Method for establishing model for cancer-related fatigue caused by antimetabolite chemotherapy drug
By pre-administering a combination of lipid solvents and antimetabolite chemotherapeutic drugs to animals, a highly efficient antimetabolite chemotherapeutic drug-induced carcinogenic fatigue model was established, solving the problems of long experimental cycles and high animal mortality, and achieving simplified operation and efficient drug screening.
Patent Information
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- GUANGZHOU HANFANG PHARMA CO LTD
- Filing Date
- 2024-11-21
- Publication Date
- 2026-05-15
AI Technical Summary
Existing methods for establishing carcinogenic fatigue models induced by antimetabolite chemotherapy drugs suffer from problems such as excessively long experimental cycles, cumbersome procedures, and high animal mortality rates, and lack of unified detection indicators and standards.
Animals were given an oil-based solvent for 15–20 days before being injected intraperitoneally with antimetabolite chemotherapy drugs to establish a model. The third day after modeling was selected as the sampling point. The degree of fatigue was detected by rotarod fatigue test and exhaustive swimming test. Ganoderma lucidum spore oil was used as the solvent for the positive drug group.
It shortens the experimental cycle, reduces animal mortality, improves the success rate and operability of the model, effectively reflects cancer-related fatigue, and facilitates drug screening and evaluation.
Smart Images
Figure CN2024133395_15052026_PF_FP_ABST
Abstract
Description
A method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs Technical Field
[0001] This invention relates to the field of pharmaceutical technology, specifically to a method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs. Background Technology
[0002] Cancer-related fatigue (CRF) is one of the most common symptoms in cancer patients and a frequent side effect of cancer treatment. The National Comprehensive Cancer Network (NCCN) defines it as "a distressing, persistent, subjective feeling of physical, emotional, or cognitive fatigue or exhaustion that is disproportionate to recent activity, related to cancer or cancer treatment, and interferes with daily life." The International Classification of Diseases (ICD) describes symptoms of cancer-related fatigue as nonspecific weakness, frailty, general deterioration, somnolence, and fatigue. Statistics show that cancer-related fatigue symptoms occur in more than 75% of cancer patients.
[0003] Currently, chemotherapy remains one of the main methods of cancer treatment. Commonly used chemotherapy drugs include antibiotics, antimetabolites, alkaloids, and alkylating agents. However, while effectively killing tumor cells, chemotherapy also brings a series of side effects, including the development or exacerbation of cancer-related fatigue. Antimetabolites are a class of small-molecule compounds with molecular structures and functions similar to the metabolites in human nucleic acid synthesis. They exert their anti-cancer effects by affecting nucleic acid metabolism. When cells mistakenly ingest these chemotherapy drugs during normal metabolism, they can inhibit key enzymes in nucleic acid synthesis or bind to nucleic acids and embed themselves in nucleic acid molecules, producing abnormal coding, leading to inhibited DNA synthesis and ultimately cell death.
[0004] However, the mechanism of cancer-related fatigue has not yet been definitively established, and there are no drugs on the market specifically for treating cancer-related fatigue. There is also no unified standard for the establishment of cancer-related fatigue models induced by antimetabolites and for the detection indicators. The applicant used the methods reported in Qi Xiaoye et al. in "Regulatory Effect of Guipi Decoction on TNF-α Secretion in Chemotherapy-Related Fatigue Model" (Asia Pacific Traditional Medicine, 2019(1):13-15) and Sara E. Mahoney et al. in "Dietary Quercetin Reduces Chemotherapy-Induced Fatigue in Mice" (Integrative Cancer Therapies, 2014, Vol.13(5):417-424) to conduct experiments. It was found that there are still many shortcomings, such as: the experimental period after injection modeling is as long as 14 days, in which the former resulted in the death of experimental animals, while the latter required a higher dosage and more detections. Therefore, overcoming the current problems of excessively long experimental cycles, overly cumbersome testing procedures, and a large number of animal deaths, and establishing a representative and operable animal model of cancer-related fatigue is of great significance for the study of the mechanism of cancer-related fatigue and the screening of therapeutic drugs. Summary of the Invention
[0005] The purpose of this invention is to improve models that suffer from high rates of animal mortality during the experimental process and excessively long experimental cycles after modeling.
[0006] To address the aforementioned technical problems, this invention provides a method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs. This model is created by pre-administering an oil-based solvent to animals for an extended period, followed by intraperitoneal injection of antimetabolite chemotherapy drugs. The third day after model establishment is selected as the sampling time. Before sampling, the first drop time of the rotator and the number of drops within 10 minutes, as well as the time spent swimming to exhaustion, are selected as detection indicators.
[0007] The present invention adopts the following technical solution:
[0008] A method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs, the method comprising: administering an oil-based solvent to experimental animals for 15-20 days prior to modeling.
[0009] As a preferred embodiment of the method of the present invention, the oil-based solvent includes one or more of soybean oil, corn oil, olive oil, and sesame oil; the modeling includes administering antimetabolite chemotherapy drugs to the experimental animals.
[0010] As a preferred embodiment of the method of the present invention, the establishment method includes the following steps:
[0011] A. The experimental animals were randomly divided into groups. The experimental animals in the blank control group were injected intraperitoneally with sterile water, while the experimental animals in the model control group, solvent control group, and positive drug group were injected intraperitoneally with antimetabolite chemotherapy drugs once a day for 5 consecutive days. The experimental animals in the blank control group and the model control group were given sterile water in advance before the intraperitoneal injection. The experimental animals in the solvent control group were given the oil-based solvent in advance before the intraperitoneal injection, and the experimental animals in the positive drug group were given a mixture of the oil-based solvent and the positive drug for cancer-induced fatigue in advance before the intraperitoneal injection. The advance administration time was 15-20 days, once a day.
[0012] B. After 5 days of intraperitoneal injection, the animals in each group were assessed for fatigue using behavioral experiments.
[0013] Furthermore, the pre-administration method is gavage; the pre-administration in step A begins 15 to 20 days before the intraperitoneal injection of the experimental animals, once a day, and continues until the animal fatigue level is detected in step B.
[0014] Furthermore, the experimental animals include mice; the daily dose of intraperitoneal injection for each group of experimental animals is 40 mg / kg b.wt.; the antimetabolite chemotherapy drug is fluorouracil; the volume of pre-administered drugs for each group of experimental animals is 5 mL / kg b.wt.
[0015] Further, step B includes: on days 1 to 3 after the completion of 5 days of intraperitoneal injection in each group of experimental animals, behavioral experiments are used to detect the degree of animal fatigue; the behavioral experiments include one or more of the rotarod fatigue test and the exhaustive swimming test.
[0016] Furthermore, the method for the rotator fatigue test includes: placing the experimental animal on the rotator, starting the timer from when the animal is placed on the rotator, and recording the first fall time and the number of falls within 10 minutes.
[0017] Furthermore, the method of the exhaustion swimming experiment includes: attaching a lead weight of 5% of the body weight to the tail of the experimental animal, placing the experimental animal in a swimming pool, starting the timer from the moment of entry into the water, and recording the exhaustion swimming time as the standard when the experimental animal's head sinks into the water and cannot float to the surface within 5 seconds.
[0018] The drugs mentioned for cancer-related fatigue include Ganoderma lucidum spore oil.
[0019] The composition of the Ganoderma lucidum spore oil includes: triglyceride content >90%, ergosterol content 0%–0.5%, and no detected Ganoderma lucidum triterpenoids. Before administration, prepare a solution of the appropriate concentration using an oil-based solvent, and use immediately after preparation.
[0020] The positive drug group mentioned above can be divided into low, medium and high dose groups of Ganoderma lucidum spore oil. The doses of Ganoderma lucidum spore oil administered to the animals by gavage in each group were 0.6, 1.2 and 2.4 g / kg b.wt. per day, respectively.
[0021] The preferred experimental animals used are C57BL / 6J mice.
[0022] The present invention also provides a model established according to the above-mentioned method for establishing a carcinogenic fatigue model of antimetabolite chemotherapy drugs.
[0023] The present invention also provides an application of the above model in studying cancer-related fatigue caused by antimetabolite chemotherapy drugs and / or evaluating the efficacy of drug treatment for cancer-related fatigue.
[0024] The method of the present invention has the following advantages compared with the prior art:
[0025] 1. The carcinogenic fatigue model of antimetabolite chemotherapy drugs of the present invention overcomes the shortcomings of existing methods, which easily lead to a large number of animal deaths and make it difficult to obtain sufficient effective data, by pre-administering oil-based solvents for a longer period of time. Therefore, the present invention has the advantage of reducing costs and increasing efficiency.
[0026] 2. The present invention has a high modeling success rate and ensures that the animal is in the most suitable state for drug evaluation without affecting the overall fatigue of the animal, and can better reflect the situation of cancer-related fatigue in animals.
[0027] 3. The present invention significantly shortens the observation period of animal behavior after modeling, thereby simplifying the experiment and making it easier to operate.
[0028] 4. The model establishment method of the present invention can effectively reflect the fatigue caused by the clinical application of antimetabolite chemotherapy drugs, and can be used for the study of the pathogenesis of cancer-related fatigue and the screening and evaluation of anti-cancer fatigue drugs. Attached Figure Description
[0029] Figure 1 is a comparison chart of the exhaustion swimming time of the embodiment and the comparative model;
[0030] Figure 2 is a comparison chart of the number of animal deaths in the example and the comparative model. Detailed Implementation
[0031] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments and comparative examples.
[0032] Example 1
[0033] Establishment of a mouse model of cancer-related fatigue induced by fluorouracil and evaluation of therapeutic drugs
[0034] 1. Laboratory animals
[0035] C57BL / 6J mice, SPF grade, half male and half female, weighing 16-20g, a total of 96 mice, were housed in an SPF-grade animal room with free access to food, 12h light / 12h darkness, and an animal room temperature of 16-26℃ and relative humidity of 40%-70%.
[0036] 2. Experimental Materials
[0037] Modeling agent: Fluorouracil for injection.
[0038] Drug administration solvent: soybean oil.
[0039] Ganoderma lucidum spore oil: triglyceride content >90%, ergosterol content 0%–0.5%, and no Ganoderma lucidum triterpenoids detected. Prepare a solution of the appropriate concentration with soybean oil before administration; use immediately after preparation.
[0040] 3. Experimental Methods
[0041] (1) Establishment and administration of a fluorouracil-induced mouse model of cancer-related fatigue
[0042] Mice were randomly divided into 6 groups: blank control group, model control group, solvent control group, high-dose Ganoderma lucidum spore oil group, medium-dose Ganoderma lucidum spore oil group, and low-dose Ganoderma lucidum spore oil group, with 16 mice in each group, half male and half female. Animals in the blank control group and model control group were administered sterile water by gavage; animals in the solvent control group were administered solvent by gavage; animals in the low-, medium-, and high-dose Ganoderma lucidum spore oil groups were administered Ganoderma lucidum spore oil by gavage, with daily doses of 0.6, 1.2, and 2.4 g / kg, respectively. All groups were administered the drug once daily, with an administration volume of 5 mL / kg. The third day after model establishment was selected as the sampling point, and administration was continuous from 15 days before model establishment until the day before sampling, once daily.
[0043] Fifteen days after pre-administration, animals in the blank control group were injected intraperitoneally with sterile water, while animals in the other groups were injected intraperitoneally with fluorouracil at a dose of 40 mg / kg once daily for five consecutive days. On the third day after modeling, the animals were dissected and their tissues were collected.
[0044] (2) Rotating bar fatigue test
[0045] Experimental equipment and conditions: YLS-4D rotor fatigue tester (rotor diameter 30mm, length 60mm), 1-4 channels, rotation speed set to 30r / min, acceleration time set to 30 seconds.
[0046] Experimental procedure: On the first day after modeling, the mice were placed on a rotarod apparatus. Timing was started from the moment the mice were placed on the rotarod apparatus, and the time of the first fall and the number of falls within 10 minutes were recorded.
[0047] (3) Exhaustion Swimming Experiment
[0048] Experimental equipment and conditions: A constant temperature swimming pool was filled with water, with the water surface 5cm from the edge of the pool. The water temperature was set at 28℃ and the vortex was at its maximum. An acrylic sheet was used to divide the swimming pool into 4 areas. Four mice were placed in the 4 areas of the same swimming pool at the same time to conduct the experiment without interfering with each other.
[0049] Experimental procedure: On the second day after modeling, a lead weight of 5% of the mouse's body weight was attached to the mouse's tail, and the mouse was placed in the aforementioned swimming pool. Timing was started from the moment the mouse entered the water, and the time it took to swim to exhaustion was recorded as the standard for exhaustion when the mouse's head was submerged in the water and it could not float to the surface within 5 seconds.
[0050] 4. Experimental Results
[0051] During the experiment, except for the model control group where 4 animals died (mortality rate 25%), the animals in the other groups were generally in good condition and no deaths occurred. The specific times of death for the animals in the model control group were: 1 animal before the rotarod fatigue experiment, 2 animals before the exhaustive swimming experiment, and 1 animal after the exhaustive swimming experiment.
[0052] The animal weight measurement results are shown in Table 1. The measurement times were: before drug administration (on the day of pre-administration), before modeling (on the day of injection for modeling), and before tissue collection (on the day of dissection and tissue collection, i.e., the 3rd day after modeling). As can be seen from the results in Table 1, there was no significant difference in animal weight among the groups before modeling; compared with the blank control group, the animal weight in the model control group was significantly lower before tissue collection (P < 0.01), while there were no significant differences between the other groups and the model control group.
[0053] Table 1. Animal weight test results (g) Note: Compared with the blank control group, **P<0.01.
[0054] The results of the rotator fatigue test are shown in Table 2. As can be seen from the test results of each group in Table 2: compared with the blank control group, the first drop time of animals in both the model control group and the solvent control group was significantly shortened (P < 0.05), and the number of drops was significantly increased (P < 0.01); compared with the model control group, there was no significant difference in the first drop time and the number of drops in the solvent control group; compared with the solvent control group, the first drop time of animals in all dosage groups of Ganoderma lucidum spore oil was significantly prolonged (P < 0.05), and the number of drops in the medium and low dosage groups was significantly reduced (P < 0.01 or P < 0.05).
[0055] Table 2 Results of Rotor Fatigue Test Note: Compared with the blank control group, **P<0.01, *P<0.05; compared with the solvent control group, ## P < 0.01, # P < 0.05.
[0056] The results of the exhaustive swimming experiment are shown in Table 3. As can be seen from the test results in Table 3, compared with the blank control group, the exhaustive swimming time of animals in both the model control group and the solvent control group was significantly shortened (P < 0.01); compared with the model control group, there was no significant difference in the exhaustive swimming time of animals in the solvent control group; compared with the solvent control group, the exhaustive swimming time of animals in the medium and high dose groups of Ganoderma lucidum spore oil was significantly prolonged (P < 0.01).
[0057] Table 3 Results of the Exhaustion Swimming Test Note: Compared with the blank control group, **P < 0.01; compared with the solvent control group, ## P < 0.01.
[0058] Based on the detection data and differences between groups in Tables 1 to 3, the performance of the model in Example 1 within 3 days after modeling is analyzed as follows:
[0059] 1) Compared to the blank control group, the body weight of animals in each experimental group decreased to varying degrees after modeling, which can reflect the fatigue status of the animals to some extent. 2) The results of the rotarod fatigue test and the exhaustive swimming test showed that mice in both the model control group and the solvent control group exhibited significant fatigue in various behavioral indicators at the test time points. Ganoderma lucidum spore oil significantly alleviated the fatigue state of mice in the cancer-induced fatigue model, and various behavioral indicators could be restored to levels close to those of the blank control group, indicating that the cancer-induced fatigue model was successfully established. 3) Regarding the analysis of animal mortality, only the model control group experienced animal deaths, while the mice in the other groups, including the solvent control group, were in good condition and did not die. The underlying principle is speculated to be that compared to pre-administering sterile water (model control group), pre-administering a long-term oil-based solvent (solvent control group) increased the animals' tolerance to chemotherapy drugs, thereby reducing the mortality rate during the experiment and ensuring that the animals were in the most suitable state for drug evaluation without affecting the overall fatigue of the animals.
[0060] Example 2
[0061] Establishment of a mouse model of cancer-related fatigue induced by fluorouracil and evaluation of therapeutic drugs
[0062] 1. Laboratory animals
[0063] C57BL / 6J mice, SPF grade, half male and half female, weighing 16-20g, 64 mice in total. Housing environment: housed in an SPF grade animal room, with free access to food, 12h light / 12h darkness, animal room temperature 16-26℃, relative humidity 40%-70%.
[0064] 2. Experimental Materials
[0065] Modeling agent: Fluorouracil for injection.
[0066] Drug administration solvent: corn oil.
[0067] Ganoderma lucidum spore oil: triglyceride content >90%, ergosterol content 0%–0.5%, and no Ganoderma lucidum triterpenoids detected. Prepare a solution of the appropriate concentration with corn oil before administration; use immediately after preparation.
[0068] 3. Experimental Methods
[0069] (1) Establishment and administration of a fluorouracil-induced mouse model of cancer-related fatigue
[0070] Mice were randomly divided into four groups: blank control group, model control group, solvent control group, and medium-dose Ganoderma lucidum spore oil group, with 16 mice in each group (half male and half female). Animals in the blank control group and model control group were administered sterile water by gavage; animals in the solvent control group were administered solvent by gavage; and animals in the medium-dose Ganoderma lucidum spore oil group were administered 1.2 g / kg of Ganoderma lucidum spore oil by gavage daily. All groups were administered the drug once daily at a volume of 5 mL / kg. The third day after model establishment was selected as the sampling point, and administration was continuous from 15 days before model establishment until one day before sampling, once daily.
[0071] Fifteen days after pre-administration, animals in the blank control group were injected intraperitoneally with sterile water, while animals in the other groups were injected intraperitoneally with fluorouracil at a dose of 40 mg / kg once daily for five consecutive days. On the third day after modeling, the animals were dissected and their tissues were collected.
[0072] (2) Rotating bar fatigue test
[0073] Experimental equipment and conditions: YLS-4D rotor fatigue tester (rotor diameter 30mm, length 60mm), 1-4 channels, rotation speed set to 30r / min, acceleration time set to 30 seconds.
[0074] Experimental procedure: On the first day after modeling, the mice were placed on a rotarod apparatus. Timing was started from the moment the mice were placed on the rotarod apparatus, and the time of the first fall and the number of falls within 10 minutes were recorded.
[0075] (3) Exhaustion Swimming Experiment
[0076] Experimental equipment and conditions: A constant temperature swimming pool was filled with water, with the water surface 5cm from the edge of the pool. The water temperature was set at 28℃ and the vortex was at its maximum. An acrylic sheet was used to divide the swimming pool into 4 areas. Four mice were placed in the 4 areas of the same swimming pool at the same time to conduct the experiment without interfering with each other.
[0077] Experimental procedure: On the second day after modeling, a lead weight of 5% of the mouse's body weight was attached to the mouse's tail, and the mouse was placed in the aforementioned swimming pool. Timing was started from the moment the mouse entered the water, and the time it took to swim to exhaustion was recorded as the standard for exhaustion when the mouse's head was submerged in the water and it could not float to the surface within 5 seconds.
[0078] 4. Experimental Results
[0079] During the experiment, four animals died in the model control group (mortality rate 25%), and one animal died in the solvent control group (mortality rate approximately 6%). The remaining animals were in good condition and no deaths occurred. Specifically, the deaths in the model control group occurred before the rotator fatigue experiment (1 animal), before the exhaustive swimming experiment (1 animal), and after the exhaustive swimming experiment (2 animals). The deaths in the solvent control group occurred after the exhaustive swimming experiment (1 animal).
[0080] The animal weight measurement results are shown in Table 4. The measurement times were: before drug administration (on the day of pre-administration), before modeling (on the day of injection for modeling), and before tissue collection (on the day of dissection and tissue collection, i.e., the 3rd day after modeling). As can be seen from the results in Table 4, there was no significant difference in animal weight among the groups before modeling; compared with the blank control group, the animal weight in the model control group was significantly lower before tissue collection (P < 0.01), while there were no significant differences between the other groups and the model control group.
[0081] Table 4. Animal weight test results (g) Note: Compared with the blank control group, **P<0.01.
[0082] The results of the rotator fatigue test are shown in Table 5. As can be seen from the test results in Table 5, compared with the blank control group, the first drop time of animals in both the model control group and the solvent control group was significantly shortened (P < 0.05), and the number of drops was significantly increased (P < 0.01); compared with the model control group, there was no significant difference in the first drop time and the number of drops in the solvent control group; compared with the solvent control group, the first drop time of animals in the medium-dose Ganoderma lucidum spore oil group was significantly prolonged (P < 0.05), and the number of drops was significantly reduced (P < 0.05).
[0083] Table 5 Results of Rotor Fatigue Test Note: Compared with the blank control group, **P<0.01, *P<0.05; compared with the solvent control group, ## P < 0.01, # P < 0.05.
[0084] The results of the exhaustive swimming experiment are shown in Table 6. As can be seen from the test results in Table 6, compared with the blank control group, the exhaustive swimming time of animals in both the model control group and the solvent control group was significantly shortened (P < 0.01); compared with the model control group, there was no significant difference in the exhaustive swimming time of animals in the solvent control group; compared with the solvent control group, the exhaustive swimming time of animals in the medium-dose Ganoderma lucidum spore oil group was significantly prolonged (P < 0.05).
[0085] Table 6 Results of the Exhaustion Swimming Test Note: Compared with the blank control group, **P < 0.01; compared with the solvent control group, # P < 0.05.
[0086] Based on the detection data and differences between groups in Tables 4 to 6, the performance of the model in Example 2 within 3 days after modeling is analyzed as follows:
[0087] 1) Compared to the blank control group, the body weight of animals in each experimental group decreased to varying degrees after modeling. This decrease in body weight can reflect the fatigue status of the animals to some extent. 2) The results of the rotarod fatigue test and the exhaustive swimming test both showed that mice in the model control group and the solvent control group exhibited significant fatigue in various behavioral indicators at the test time points. Ganoderma lucidum spore oil significantly alleviated the fatigue state of the cancer-induced fatigue model mice, and various behavioral indicators could be restored to levels close to those of the blank control group, indicating that the cancer-induced fatigue model was successfully established. 3) Regarding the analysis of animal mortality, a large number of animals died in the model control group, while the mice in the other groups were generally in good condition. Only one animal died in the solvent control group. The underlying principle is speculated to be that compared to pre-administering sterile water (model control group), pre-administering a long-term oil-based solvent (solvent control group) increased the animals' tolerance to chemotherapy drugs, thereby reducing the mortality rate during the experiment and ensuring that the animals were in the most suitable state for drug evaluation without affecting the overall fatigue of the animals.
[0088] Example 2 only replaced the soybean oil used as the solvent in Example 1 with corn oil. The specific effects of several groups in the two example models are compared in Figures 1 and 2, and the analysis is as follows: In Example 2, one animal died in the solvent control group, while no animal deaths occurred in Example 1. Furthermore, the difference in exhaustion swimming time between the dosage groups of Ganoderma lucidum spore oil in Example 2 and the solvent control group was slightly less significant than in Example 1. Therefore, it can be seen that both corn oil and soybean oil can be used as oily solvents in this invention, but soybean oil in Example 1 is slightly more effective.
[0089] Comparative Example 1
[0090] Establishment of a fluorouracil-induced cancer-related fatigue model in mice and evaluation of therapeutic drugs (with short pre-dose administration time).
[0091] 1. Laboratory animals
[0092] C57BL / 6J mice, SPF grade, half male and half female, weighing 16-20g, a total of 96 mice, were housed in an SPF-grade animal room with free access to food, 12h light / 12h darkness, and an animal room temperature of 16-26℃ and relative humidity of 40%-70%.
[0093] 2. Experimental Materials
[0094] Modeling agent: Fluorouracil for injection.
[0095] Drug administration solvent: soybean oil.
[0096] Ganoderma lucidum spore oil: triglyceride content >90%, ergosterol content 0%–0.5%, and no Ganoderma lucidum triterpenoids detected. Prepare a solution of the appropriate concentration with soybean oil before administration; use immediately after preparation.
[0097] 3. Experimental Methods
[0098] (1) Establishment and administration of a fluorouracil-induced mouse model of cancer-related fatigue
[0099] Mice were randomly divided into 6 groups: blank control group, solvent control group, ultra-low dose group of Ganoderma lucidum spore oil, low dose group of Ganoderma lucidum spore oil, medium dose group of Ganoderma lucidum spore oil, and high dose group of Ganoderma lucidum spore oil, with 16 mice in each group, half male and half female. Animals in the blank control group and solvent control group were given solvent by gavage; animals in the ultra-low, low, medium, and high dose groups of Ganoderma lucidum spore oil were given Ganoderma lucidum spore oil by gavage at doses of 0.3, 0.6, 1.2, and 2.4 g / kg of daily, respectively. All groups were administered the drug once a day, with an administration volume of 5 mL / kg. The third day after modeling was selected as the sampling point, and the drugs were administered continuously once a day from 10 days before modeling until the day before sampling.
[0100] Ten days after pre-administration, animals in the blank control group were injected intraperitoneally with sterile water, while animals in the other groups were injected intraperitoneally with fluorouracil at a dose of 40 mg / kg once daily for 5 consecutive days. On the third day after modeling, the animals were dissected and their tissues were collected.
[0101] (2) Exhaustion Swimming Experiment
[0102] Experimental equipment and conditions: A constant temperature swimming pool was filled with water, with the water surface 5cm from the edge of the pool. The water temperature was set at 28℃ and the vortex was at its maximum. An acrylic sheet was used to divide the swimming pool into 4 areas. Four mice were placed in the 4 areas of the same swimming pool at the same time to conduct the experiment without interfering with each other.
[0103] Experimental procedure: On the second day after modeling, a lead weight of 5% of the mouse's body weight was attached to the mouse's tail, and the mouse was placed in the aforementioned swimming pool. Timing was started from the moment the mouse entered the water, and the time it took to swim to exhaustion was recorded as the standard for exhaustion when the mouse's head was submerged in the water and it could not float to the surface within 5 seconds.
[0104] 4. Experimental Results
[0105] Starting from day 1 after modeling, animals in all groups except the blank control group began to die consecutively. Details of animal mortality are shown in Table 7.
[0106] As shown in Table 7, the mortality rates of the animals within 3 days after modeling were as follows: approximately 19% in the solvent control group, 25% in the high-dose Ganoderma lucidum spore oil group, approximately 19% in the medium-dose group, approximately 13% in the low-dose group, and approximately 19% in the ultra-low-dose group. The cause of death is presumably attributed to organ failure caused by the side effects of chemotherapy drugs.
[0107] Table 7. Animal Mortality (number of animals)
[0108] The animal weight measurement results are shown in Table 8. The measurement times were: before drug administration (on the day of pre-drug administration), before modeling (on the day of injection for modeling), and before tissue collection (on the day of dissection and tissue collection, i.e., the 3rd day after modeling). As can be seen from the measurement results of each group in Table 8, there was no significant difference in animal weight among the groups before modeling; compared with the blank control group, the weight of animals in the solvent control group was significantly lower before tissue collection (P < 0.01).
[0109] Table 8. Animal weight test results (g) Note: Compared with the blank control group, **P<0.01.
[0110] The results of the exhaustive swimming experiment are shown in Table 9. As can be seen from the test results in Table 9, compared with the blank control group, the exhaustive swimming time of animals in the solvent control group was significantly shorter (P < 0.01); compared with the solvent control group, there was no significant difference in the exhaustive swimming time of animals in each dosage group of Ganoderma lucidum spore oil.
[0111] Table 9 Results of the Exhaustion Swimming Test Note: Compared with the blank control group, **P<0.01.
[0112] The results in Tables 7 and 9 indicate that in Comparative Example 1, with only 10 days of pre-administration, the various dosage groups of Ganoderma lucidum spore oil did not show efficacy on days 1 and 2 after modeling due to insufficient administration time, and the fatigue level of the animals did not decrease significantly. Furthermore, within 3 days after modeling, both the solvent control group and the various dosage groups of Ganoderma lucidum spore oil experienced continuous animal deaths, with the highest mortality rate reaching 25%. Therefore, Comparative Example 1 would struggle to obtain a sufficient amount of effective data at the final sampling stage, significantly increasing the cost required to successfully complete the experiment.
[0113] Compared with the model in Comparative Example 1, the pre-administration period in Example 1 was increased from 10 days to 15 days. The specific effects of several groups are shown in Figures 1 and 2, and the analysis is as follows: In Example 1, from day 1 to day 2 after modeling, the fatigue levels in each dosage group of Ganoderma lucidum spore oil were significantly lower than those in the solvent control group, indicating that the drug had already shown efficacy, and even recovered to levels close to those before modeling. Furthermore, no animal deaths occurred in either the solvent control group or the dosage groups of Ganoderma lucidum spore oil within 3 days after modeling. The differences between Example 1 and Comparative Example 1 show that, in terms of both modeling effect and drug evaluation effect, a pre-administration period of 15 days is superior to 10 days.
[0114] In summary, the method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs according to the present invention can effectively reflect the fatigue caused by the clinical application of these drugs, while significantly reducing animal mortality during the experiment. This is presumably due to the prolonged pre-administration of oil-based solvents, which to some extent improves the animals' tolerance to the chemotherapy drugs. Therefore, the model establishment method of the present invention has a high success rate, ensuring that the animals are in the most suitable state for drug evaluation without affecting their overall fatigue level. It can be used to screen drugs for the prevention and treatment of carcinogenic fatigue. Furthermore, because the animal mortality rate is significantly reduced and the observation period for animal behavior is shortened, high-quality experimental data can be obtained while effectively controlling experimental costs.
[0115] The above-disclosed embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention. Therefore, any equivalent variations made in accordance with the claims of the present invention are still within the scope of the present invention.
Claims
1. A method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs, characterized in that, The establishment method includes: administering oil-based solvents to experimental animals for 15-20 days prior to modeling.
2. The method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs according to claim 1, characterized in that, The oil-based solvent includes one or more of soybean oil, corn oil, olive oil, and sesame oil; the modeling process includes administering antimetabolite chemotherapy drugs to the experimental animals.
3. The method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs according to claim 1, characterized in that, The establishment method includes the following steps: A. The experimental animals were randomly divided into groups. The experimental animals in the blank control group were injected intraperitoneally with sterile water, while the experimental animals in the model control group, solvent control group, and positive drug group were injected intraperitoneally with antimetabolite chemotherapy drugs once a day for 5 consecutive days. The experimental animals in the blank control group and the model control group were given sterile water in advance before the intraperitoneal injection. The experimental animals in the solvent control group were given the oil-based solvent in advance before the intraperitoneal injection, and the experimental animals in the positive drug group were given a mixture of the oil-based solvent and the positive drug for cancer-induced fatigue in advance before the intraperitoneal injection. The advance administration time was 15-20 days, once a day. B. After 5 days of intraperitoneal injection, the animals in each group were assessed for fatigue using behavioral experiments.
4. The method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs according to claim 3, characterized in that, The pre-administration method was gavage; the pre-administration in step A started 15 to 20 days before the intraperitoneal injection of the experimental animals, once a day, and continued until the animal fatigue level was detected in step B.
5. The method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs according to claim 3, characterized in that, The experimental animals included mice; the daily dose of intraperitoneal injection for each group of experimental animals was 40 mg / kg b.wt.; the antimetabolite chemotherapy drug was fluorouracil; the volume of drugs administered to each group of experimental animals beforehand was 5 mL / kg b.wt.
6. The method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs according to claim 3, characterized in that, Step B includes: On days 1 to 3 after the completion of intraperitoneal injection for 5 days, behavioral experiments are used to detect the degree of fatigue in the experimental animals of each group; the behavioral experiments include one or more of the rotarod fatigue test and the exhaustive swimming test.
7. The method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs according to claim 6, characterized in that, The method for the rotator fatigue test includes: placing the experimental animal on the rotator, starting the timer from when the animal is placed on the rotator, and recording the time of the animal's first fall and the number of falls within 10 minutes.
8. The method for establishing a carcinogenic fatigue model induced by antimetabolite chemotherapy drugs according to claim 6, characterized in that, The method of the exhaustion swimming experiment includes: attaching a lead weight of 5% of the body weight to the tail of the experimental animal, placing the experimental animal in a swimming pool, starting the timer from the moment of entry into the water, and recording the exhaustion swimming time as the standard when the experimental animal's head sinks into the water and cannot float to the surface within 5 seconds.
9. A model established by a method for establishing a carcinogenic fatigue model of antimetabolite chemotherapeutic drugs according to any one of claims 1 to 8.
10. The application of the model according to claim 9 in studying cancer-related fatigue induced by antimetabolite chemotherapeutic agents and / or evaluating the efficacy of drug treatment for cancer-related fatigue.