Compound ganoderma lucidum spore oil for improving cancer-induced fatigue and preparation method and application thereof
阅读说明：本技术 改善癌因性疲乏的复方灵芝孢子油及其制备方法和应用 (Compound ganoderma lucidum spore oil for improving cancer-induced fatigue and preparation method and application thereof ) 是由 冯鹏 胡浪 彭雅娟 陈晓红 华克伟 于 2021-10-14 设计创作，主要内容包括：本发明公开了一种改善癌因性疲乏的复方灵芝孢子油及其制备方法和应用,该复方灵芝孢子油是通过以下方法制备得到的：先将灵芝细粉和破壁灵芝孢子粉与分别制粒、干燥,得到灵芝子实体颗粒和破壁灵芝孢子粉颗粒,将灵芝子实体颗粒和破壁灵芝孢子粉颗粒按照重量比1-2：1-2范围混合后进行CO2超临界萃取,采用的萃取条件：萃取釜温度38～42℃,萃取釜压力30～35MPa,萃取时间3～4h,收集萃取液。发明方法得到的产品具有显著的改善癌因性疲乏的药理活性,且将灵芝子实体和破壁灵芝孢子颗粒按照一定的比例混合萃取可以起到协同增效的作用。(The invention discloses a compound ganoderma lucidum spore oil for improving cancer-induced fatigue and a preparation method and application thereof, wherein the compound ganoderma lucidum spore oil is prepared by the following steps: respectively granulating and drying the fine ganoderma powder and the wall-broken ganoderma spore powder to obtain ganoderma fruit body granules and wall-broken ganoderma spore powder granules, and mixing the ganoderma fruit body granules and the wall-broken ganoderma spore powder granules according to a weight ratio of 1-2: 1-2, and performing CO2 supercritical extraction under the following extraction conditions: the temperature of the extraction kettle is 38-42 ℃, the pressure of the extraction kettle is 30-35 MPa, the extraction time is 3-4 h, and the extract liquor is collected. The product obtained by the method has the pharmacological activity of obviously improving cancer-induced fatigue, and the ganoderma lucidum fruiting body and the wall-broken ganoderma lucidum spore particles are mixed and extracted according to a certain proportion to play a role in synergy.)
1. The compound ganoderma lucidum spore oil for improving cancer-induced fatigue is characterized by being prepared by the following steps: respectively granulating and drying the fine ganoderma powder and the wall-broken ganoderma spore powder to obtain ganoderma fruit body granules and wall-broken ganoderma spore powder granules, and mixing the ganoderma fruit body granules and the wall-broken ganoderma spore powder granules according to a weight ratio of 1-2: 1-2 range mixing followed by CO2Supercritical extraction, wherein the adopted extraction conditions are as follows: the temperature of the extraction kettle is 38-42 ℃, the pressure of the extraction kettle is 30-35 MPa, the extraction time is 3-4 h, and the extract liquor is collected.
2. The compound ganoderma lucidum spore oil for improving cancer-induced fatigue as claimed in claim 1, wherein the ratio of ganoderma lucidum fruiting body particles to wall-broken ganoderma lucidum spore powder particles is 1: 1 and mixing.
3. The compound ganoderma lucidum spore oil for improving cancer-induced fatigue as claimed in claim 1, wherein the compound ganoderma lucidum spore oil is characterized by CO2Soaking the mixture of Ganoderma fruiting body granule and wall-broken Ganoderma spore powder granule in 95% ethanol for 1.5-2.5 hr before supercritical extraction.
4. The preparation method of the compound ganoderma lucidum spore oil for improving cancer-induced fatigue as claimed in claim 1, characterized in that the method comprises the following steps: respectively granulating and drying the fine ganoderma powder and the wall-broken ganoderma spore powder to obtain ganoderma fruit body granules and wall-broken ganoderma spore powder granules, and mixing the ganoderma fruit body granules and the wall-broken ganoderma spore powder granules according to a weight ratio of 1-2: 1-2 range mixing followed by CO2Supercritical extraction, wherein the adopted extraction conditions are as follows: the temperature of the extraction kettle is 38-42 ℃, the pressure of the extraction kettle is 30-35 MPa, the extraction time is 3-4 h, and the extract liquor is collected.
5. The method for preparing compound ganoderma lucidum spore oil for improving cancer-induced fatigue as claimed in claim 4, wherein the ratio of ganoderma lucidum fruiting body granules to wall-broken ganoderma lucidum spore powder granules is 1: 1 and mixing.
6. The method for preparing compound Ganoderma lucidum spore oil for improving cancer-induced fatigue as claimed in claim 4, wherein the CO is2Soaking the mixture of Ganoderma fruiting body granule and wall-broken Ganoderma spore powder granule in 95% ethanol for 1.5-2.5 hr before supercritical extraction.
7. The method for preparing compound ganoderma lucidum spore oil for improving cancer-induced fatigue as claimed in claim 6, wherein the CO is2Soaking the mixture of Ganoderma fruiting body granule and wall-broken Ganoderma spore powder granule with 95% ethanol for 2 hr before supercritical extraction.
8. The use of the compound ganoderma lucidum spore oil of claim 1 in the preparation of a medicament for improving cancer-related fatigue.
Technical Field
The invention belongs to the technical field of pharmacy, and particularly relates to compound ganoderma lucidum spore oil capable of improving cancer-induced fatigue, and a preparation method and application thereof.
Background
Cancer-induced Fatigue (CRF) refers to a Fatigue sensation caused by a tumor or tumor treatment, and a patient is stressed and painful for a long time, and may have symptoms such as weakness, difficulty in concentrating attention, poor activity endurance, lack of interest, and reduced motivation. Common to tumor patients receiving radiotherapy, chemotherapy and biotherapy, about 80% of patients receiving chemotherapy/radiotherapy may develop fatigue symptoms, seriously affecting the quality of life of the patient. The traditional Chinese medicine has unique advantages in the aspect of improving fatigue symptoms, has wide research on fatigue phenomena in various chronic diseases, but has not been frequently researched in cancer-induced fatigue.
Ganoderma lucidum is neutral in nature and sweet in taste, is a rare traditional Chinese medicine in China, and is long-term regarded as a precious Chinese herbal medicine for nourishing, strengthening the body, consolidating the constitution and strengthening the body resistance. Through a large number of clinical researches, the ganoderma lucidum has different degrees of curative effects on immunoregulation, auxiliary tumor suppression, neurasthenia, hyperlipidemia, coronary heart disease, angina pectoris, arrhythmia, liver protection, dyspepsia, tracheitis and the like.
The medicinal effect of the ganoderma lucidum spore powder is gradually recognized and accepted by consumers in nearly two decades. Along with the development of various technologies, the yield of ganoderma lucidum spore powder is higher and higher, the related research is increased gradually, the ganoderma lucidum spore oil is oily lipid extracted from the wall-broken ganoderma lucidum spores through a supercritical carbon dioxide fluid extraction technology, and a plurality of active ingredients of the ganoderma lucidum spores are integrated.
The traditional Chinese medicine considers that the disease condition is simple, a medicine with strong pertinence is selected to obtain the curative effect, and the traditional Chinese medicine accords with the requirements of simplicity, convenience, cheapness and experience and is convenient to use and popularize. However, if the condition of an illness is complicated, a single medicine cannot achieve the treatment requirements of primary and secondary differentiation and comprehensive consideration, more than two medicines need to be used simultaneously, certain interaction occurs between the medicines, and if some medicines generate synergistic effect to improve the curative effect, the traditional Chinese medicine components are complicated, and the possibility of mutually conflicting effective effects in mixed use needs to be fully noticed during clinical medication.
The ganoderma triterpene is one of main effective components in ganoderma sporocarp and ganoderma spore powder, has wide pharmacological action, and brings great limitation to deep research and clinical application of the triterpene due to low content and difficult separation of the ganoderma triterpene in ganoderma. The invention mixes the ganoderma lucidum sporocarp and the ganoderma lucidum spore powder and carries out supercritical CO treatment2Extracting to obtain compound Ganoderma spore oil, and exploring its effect in improving cancer-induced fatigue.
Disclosure of Invention
The invention aims to overcome the defects and provide the compound ganoderma lucidum spore oil for improving cancer-induced fatigue.
The invention also aims to provide a preparation method of the compound ganoderma lucidum spore oil for improving cancer-induced fatigue.
The purpose of the invention is realized by the following modes:
a compound ganoderma lucidum spore oil for improving cancer-induced fatigue is prepared by the following steps: respectively granulating and drying the fine ganoderma powder and the wall-broken ganoderma spore powder to obtain ganoderma fruit body granules and wall-broken ganoderma spore powder granules, and mixing the ganoderma fruit body granules and the wall-broken ganoderma spore powder granules according to a weight ratio of 1-2: 1-2 range mixing followed by CO2Supercritical extraction, wherein the adopted extraction conditions are as follows: the temperature of the extraction kettle is 38-42 ℃, and the pressure of the extraction kettle is 30-c35MPa, extracting for 3-4 h, and collecting the extract.
The ganoderma lucidum fruiting body particles and the wall-broken ganoderma lucidum spore powder particles are mixed according to the weight ratio of 1: 1 and mixing.
CO as described above2Soaking the mixture of Ganoderma fruiting body granule and wall-broken Ganoderma spore powder granule in 95% ethanol for 1.5-2.5 hr before supercritical extraction.
The extracts obtained after soaking and not soaking are respectively subjected to ganoderma triterpene determination, and the detection method comprises the following steps:
precisely weighing 10mg of oleanolic acid reference substance dried at 105 ℃ to constant weight, placing in a 50ml volumetric flask, adding chloroform to dissolve and dilute to scale, and obtaining the oleanolic acid standard reference substance solution.
Control solution
Precisely sucking standard oleanolic acid reference substance solution 0.1, 0.2, 0.4, 0.6, 0.8ml, respectively placing into 10ml colorimetric tubes, heating to volatilize solvent, adding 80g/L vanillin solution 0.5ml and sulfuric acid solution 5.0ml, mixing, keeping temperature in 60 deg.C water bath for 30min, taking out, and standing in cold water bath for 15 min. In a 1cm cuvette, the reagent blank solution is used as a reference for zero adjustment, the absorbance is measured at the wavelength of 500nm, and the absorbance is used for regression of the concentration to draw a standard curve.
2. Sample solution
Weighing 120-150 mg of un-soaked extract and soaked extract samples, and dissolving in a 100ml volumetric flask by adding chloroform.
Precisely sucking 0.2ml to 10ml of sample solution into a colorimetric tube, operating according to the item of the reference solution, namely heating to volatilize the solvent, measuring absorbance, and calculating the content.
3. Computing
In the formula:
w-total triterpene content in the sample%
C-oleanolic acid content, mg/ml, from Standard Curve
Volume of V-color solution, ml
F-dilution multiple
m-mass of sample, mg
4. As a result:
the content of ganoderma triterpene in the extract after soaking is higher, and the ganoderma triterpene extract is preferably soaked in 95 percent ethanol for 1.5 to 2.5 hours.
The preparation method of the compound ganoderma lucidum spore oil for improving cancer-induced fatigue comprises the following steps:
respectively granulating and drying the fine ganoderma powder and the wall-broken ganoderma spore powder to obtain ganoderma fruit body granules and wall-broken ganoderma spore powder granules, and mixing the ganoderma fruit body granules and the wall-broken ganoderma spore powder granules according to a weight ratio of 1-2: 1-2 range mixing followed by CO2Supercritical extraction, wherein the adopted extraction conditions are as follows: the temperature of the extraction kettle is 38-42 ℃, the pressure of the extraction kettle is 30-35 MPa, the extraction time is 3-4 h, and the extract liquor is collected. Preferably, the weight ratio of the ganoderma lucidum fruiting body particles to the wall-broken ganoderma lucidum spore powder particles is 1: 1 and mixing. Preferably in CO2Soaking the mixture of Ganoderma fruiting body granule and wall-broken Ganoderma spore powder granule in 95% ethanol for 1.5-2.5 hr before supercritical extraction.
The compound ganoderma lucidum spore oil has obvious effect in preparing the medicine for improving cancer-induced fatigue.
Compared with the prior art, the invention has the beneficial effects that: the product obtained by the method has the pharmacological activity of obviously improving cancer-induced fatigue, and the ganoderma lucidum fruiting body particles and the wall-broken ganoderma lucidum spore particles are mixed and extracted according to a certain proportion, so that the synergistic effect can be achieved.
Detailed Description
The invention is illustrated by the following specific examples:
example 1
1) Crushing the lucid ganoderma by using a crusher, and sieving the crushed lucid ganoderma by using a sieve with 40-50 meshes to obtain lucid ganoderma fine powder. Adding water 30 wt% of Ganoderma fine powder into Ganoderma fine powder, granulating in granulator, and vacuum drying at 60 deg.C for 2 hr to obtain Ganoderma granule.
2) Breaking cell wall of Ganoderma spore powder with cell wall breaking rate of 70%. Adding the wall-broken ganoderma lucidum spore powder into water with the weight of 30% of the wall-broken ganoderma lucidum spore powder, granulating, and drying in a vacuum drying oven at 60 ℃ for 2h to obtain the wall-broken ganoderma lucidum spore powder particles.
3) Mixing Ganoderma granule and wall-broken Ganoderma spore powder at equal ratio, placing into supercritical extraction equipment, soaking with 95% ethanol at the same weight for 2 hr, and subjecting the soaked mixture to CO2Supercritical extraction, wherein the extraction conditions are as follows: the temperature of an extraction kettle is 40 ℃, the pressure of the extraction kettle is 30MPa, the extraction time is 3 hours, extract liquor is collected to obtain compound ganoderma lucidum spore oil which is dark brown ethanol liquid containing oily substances, the ethanol is removed to obtain dark brown oily liquid, and the compound ganoderma lucidum spore oil is filtered, wherein the triterpene content in the compound ganoderma lucidum spore oil is 57.5 percent, and the extraction rate is 13.6 percent.
Example 2
1) Breaking cell wall of Ganoderma spore powder with cell wall breaking rate of 70%. Adding the wall-broken ganoderma lucidum spore powder into water with the weight of 30% of the wall-broken ganoderma lucidum spore powder, granulating, and drying in a vacuum drying oven at 60 ℃ for 2h to obtain the wall-broken ganoderma lucidum spore powder particles.
2) Placing the wall-broken Ganoderma spore powder into supercritical extraction equipment, soaking with 95% ethanol at the same ratio for 2 hr, and subjecting the soaked wall-broken Ganoderma spore powder to CO extraction2Supercritical extraction, wherein the extraction conditions are as follows: extracting at 40 deg.C under 30MPa for 3 hr, collecting extractive solution as brown ethanol liquid containing oily substance, removing ethanol to obtain brown oily liquid, and filtering. The triterpene content in the extract is 15.4 percent, and the extraction rate is 22.4 percent.
Example 3
1) Crushing the lucid ganoderma by using a crusher, and sieving the crushed lucid ganoderma by using a sieve with 40-50 meshes to obtain lucid ganoderma fine powder. Adding water 30 wt% of Ganoderma fine powder into Ganoderma fine powder, granulating in granulator, and vacuum drying at 60 deg.C for 2 hr to obtain Ganoderma granule.
2) Placing Ganoderma granule into supercritical extraction equipment, soaking with 95% ethanol at the same ratio for 2 hr, and adding the soaked Ganoderma granuleCO removal2Supercritical extraction, wherein the extraction conditions are as follows: extracting at 40 deg.C under 30MPa for 3 hr, collecting extractive solution as dark brown ethanol liquid containing pasty substance, removing ethanol to obtain dark brown pasty solid, and filtering. The triterpene content in the extract was 61.4%, and the extraction rate was 2.6%.
Example 4
1) Crushing the lucid ganoderma by using a crusher, and sieving the crushed lucid ganoderma by using a sieve with 40-50 meshes to obtain lucid ganoderma fine powder. Adding water 30 wt% of Ganoderma fine powder into Ganoderma fine powder, granulating in granulator, and vacuum drying at 60 deg.C for 2 hr to obtain Ganoderma granule.
2) Breaking cell wall of Ganoderma spore powder with cell wall breaking rate of 70%. Adding the wall-broken ganoderma lucidum spore powder into water with the weight of 30% of the wall-broken ganoderma lucidum spore powder, granulating, and drying in a vacuum drying oven at 60 ℃ for 2h to obtain the wall-broken ganoderma lucidum spore powder particles.
3) Mixing Ganoderma granule and wall-broken Ganoderma spore powder granule at a weight ratio of 1:2, placing into supercritical extraction equipment, soaking with 95% ethanol at the same weight ratio for 2 hr, and subjecting the soaked mixture to CO2Supercritical extraction, wherein the extraction conditions are as follows: the temperature of an extraction kettle is 40 ℃, the pressure of the extraction kettle is 30MPa, the extraction time is 3 hours, extract liquor is collected to obtain compound ganoderma lucidum spore oil which is dark brown ethanol liquid containing oily substances, the ethanol is removed to obtain dark brown oily liquid, and the compound ganoderma lucidum spore oil is filtered, wherein the triterpene content in the compound ganoderma lucidum spore oil is 44.2 percent, and the extraction rate is 17.5 percent.
Test example 1
The invention is further illustrated by the following pharmacodynamic experiments:
1. purpose of the experiment: the improvement effect of the compound ganoderma lucidum spore oil on tumor-bearing mice with cancer-induced fatigue is explored.
2. Experimental Material
2.1 test drug 1: compound ganoderma lucidum spore oil (prepared according to the method of example 1)
The preparation method comprises the following steps: the compound Ganoderma spore oil has high dose of 1g/kg day, medium dose of 0.67g/kg day, and low dose of 0.33g/kg day. The density of the compound ganoderma lucidum spore oil is 0.8g/mL, 0.325mL, 0.218mL and 0.107mL of the compound ganoderma lucidum spore oil are precisely measured respectively, 2.275mL, 2.382mL and 2.493mL of soybean oil are added respectively, the mixture is fully and uniformly mixed, the intragastric administration is carried out every day, and the administration volume is 0.2 mL.
Test drug 2: ganoderma spore oil (prepared according to example 2)
The preparation method comprises the following steps: the Ganoderma spore oil dosage is 0.67g/kg day. The density of the ganoderma lucidum spore oil is 0.8g/mL, 0.218mL of ganoderma lucidum spore oil is precisely measured, 2.382mL of soybean oil is added, the mixture is fully and uniformly mixed, the administration is performed by intragastric administration every day, and the administration volume is 0.2 mL.
Test drug 3: ganoderma lucidum extract (prepared according to example 3)
The preparation method comprises the following steps: the dose of Ganoderma extract is 0.67g/kg day. Precisely weighing 0.174g of Ganoderma extract, adding 2.382mL of soybean oil, mixing, and intragastrically administering with administration volume of 0.2mL every day.
Test drug 4: compound ganoderma lucidum spore oil (prepared according to the method of example 4)
The preparation method comprises the following steps: the dosage of compound Ganoderma spore oil is 0.67g/kg day. The density of the compound ganoderma lucidum spore oil is 0.8g/mL, 0.218mL of the compound ganoderma lucidum spore oil is precisely measured, 2.382mL of soybean oil is added, the mixture is fully and uniformly mixed, the intragastric administration is carried out every day, and the administration volume is 0.2 mL.
Molding medicine: cyclophosphamide for injection (amdane) manufacturer: baxter Oncology GmbH lot number: 5N053A
The preparation method comprises the following steps: cyclophosphamide was administered at a dose of 30mg/kg day. 1 bottle (0.2g) of cyclophosphamide for injection is fully dissolved in 10mL of sterile normal saline, each tube is subpackaged with 2.7mL and stored at the temperature of minus 20 ℃, fully frozen before use, 15.3mL of sterile normal saline is added to dilute to the administration concentration, and the intraperitoneal injection is carried out every other day with the administration volume of 0.2 mL.
2.2 Experimental reagents
Liver/muscle glycogen assay kit (colorimetric): the Nanjing is built into a product of bioengineering institute, and the product is A043-1-1; concentrated sulfuric acid: nanjing reagent factory export, product number: c0680150225
2.3 test cells
Mouse Lewis lung carcinoma cells were derived from ATCC. The cells were cultured in DMEM medium containing 10% fetal bovine serum.Culturing at 37 deg.C with 5% CO2The constant temperature incubator.
2.4 Experimental animals
Strain and grade: SPF grade C57BL/6 mouse manufacturer: license number of Changzhou Kavens laboratory animals GmbH: SCXK (threo) 2016-; the week age is as follows: sex 5-6 weeks: a female; feed: the mice maintain the feed, and the breeding conditions of the Qinglong mountain animal breeding farm in Jiangning district of Nanjing are as follows: air-conditioned room with temperature of 18-24 deg.C and relative humidity of 70%
2.5 Experimental apparatus
An enzyme-labeling instrument: molecular Devices, Inc water bath: open field experimental system of mouse of Changzhou country china electric appliances Limited: CleverSys, Inc mouse tail suspension experimental system: CleverSys, Inc mouse forced swimming experimental system: CleverSys, Inc
3. Experimental methods
3.1 Experimental groups
3.2 Experimental methods
Collecting mouse Lewis lung cancer cells, and adjusting the density to 4 × 106At each 0.2 mL/mouse axilla, the cell suspension was injected subcutaneously for inoculation, and on day 5 after inoculation, administration was started according to the above groups for 15 days with continuous intervention. The experimental period was 20 days. The mice are observed during the experiment period, and the open field experiment, the tail suspension experiment, the forced swimming experiment and other ethological experiments are started when the activity is reduced, the skin color is dark, and the weight is reduced, and the relevant indexes are recorded. Animals were sacrificed at the end of the experiment, 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.
3.3 detection of indicators
3.3.1 animal body weight: initial body weights were recorded for mice, and body weights were administered on days 1, 5, 10, and 15, respectively.
3.3.2 mental fatigue: the mouse open field experiment can be used as one of evaluation criteria of mental fatigue. The activity of the mouse in the middle lattice of the open box is observed, including 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.
3.3.3 fatigue: the mouse tail suspension experiment can be used as one of evaluation criteria of physical fatigue. Fixing the 2cm position 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 exhaustion swimming test can also be used as one of the evaluation criteria of physical fatigue. Preparing a weight equivalent to 7 percent of the weight of the 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.
3.3.4 tumor weight and tumor inhibition rate: and respectively recording the tumor weight of each group, comparing with a Lewis lung cancer group, and calculating the tumor inhibition rate. The tumor inhibition rate calculation formula is as follows: tumor inhibition (%) was 100- (mean tumor weight in dosing group/mean tumor weight in Lewis lung cancer group)
*100％
3.3.5 hepatic glycogen content: liver glycogen content was measured separately for each group of mice.
4. Results of the experiment
4.1 tumor weight and tumor inhibition rate
At the end of the experiment, animals were weighed, tumors were stripped, tumor weights were weighed, and tumor inhibition rates were calculated. As shown in Table 1, the weight average of tumors was very significantly decreased in each administration group compared to that of Lewis lung cancer group (***p<0.001). The tumor inhibition rate results show that all the medicines can effectively inhibit the proliferation of Lewis lung cancer cells in vivo, and the compound preparation in example 1 has the best effect.
Table 1. tumor inhibition rate (N10, data Mean ± SD) for each administration group
Note: compared with the Lewis lung cancer group,***p<0.001。
4.2 mental fatigue test results
The staying times of the mouse in the open field center can be used as one of the measures of mental fatigue. The more the number of central stays, the more mental strain. The results in Table 2 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. In example 1, the combination of three doses of compound ganoderma lucidum spore oil and cyclophosphamide is obvious.
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*
Example 1 Compound Ganoderma spore oil (high) + Cyclophosphamide group + Lewis lung carcinoma
10.50±4.12###
Example 1 Compound Ganoderma spore oil (Medium) + Cyclophosphamide group + Lewis lung carcinoma
13.40±9.11###
Example 1 Compound Ganoderma spore oil (Low) + Cyclophosphamide group + Lewis Lung cancer
12.45±10.37###
Example 2 Ganoderma lucidum spore oil + Cyclophosphamide group + Lewis lung carcinoma
18.63±9.61###
Example 3 Ganoderma lucidum extract + Cyclophosphamide group + Lewis Lung
16.70±7.36###
Example 4 Compound Ganoderma spore oil (Medium) + Cyclophosphamide group + Lewis lung carcinoma
21.56±9.10###
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。
4.3 physical fatigue test results
The mouse tail suspension experiment can be used as one of the evaluation criteria of physical fatigue. The shorter the struggling time of the mouse, 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, exhaustive swimming tests can also be used as another assessment criterion for physical fatigue. The shorter the mouse is moving in water, the more fatigued the mouse is. Results Table 4 shows when Lewis lung carcinoma mice struggledThe space 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), the fatigue of physical strength is obviously relieved. Because cyclophosphamide has multiple systemic toxicities, we speculate that physical fatigue of experimental mice in the cyclophosphamide group is caused by accumulated toxicity of cyclophosphamide. And the other combination groups can effectively improve the physical fatigue of the mice.
Table 3 ratio of struggle time for hanging tail (N10, data Mean ± SD) for each group of mice
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 struggle for each group of mice to swim exhaustively (N10, data Mean ± SD)
Note: in comparison to the blank set, the data is,***p<0.001; compared with the Lewis lung cancer group,#p<0.05。
4.4 liver glycogen assay
Fatigue and drowsiness are important manifestations of insulin resistance. The essence of hepatic insulin resistance is that insulin's ability to inhibit endogenous glucose production is diminished and hepatic glycogen synthesis is reduced. Therefore, the degree of the drug relieving fatigue and drowsiness can be effectively reflected by measuring the content of hepatic glycogen. The liver glycogen content determination experiment results (Table 5) show that the liver glycogen content of mice in the Lewis lung cancer group is extremely lower than that of mice in the blank control group (***p<0.001); drug administration for prognosis of liver glycogen in multiple groups of miceThe content is remarkably increased (##p<0.01), wherein the compound ganoderma lucidum spore oil (high) + cyclophosphamide group and the compound ganoderma lucidum spore oil (low) + cyclophosphamide group in the example 1 have the most obvious rising effect, and the cyclophosphamide group has a certain rising effect when being used singly.
Table 5 liver glycogen content (N10, data Mean ± SD) of each group of mice
Group of
Mouse liver glycogen content (mg/g tissue)
Blank control group
5.01±1.306
Lewis lung cancer group
2.185±0.7102***
Example 1 Compound Ganoderma spore oil (high) + Cyclophosphamide group + Lewis lung carcinoma
7.833±2.675###
Example 1 Compound Ganoderma spore oil (Medium) + Cyclophosphamide group + Lewis lung carcinoma
2.239±1.094
Example 1 Compound Ganoderma spore oil (Low) + Cyclophosphamide group + Lewis Lung cancer
7.463±1.867###
Example 2 Ganoderma lucidum spore oil + Cyclophosphamide group + Lewis lung carcinoma
3.8±0.9194##
Example 3 Ganoderma lucidum extract + Cyclophosphamide group + Lewis Lung cancer
3.546±2.309##
Example 4 Compound Ganoderma spore oil (Low) + Cyclophosphamide group + Lewis Lung cancer
3.955±0.6356##
Cyclophosphamide and Lewis lung carcinoma
5.696±1.987##
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。
4.5 conclusion of the experiment
The model of the experimental cancer-induced fatigue model is successfully modeled. Each administration group can obviously inhibit the proliferation of Lewis lung cancer subcutaneous transplantation tumor. The administration combination can obviously relieve mental fatigue of mice with cancer-related fatigue and has certain relieving effect on physical fatigue of the mice with cancer-related fatigue. The compound ganoderma lucidum spore oil of the embodiment 1 has the effect of improving the tumor-bearing mice with cancer-induced fatigue in each dose.
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