CN113842450A - Application of ganoderma lucidum spore powder polysaccharide peptide in preparation of medicine for treating cancer-induced fatigue - Google Patents
Application of ganoderma lucidum spore powder polysaccharide peptide in preparation of medicine for treating cancer-induced fatigue Download PDFInfo
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- A61K36/074—Ganoderma
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- C—CHEMISTRY; METALLURGY
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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Abstract
The invention discloses an application of ganoderma lucidum spore powder polysaccharide peptide in preparing a medicament for treating cancer-induced fatigue. The polysaccharide peptide of the ganoderma lucidum spore powder is an effective site group extracted and enriched from ganoderma lucidum spores, and is a mixture, wherein the effective components with the highest content are polysaccharide and polypeptide, the polysaccharide content is more than 40%, and the polypeptide content is more than 20%. The test proves that the ganoderma lucidum spore powder polysaccharide peptide has obvious relieving effect on cancer-induced fatigue and has a drug development prospect.
Description
Technical Field
The invention belongs to the technology of medicine extraction and application, and particularly relates to application of ganoderma lucidum spore powder polysaccharide peptide in preparation of a medicine for treating cancer-induced fatigue.
Background
Cancer-related fatigue (CRF), also known as cancer-related fatigue, is one of the common symptoms of clinical malignancies, and the NCCN guidelines describe CRF as "an unsettling, persistent, subjective feeling of fatigue and failure of energy in the body, emotion and/or cognition, and interfering with daily life and function caused by tumor or anti-tumor therapy"; international disease classification standard 10 th edition (ICD 10) describes the symptoms of CRF as nonspecific weakness, general decline, lethargy, fatigue. It has persistent and non-universal characteristics, such as weakness, exercise intolerance, inattention, reduced motivation or interest; broadly, CRF is a multi-dimensional subjective experience of individual patients in terms of physiology, psychology, functionality and sociality.
In summary, CRF is a subjective feeling of fatigue, usually characterized by a decrease in physical strength and energy, due to a series of subjective feelings resulting from long-term stress and pain of patients caused by cancer and related treatments; unlike general fatigue, CRF has characteristics of high severity, disproportionate to activity or energy output, inability to be alleviated by sleep and rest, long duration, etc.; in clinic, there are various manifestations such as lack of energy, weakness, laziness, apathy, poor thought, hypomnesis, depression, etc.; CRF can seriously affect the physical, psychological and social functioning of the patient, as well as the quality of life.
Ganoderma lucidum (scientific name: Ganoderma lucidum Karst): also called Ruicao, SHENZHI, XIANCAO, Yao grass, caulis et folium Gaultheriae Yunnanensis, LINZHONGLING, JUNLINGZHI, WANNIAN mushroom, LINGCAO, CHIZHI, DANZHI, QINGZHEN, etc., is a fruiting body of Ganoderma lucidum belonging to Polyporaceae family; the shape of the mushroom cap is umbrella-shaped, kidney-shaped, semicircular or nearly circular; ganoderma lucidum is recorded in Ben Cao gang mu compiled by Li Shi Zhen of Ming Dynasty medical scientists; the ganoderma lucidum spores are extremely tiny ovoid germ cells, namely seeds of the ganoderma lucidum, which are ejected from ganoderma lucidum folds in the growth and maturation period of the ganoderma lucidum; each ganoderma lucidum spore is only 4-6 microns and is a living organism; the Ganoderma spore powder has all genetic materials of Ganoderma and health promotion effect; the medicinal value of the ganoderma lucidum spore is increasingly emphasized, and researches show that the ganoderma lucidum spore has the effects of enhancing the immunity of organisms, inhibiting tumors, protecting liver injury, protecting radiation and the like.
The ganoderma lucidum spore powder polysaccharide is a main active ingredient in ganoderma lucidum, has proved to have various biological activities and has wide application prospect in the field of medicine; at present, most of ganoderma lucidum spore polysaccharide extraction adopts a method of ultrasonic extraction, water extraction, concentration and drying, the content of polysaccharide is 4-8%, and the polysaccharide in ganoderma lucidum spores cannot be enriched; the application further improves the process for extracting the polysaccharide peptide from the ganoderma lucidum spore powder, and researches show that the polysaccharide peptide from the ganoderma lucidum spore powder also has obvious effect on the aspect of preparing the medicine for treating cancer-induced fatigue.
Disclosure of Invention
The purpose of the invention is as follows: aiming at the prior art, the invention provides the application of ganoderma lucidum spore powder polysaccharide peptide in preparing the medicine for treating cancer-induced fatigue.
The technical scheme is as follows: the invention discloses an application of ganoderma lucidum spore powder polysaccharide peptide in preparing a medicament for treating cancer-induced fatigue.
Wherein, the polysaccharide content in the ganoderma lucidum spore powder polysaccharide peptide is more than 40 percent, and the polypeptide content is more than 20 percent.
Further, the preparation method of the ganoderma lucidum spore powder polysaccharide peptide comprises the following steps:
(1) screening plump ganoderma lucidum spores for wall breaking;
(2) adding water into wall-broken Ganoderma spore powder, and making into strip granuleGranulating, drying at low temperature until the water content is less than 6%, and adding CO2Supercritical extraction of fat-soluble components;
(3) and adding water into the degreased wall-broken ganoderma lucidum spore particles, preserving heat at 90-100 ℃ for 1-3 hours, filtering, repeatedly treating filter residues for 1-3 times, combining filtrates, concentrating, adding 95% edible ethanol, stirring, standing overnight, taking precipitates, adding absolute ethyl alcohol, cleaning, and drying at low temperature to obtain the ganoderma lucidum spore powder.
In the step (1), the ganoderma lucidum spore powder raw material is preferably selected, so that the polysaccharide of the wall-broken ganoderma lucidum spore powder is not less than 1.0%, and the protein is not less than 1.0%.
In the step (2), the water amount is 1/5-1/4 of the weight of the wall-broken ganoderma lucidum spore powder.
Preferably, in the step (2), CO2The supercritical extraction conditions comprise that the extraction pressure is 25-28 MPa, the extraction temperature is 25-30 ℃, and the extraction time is 1-2 hours; most preferred conditions are: the extraction pressure is 25MPa, the extraction temperature is 30 ℃, and the extraction time is 2 hours.
In the step (3), adding 8-10 times of water by weight into the degreased wall-broken ganoderma lucidum spore particles, heating at 90-100 ℃, preserving heat for 1-3 hours, filtering, adding 6-8 times of water by weight into filter residues, heating at 90-100 ℃, preserving heat for 1-3 hours, filtering, and combining filtrates.
And (3) combining the filtrates, concentrating to a specific gravity of 1.15-1.23, adding 95% edible ethanol with 2-3 times of volume, stirring, standing overnight, and taking a precipitate.
In the step (3), the drying temperature is 30-40 ℃.
Has the advantages that: the polysaccharide peptide of the ganoderma lucidum spore powder is an effective site group extracted and enriched from ganoderma lucidum spores, and is a mixture, wherein the effective components with the highest content are polysaccharide and polypeptide; the test proves that the ganoderma lucidum spore powder polysaccharide peptide has obvious relieving effect on cancer-induced fatigue and has a drug development prospect.
Detailed Description
The invention is further illustrated by the following specific examples.
Example 1
Collecting GanodermaScreening 100kg of spore, performing physical wall breaking treatment, adding 1/5 weight parts of water, adding a small amount of water for multiple times, making into strip-shaped granule by spiral granulator, naturally drying at room temperature, adding CO2Supercritical extraction apparatus by CO2Supercritical extraction at 25MPa and 30 deg.c for 2 hr to extract liposoluble components.
Adding 10 times of water into the degreased wall-broken ganoderma lucidum spore particles, heating at 100 ℃, preserving heat for 2 hours, filtering, adding 8 times of water into filter residues, heating at 100 ℃, preserving heat for 2 hours, filtering, combining filtrates, concentrating to a specific gravity of 1.15-1.23, adding 3 times of edible ethanol with a volume of 95%, stirring, standing overnight, taking out precipitates, adding a small amount of water, cleaning, and drying at low temperature to obtain 5kg of extract; the precipitate has polysaccharide content of 44.5%, polypeptide content of 25.4%, and is much higher than that of common Ganoderma spore extract.
Example 2
Selecting Ganoderma spore 100kg, performing physical wall breaking treatment, adding 1/5 weight parts of water, adding a little for several times, making into strip-shaped granule by spiral granulator, naturally drying at room temperature, adding CO2Supercritical extraction apparatus by CO2Supercritical extraction at 25MPa and 30 deg.c for 2 hr to extract liposoluble components.
Adding 8 times of water into the degreased wall-broken ganoderma lucidum spore particles, heating at 100 ℃, preserving heat for 2 hours, filtering, adding 6 times of water into filter residues, heating at 100 ℃, preserving heat for 2 hours, filtering, combining filtrates, concentrating to a specific gravity of 1.15-1.23, adding 2 times of edible ethanol with a volume of 95%, stirring, standing overnight, taking out precipitates, adding a small amount of water, cleaning, and drying at low temperature to obtain 5kg of extract; the precipitate contains polysaccharide 40.5% and polypeptide 21.2%, and is higher than common Ganoderma spore extract.
Test examples
The polysaccharide peptides of Ganoderma spore powder used in the following experimental examples were prepared by the method of example 1.
1. The tested drugs are: EXAMPLE 1 preparation of polysaccharide peptide from Ganoderma spore powder
The preparation method comprises the following steps: the dosage of polysaccharide peptide of Ganoderma spore powder is 0.85g/kg*Every day, 442mg of the ganoderma lucidum spore powder polysaccharide peptide prepared in example 1 is precisely weighed, 5.2mL of CMC-Na is added, the mixture is fully dissolved by vortex, and the administration is carried out by intragastric administration every day, wherein the administration volume is 0.2 mL.
2. Test cell
Mouse Lewis lung carcinoma cells were derived from ATCC; culturing the cells in a DMEM medium containing 10% fetal bovine serum; cultured in a constant temperature incubator containing 5% CO2 at 37 ℃.
3. Laboratory animal
Strain and grade: SPF grade C57BL/6 mouse manufacturer: changzhou Kavensisi; the week age is as follows: sex 5-6 weeks: and (4) a female.
4. Experiment grouping
5. Experimental methods
Mice were collected with Lewis lung carcinoma cells and adjusted to a density of 4X 106/mL, and the cells were inoculated subcutaneously into axilla of mice at a rate of 0.2 mL/mouse, and on day 5 after inoculation, the mice were randomly divided into 12 groups and started.
Dosing was performed according to the above groups with 15 days of continuous intervention. The experimental period is 20 days; observing the mice during the experiment, starting the open field experiment, the tail suspension experiment, the forced swimming experiment and other ethological experiments when the activities are reduced, the skin color is dark, and the weight is reduced, and recording the related indexes; the animals were sacrificed at the end of the experiment, the tumors were removed, weighed and photographed; taking liver, instantly freezing with liquid nitrogen, storing at-80 deg.C, and measuring hepatic glycogen content with kit.
6. Results of the experiment
6.1 tumor weight and tumor inhibition Rate
When the experiment is finished, weighing the animals, stripping off tumors, weighing the tumors, and calculating the tumor inhibition rate; as shown in Table 1The weight average of tumors of each administration group is reduced very significantly compared with that of Lewis lung cancer group (***p<0.001); the tumor inhibition rate results show that all the medicines can effectively inhibit the Lewis lung cancer cell proliferation in vivo.
Table 1 tumor inhibition rate (N10, data Mean ± SD) for each administration group
Note: compared with the Lewis lung cancer group,***p<0.001。
6.2 mental fatigue test results
The mouse open field experiment can be used as one of evaluation criteria of mental fatigue; observing the activity of the mouse in the middle lattice of the open box, wherein the activity comprises the stay times in the middle lattice, the crossing times among the squares, namely the times of spanning more than three claws into the adjacent lattice, and the standing time, namely the time of more than 1cm above the ground for two forelimbs; the more the number of times of center stay, the more mental fatigue; the results in Table 3 show that the number of open field center stays of the mice in the Lewis lung cancer group is significantly higher than that in the blank control group: (*p<0.05); after the prognosis of drug administration, the number of stay times in the open field center of each group of mice is obviously lower than that of the Lewis lung cancer group: (###p<0.001), mental fatigue is remarkably relieved.
Table 2 number of stay times in open field (N10, data Mean ± SD) for each group of mice
| Group of | Number of stay in center of open field |
| Blank control group | 30.20±4.96 |
| Lewis lung cancer group | 45.50±21.96* |
| Ganoderma lucidum spore powder polysaccharide peptide group + Lewis lung cancer | 22.50±8.86### |
| Ganoderma lucidum spore powder polysaccharide peptide, cyclophosphamide group and Lewis lung cancer | 18.63±9.61### |
| Cyclophosphamide and Lewis lung carcinoma | 19.50±11.23### |
Note: compared with the blank control group, the composition of the composition,*p<0.05; compared with the Lewis lung cancer group,###p<0.001。
6.3 physical fatigue test results
The mouse tail suspension experiment can be used as one of evaluation standards of physical fatigue; fixing a 2cm part of the tail end of each mouse to enable the mouse to be in an inverted hanging state, enabling the head of each mouse to be 5-6 cm away from the ground, separating the sight lines of adjacent animals by using a wood board, adapting for 1min, and recording the accumulated motionless time of each mouse within 5 min; the shorter the struggling time of the mouse is, the more fatigue the mouse is; the results in Table 3 show that the ratio of struggling time of Lewis lung cancer mice is significantly lower than that of the blank control group (**p<0.01); after drug intervention, the struggling time ratio of each group of mice is obviously higher than that of Lewis lung cancer group: (#p<0.05), the fatigue of physical strength is relieved to a certain extent.
In addition, the exhaustive swimming test can also be used as another evaluation standard of physical fatigue; preparing a weight equivalent to 7% of the weight of a mouse, adhering the weight to the tail of the mouse by using a medical adhesive tape, putting the mouse into a swimming box with the water temperature of 25 +/-1 ℃ for swimming test until the force is exhausted and the mouse cannot sink, and recording the elapsed time; the shorter the mouse moves in water, the more fatigued the mouse is; the results are shown in Table 4, the struggling time of Lewis lung cancer miceThe ratio is significantly lower than that of a blank control group (***p<0.001); after the drug is administered for a dry dose, except for the cyclophosphamide group (*p>0.05), the struggling time ratio of each group of mice is obviously higher than that of Lewis lung cancer group: (#p<0.05), physical fatigue is remarkably relieved; because cyclophosphamide has multi-system toxicity, the physical fatigue of experimental mice of the cyclophosphamide group is presumed to be caused by the accumulated toxicity of the cyclophosphamide; and the other combination groups can effectively improve the physical fatigue of the mice.
Table 3 ratio of struggle time for suspending tail of each group of mice (N10, data Mean ± SD)
| Group of | Ratio of struggle time in hanging tail (%) |
| Blank control group | 91.44±8.01 |
| Lewis lung cancer group | 75.72±12.14** |
| Ganoderma lucidum spore powder polysaccharide peptide group + Lewis lung cancer | 91.85±8.16# |
| Ganoderma lucidum spore powder polysaccharide peptide (high) + cyclophosphamide group + Lewis lung cancer | 95.41±4.14# |
| Cyclophosphamide and Lewis lung carcinoma | 89.68±7.74# |
Note: compared with the blank control group, the composition of the composition,**p<0.01; compared with the Lewis lung cancer group,#p<0.05。
table 4 time to stop struggle for each group of mice to swim exhaustively (N10, data Mean ± SD)
| Group of | Time(s) for stopping struggle of mouse exhaustion swimming |
| Blank control group | 238.00±45.12 |
| Lewis lung cancer group | 113.30±16.40*** |
| Ganoderma lucidum spore powder polysaccharide peptide group + Lewis lung cancer | 211.50±43.33# |
| Ganoderma lucidum spore powder polysaccharide peptide (high) + cyclophosphamide group + Lewis lung cancer | 188.30±70.22# |
| Cyclophosphamide and Lewis lung carcinoma | 99.30±17.52 |
Note: in comparison to the blank set, the data is,***p<0.001; compared with the Lewis lung cancer group,#p<0.05。
6.4 hepatic glycogen content
Fatigue and drowsiness are insulin resistanceThe important performance of (2); the essence of hepatic insulin resistance is that insulin has a reduced ability to inhibit endogenous glucose production and reduced hepatic glycogen synthesis; therefore, the degree of the drug for relieving fatigue and drowsiness can be effectively reflected by measuring the content of hepatic glycogen; liver glycogen content determination experiment results shown in Table 5 show that the liver glycogen content of mice in Lewis lung cancer group is very significantly lower than that of mice in blank control group***p<0.001); after the drug is taken for prognosis, the liver glycogen content of a plurality of groups of mice is remarkably increased (##p<0.01), the cyclophosphamide group has certain rising effect when used alone.
Table 5 liver glycogen content (N10, data Mean ± SD) of each group of mice
Note: in comparison to the blank set, the data is,***p<0.001; compared with the Lewis lung cancer group,##p<0.01，###p<0.001。
each administration group can obviously inhibit the proliferation of Lewis lung cancer subcutaneous transplantation tumor; the administration combination can obviously relieve mental fatigue of the mice with cancer-related fatigue and also has a certain relieving effect on physical fatigue of the mice with cancer-related fatigue; the Ganoderma spore glycopeptide has effect in improving tumor-bearing mice with cancer-induced fatigue.
The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above-mentioned embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the invention may be made by those skilled in the art without departing from the principle of the invention.
Claims (10)
1. Application of Ganoderma spore powder polysaccharide peptide in preparing medicine for treating cancer-induced fatigue is provided.
2. The use of claim 1, wherein the polysaccharide peptide of Ganoderma lucidum spore powder has a polysaccharide content of 40% or more and a polypeptide content of 20% or more.
3. The use according to claim 1, wherein the method for preparing the polysaccharide peptide from ganoderma lucidum spore powder comprises the following steps:
(1) screening plump ganoderma lucidum spores for wall breaking;
(2) adding water into wall-broken Ganoderma spore powder, making into strip granule, drying at low temperature until water content is less than 6%, and adding CO2Supercritical extraction of fat-soluble components;
(3) adding water into defatted wall-broken Ganoderma spore granule, maintaining the temperature at 90-100 deg.C, filtering, repeatedly treating the residue for 1-3 times, mixing filtrates, concentrating, adding 95% edible ethanol, stirring, standing overnight, collecting precipitate, adding anhydrous ethanol, cleaning, and oven drying at low temperature.
4. The use according to claim 3, wherein in the step (1), the polysaccharide and the protein of the sporoderm-broken ganoderma lucidum spore powder are respectively not less than 1.0% and not less than 1.0%.
5. The use according to claim 3, wherein in the step (2), the amount of water is 1/5-1/4% by weight of the sporoderm-broken Ganoderma spore powder.
6. Use according to claim 3, wherein in step (2), CO is2The supercritical extraction conditions comprise extraction pressure of 25-28 MPa, extraction temperature of 25-30 ℃ and extraction time of 1-2 hours.
7. The use according to claim 3, wherein in step (2), the extraction pressure is 25MPa, the extraction temperature is 30 ℃, and the extraction time is 2 hours.
8. The application of the method as claimed in claim 3, wherein in the step (3), 8-10 times of water by weight is added into the defatted wall-broken ganoderma lucidum spore particles, the mixture is heated at 90-100 ℃, the mixture is filtered after heat preservation for 1-3 hours, 6-8 times of water by weight is added into filter residues, the mixture is heated at 90-100 ℃, the heat preservation is carried out for 1-3 hours, and the filter liquor is filtered and combined.
9. The use as claimed in claim 3, wherein in step (3), the filtrates are combined, concentrated to a specific gravity of 1.15-1.23, added with 95% edible ethanol 2-3 times the volume, stirred and left overnight to obtain a precipitate.
10. The use according to claim 3, wherein in the step (3), the drying temperature is 30-40 ℃.
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