weight of lung tissue after dissection, calculating the lung index according to a formula as follows: lung index = lung weight/body weight* 100%, where the results are shown in Table 1, and it can be seen from the results that as comparing to the Control group, lung indexes in other five groups show no increase, with significant differences exist among OVA group, OVA + CMP 200 group, OVA +
CMP 100 group and OVA + CMP 50 group (p < 0.05), and as comparing to the OVA group, the lung indexes in other five groups are all decreased, among which OVA + DEX group, OVA +
CMP 200 group and OVA + CMP 50 group show significant differences (p < 0.05), and the values of OVA + DEX group and OVA + CMP 200 group are the closest to that of Control group.
Table 1 Comparison of lung index of mice in each group ~~ Control 074x010
OVA 1.42 + 0.20%
OVA + DEX 0.80 + 0.07#
OVA + CMP 200 0.8810.13°#
OVA + CMP 100 1.40 + 0.25“
OVA + CMP 50 1.24 + 0.23% note: compared with the Control group, *p < 0.05; compared with the OVA group, #p < 0.05 2.3 ELISA for measuring serum IgE and levels of IL-4, IL-5, IL-13, INF-y and IL-17A in lung tissue detecting the immune globulin E (IgE) content in serum and the contents of interleukin-4 (IL-4), IL-5, IL-13, interferon gamma (INF-y) and IL-17A in lung tissue by enzyme-linked immunosorbent assay (ELISA) strictly according to the instructions of ELISA kit, where the results are shown in FIGS. 8-13 and it can be seen from the figures that the content of INF-y in lung tissue (p < 0.001) in OVA-induced allergic asthma mice is significantly reduced; the content of INF-y is increased after administered with CMP, while that in the OVA + CMP 100 group and
OVA + CMP 50 group shows no significant difference, and the content in the OVA + CMP 200 group shows a significant difference compared with that in the OVA group (p < 0.01); contrary to
INF-y, the levels of IL-4, IL-5, IL-13 and IL-17A in lung tissue of OVA group are higher than those of Control group (p < 0.001); the levels of IL-4, IL-5, IL-13 and IL-17A in the lung tissues of OVA-induced allergic asthmatic mice are also significantly inhibited by 200 mg/mL CMP (p < 0.001), with the inhibitory effects on IL-4, IL-5 and IL-17A being better than those in the OVA +
DEX group, this indicates that neutral oligosaccharide of Cordyceps militaris can reverse the effect of OVA on cytokines related to mouse lung tissue; 2.4 Pathological observation of lung tissue fixing the lower lobe of the right lung of mice with 4% paraformaldehyde solution, embedding the fixed lower lobe in paraffin, and staining the embedded lower lobe with hematoxylin-eosin (HE) after routine sectioning, observing and photographing the pathological changes of four visual fields which are not repeated in each section under 200 times light microscope (FIG. 14), from which it can be seen that as comparing to the Control group, other groups show different degrees of structural abnormalities in lung tissues, among which OVA group and OVA + CMP 50 group show severe abnormalities, where some bronchial epithelial cells show irregularly arrangement and shedding off, and there is alveolar epithelial cells proliferation and thickening of alveolar diaphragm in addition to some alveoli atrophy, the tissue parenchyma is serious; in OVA + CMP 50 group, inflammatory cells are found in tracheal lumen; in OVA + CMP 50 group, alveolar epithelial cell necrosis and nuclear fragmentation are observed, and a large number of necrotic cell fragments can be seen; both the OVA + CMP 200 group and the OVA + CMP 100 group show moderate abnormalities in the lung tissue structure, with some thickening of the alveolar septa, mild widening of the alveolar walls and partially atrophy of alveolar, in addition to a reduced tissue parenchyma comparing to that in the OVA group; a small amount of diffuse infiltration of inflammatory cells is also seen in the tissue, and a small number of neutrophils are seen in the blood vessels. The bronchial epithelial cells are irregularly arranged and slightly detached. 2.5 Detection of inflammatory pathway-related protein expression in lung tissue by immunohistochemistry measuring the levels of Nrf-2 and P-P65 proteins in mouse lung tissue by immunohistochemistry, and analysing their histochemistry score (H-score), where it can be seen from the results as shown in FIG. 15 that there is no significant decrease in Nrf-2 protein levels in mice in the OVA group compared to the Control group, while mice treated in CMP 200 (p < 0.01), CMP 100 (p < 0.01), CMP 50 and DEX (p < 0.01) show increased levels comparing to that in the OVA group; the trend of P-P85 changes in contrast to that of Nrf-2, with mice induced by OVA exhibit a significant increased level of P-P65 (p < 0.01) compared to that in the Control group, whilst CMP 200 (p < 0.01), CMP 100 (p < 0.01) and DEX (p < 0.01) treatments all result in an inhibition of P-P65 levels; the above results suggest that the relief of OVA-induced asthma symptoms by CMP may be achieved by promoting the Nrf-2 signalling pathway and inhibiting the NF-kB signalling pathway, and that the effect of CMP treatment correlates with the concentration of CMP in a gradient, with the improvement effect of CMP 200 in allergic asthmatic mice being close to that of DEX.
The above-mentioned embodiments only describe the preferred mode of the application, but do not limit the scope of the application. On the premise of not departing from the design spirit of the application, all kinds of modifications and improvements made by ordinary technicians in the field to the technical scheme of the application shall fall within the scope of protection determined by the claims of the application.
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| CN112076207A (en) * | 2020-09-27 | 2020-12-15 | 延边大学 | High molecular weight cordyceps militaris polysaccharide, preparation method thereof and application of high molecular weight cordyceps militaris polysaccharide in preparation of anticomplement medicines |
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| ZHANG JIXIAN ET AL: "Advance in Cordyceps militaris (Linn) Link polysaccharides: Isolation, structure, and bioactivities: A review", INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, vol. 132, 1 July 2019 (2019-07-01), NL, pages 906 - 914, XP093004376, ISSN: 0141-8130, Retrieved from the Internet <URL:https://www.sciencedirect.com/science/article/pii/S014181301930995X/pdfft?md5=405fe8df3e0f449e0928c9ec5fb55a22&pid=1-s2.0-S014181301930995X-main.pdf> DOI: 10.1016/j.ijbiomac.2019.04.020 * |
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