CN114885900B - Method for constructing and evaluating in-vivo chemotherapeutic drug side effect research model - Google Patents
Method for constructing and evaluating in-vivo chemotherapeutic drug side effect research model Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
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Abstract
The invention discloses a construction method and an evaluation method of an in-vivo chemotherapeutic drug side effect research model, wherein the construction method comprises the following steps: preparing bromophenol blue dyed food, adding medicine, collecting Drosophila groups, placing into Drosophila tube, and administering to obtain a Drosophila animal model; the evaluation method comprises the following steps: detecting survival indexes, climbing indexes, abdomen scores after dyeing and antioxidation indexes of the model group and the normal control group. The invention uses the dye food containing the chemotherapeutics to feed the drosophila to construct an animal model for researching the side effects of the chemotherapeutics in vivo, uses the normal food as a control, counts the survival rate of the drosophila, and carries out exercise capacity measurement, antioxidant stress capacity measurement and ingestion amount measurement when drosophila starts to die after feeding the food containing cytarabine/irinotecan hydrochloride.
Description
Technical Field
The invention belongs to the technical field of animal model evaluation, and particularly relates to a method for constructing and evaluating an in-vivo chemotherapeutic drug side effect research model.
Background
According to the statistics of world health organization, the global cancer death cases in 2020 are 996 ten thousand, which account for about 50% of new cases, and the incidence and death rate of global cancers are in an ascending trend. The treatment regimen adopted by patients at different stages of disease progression is different, and chemotherapy is often dominant among them. However, chemotherapeutic drugs have general cell killing properties, which cause certain damage to normal cells while killing cancer cells, resulting in cardiotoxicity, lung injury, nephrotoxicity, hepatotoxicity, gastrointestinal toxicity, neurotoxicity, etc., affecting the course of chemotherapy in tumor patients, and serious or even life-threatening.
The current research on the side effects of chemotherapeutic drugs is mostly in vivo or in vitro cell experiments, and in vivo experiments, the model building time of the clock mammal is long, the cost is high, and animal ethics and other factors exist, so that the research is limited. In vitro experiments only make toxicity assessment at the level of single cells in vitro, but the tested animals are much more complex than single cells, the metabolism, immune and other systems are mutually connected, even the test of single poison is subject to unpredictable interference, the verification of the metabolism dynamics at the level of biological integrity is not carried out, and the overall toxicity after the action of each system in vivo environment cannot be accurately estimated. Therefore, a more suitable animal model for side effects of the chemotherapeutic drugs is lacking, so that the research and screening of the mechanism of the side effects of the chemotherapeutic drugs can relieve the bottleneck of the side effects of the chemotherapeutic drugs.
Drosophila (Drosophila melanogaster) is an ideal model organism for researching the occurrence mechanism of human diseases, and along with the continuous enrichment of a drosophila pathology model and a transgenic drosophila strain, the drosophila model is widely applied to the research of the disease mechanisms of reproductive, immune, nervous, cardiovascular and other systems, neurodegenerative diseases, metabolic syndrome, sleep and the like. Drosophila has a digestive system and a metabolic system similar to those of mammals, is widely applied to drug toxicity evaluation, and has potential application value in researching side effects of chemotherapeutic drugs.
Disclosure of Invention
The invention aims at providing a method for constructing an in vivo chemotherapeutic drug side effect research model;
the second purpose of the invention is to provide an evaluation method of an in vivo chemotherapeutic drug side effect research model;
the animal model is constructed and evaluated to establish a high throughput screening platform for alleviating the lineages of drugs that cause damage to the body by chemotherapeutic drugs.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a construction method of an in vivo chemotherapeutic drug side effect research model comprises the following steps:
s1: adding bromophenol blue dye into Drosophila food, uniformly mixing to obtain dyed food, equally dividing the dyed food, filling the dyed food into a Drosophila tube, respectively placing prepared cytarabine and irinotecan hydrochloride into the Drosophila tube, and marking to obtain cytarabine Drosophila tube and irinotecan hydrochloride Kang Guoying tube;
s2: CO from Drosophila 2 After anesthesia, 3-day-old male and female drosophila was collected and randomly grouped20 tubes are separated from each other, each group is repeated 3 times, and the two tubes are respectively placed in cytarabine drosophila tube and irinotecan hydrochloride drosophila tube obtained in S1;
s3: updating stained food every 2-3 days, and administering for 9-10 days to obtain Drosophila animal model.
In order to further realize the invention, the concentration of cytarabine in S1 is 1mM-10mM, and the concentration of the hydrochloric acid of the yi Li Tikang is 1mM-5mM.
In order to further realize the invention, the concentration of the bromophenol blue dye in S1 is 0.01% -0.5%.
The evaluation method of the construction method of the in vivo chemotherapeutic drug side effect research model is that a normal control group is established, the Drosophila animal model obtained in the step S3 is used as a model group, and the following indexes of the detection model group and the normal control group are as follows:
(1) Survival indexes;
(2) Climbing indexes;
(3) Scoring abdomen after staining;
(4) An antioxidant index;
and carrying out differential analysis on the index data and the scores of the model group and the normal control group by using a statistical method, and setting up a qualified value for the index, and if the index data and the scores of the model group have significant differences compared with the normal control group and the model group reaches the qualified value, considering that the model is successfully constructed.
In order to further realize the invention, the method for detecting the survival index comprises the following steps: continuing to administer the drosophila animal model obtained in the step S3, observing the condition of the drosophila 3-4 times a day, recording the death condition of the drosophila until all drosophila in the model group with the cytarabine administration concentration being more than or equal to 5mM or the cytarabine hydrochloride administration concentration being more than or equal to 2.5mM die, and calculating the survival rate of the model group:
compared with the survival rate of a normal control group, the model group has obvious difference, the survival rate of the cytarabine administration model group on the 22 th day is lower than 70 percent, and the model group is qualified, and the survival rate of the irinotecan hydrochloride administration model group on the 22 th day is lower than 80 percent.
In order to further realize the invention, the detection method of the climbing index comprises the following steps: transferring the Drosophila animal model obtained in the step S3 and the normal control group into a climbing pipe with the length of 15cm, recording the number of Drosophila climbing 8cm for 8S in the climbing pipe, and calculating the climbing index of the model group:
compared with the survival rate of the normal control group, the model group has obvious difference, and the model group climbing index is lower than 65 percent, namely the model group climbing index is qualified.
In order to further realize the invention, the method for detecting the abdomen score after dyeing comprises the following steps: transferring the Drosophila animal model obtained in S3 and normal control group into Drosophila tube containing 1% agar for starvation for 16-21 hr, transferring into Drosophila tube containing 2% bromophenol blue dye, feeding for 3-8 hr, and collecting Drosophila CO 2 After anesthesia at CO 2 Abdomen scoring for each group of drosophila on the ventilation plate:
(1) Food with blue dye occupies less than one third of the abdomen volume and is beaten by 1 minute;
(2) Blue dye food accounts for 2 minutes of the abdomen of Drosophila;
(3) The blue dye content exceeds half of the abdomen for 3 minutes;
and (5) the abdomen score after the study model group is dyed is lower than 1.75, and the study model group is qualified.
In order to further realize the invention, the detection method of the antioxidant index comprises the following steps: transferring the drosophila animal model obtained in the step S3 and the normal control group into an empty drosophila tube for starving for 1h, transferring into the drosophila tube paved with filter paper, dripping 300 mu L of hydrogen peroxide solution, transferring the starved drosophila into the tube after the hydrogen peroxide solution completely permeates the filter paper, observing the condition of the drosophila 3-4 times per day, changing agar every 2-3 days until all the drosophila in the study model group and the normal control group die, and calculating the survival rate of the drosophila:
the survival rate of the male drosophila in 42h in the research model group is lower than 30 percent, and the survival rate of the female drosophila in 42h is lower than 70 percent.
A Drosophila tube for constructing an in vivo chemotherapy drug side effect research model comprises a tube body and a cover body, wherein a dyed food layer is arranged at the bottom of the tube body, a tray is arranged on the top surface of the dyed food layer, and a drug administration layer is arranged in the tray.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, drosophila food containing a chemotherapeutic drug cytarabine or irinotecan hydrochloride is fed to drosophila at 3 days old so as to construct an animal (drosophila) model for researching side effects of the chemotherapeutic drug in vivo, normal food is used as a control, survival rate of the drosophila is counted, and after the cytarabine/irinotecan hydrochloride-containing food is fed, the drosophila begins to die, the movement capacity measurement, the antioxidant stress capacity measurement and the food intake measurement are carried out, and the result shows that the cytarabine/irinotecan hydrochloride Li Tikang causes the increase of the lethality, the movement capacity, the stress resistance capacity and the food intake of the drosophila, so that whether the model is constructed successfully or not is evaluated. Compared with other traditional model organisms, the method has the advantages of high breeding speed, low breeding cost, short experimental period and remarkable experimental effect of the drosophila melanogaster in the experimental period of 25 days, and can be used for establishing and evaluating a damaged animal (drosophila melanogaster) model with side effects caused by chemotherapeutic drugs in the drosophila melanogaster body, thereby establishing a high-throughput screening platform for relieving the lineages of the drugs for reducing the damage of the body caused by the chemotherapeutic drugs.
The invention improves the traditional administration mode, the traditional administration mode is to dissolve the medicine in basic food, the medicine is poured into the bottom of the drosophila tube, drosophila can not independently select to eat and can only eat forcedly, and the drosophila can meet the daily consumption only by needing less food, and the drosophila tube has the phenomena of larger area, reagent waste and the like to a certain extent, so that the experimental cost is higher. In contrast, the novel drosophila tube is designed for administration, and the non-metabolizable bromophenol blue dye is added into food, so that whether the drosophila belly has a blue area or not can be observed in real time to ensure that the drosophila ingests food containing medicines, meanwhile, the drosophila can independently select drinking water or eat food because the periphery of the tray is 1% of agar, the autonomous diet of the drosophila is greatly increased, and the autonomous diet selectivity similar to that of mammals is simulated as much as possible.
The invention also supplements the existing model organism for researching the side effect of the chemotherapeutic drug, makes up the defect of lack of integrity of in vitro cell experiments, enriches the variety of in vivo experimental model animals, and greatly reduces research economy and time cost. Is very favorable for developing the research of the pathogenesis of the side effect of the chemotherapeutic drugs and the high-throughput screening of the anti-chemotherapeutic drugs.
Drawings
Fig. 1 shows survival curves of Drosophila fed with cytarabine at different concentrations, expressing the effect of cytarabine at different concentrations on the life span of Drosophila, wherein fig. A shows survival curves of male flies fed with cytarabine at different concentrations (n=100-120), and fig. B shows survival curves of female flies fed with cytarabine at different concentrations (n=100-120). * P <0.0001 indicates that the difference is statistically significant;
fig. 2 shows survival curves of flies raised with irinotecan hydrochloride at different concentrations in the present invention, expressing the effect of irinotecan hydrochloride at different concentrations on the life span of drosophila, wherein a is a survival curve (n=74-120) of flies raised with irinotecan hydrochloride at different concentrations, B is a survival curve (n=73-120) of flies raised with irinotecan hydrochloride at different concentrations, p <0.01, p <0.0001 indicates that the difference is statistically significant;
fig. 3 shows climbing indexes of cytarabine with different concentrations for feeding drosophila, and expresses the influence of cytarabine with different concentrations on the exercise capacity of drosophila, wherein a graph a shows climbing indexes (n=8) of 8cm climbed by 8s of male flies fed with cytarabine with different concentrations for 10 days, and a graph B shows climbing indexes (n=8) of 8cm climbed by 8s of female flies fed with cytarabine with different concentrations for 10 days, wherein p <0.01 and p <0.0001 represent that differences are statistically significant;
fig. 4 shows belly scores and absorbance of drosophila fed with cytarabine at different concentrations, expressing the effect of cytarabine at different concentrations on drosophila feed intake, wherein a graph a shows belly scores (n=79) of male flies fed with cytarabine at different concentrations for 10 days, and a graph B shows belly scores (n=90) of female flies fed with cytarabine at different concentrations for 10 days;
fig. 5 shows survival curves of male flies raised for 10 days under oxidative stress conditions, in which the influence of cytarabine at different concentrations on the antioxidant stress capability of drosophila is expressed, a graph a shows survival curves of male flies raised for 10 days under oxidative stress conditions (n=160), and a graph B shows survival curves of female flies raised for 10 days under oxidative stress conditions (n=140), wherein p <0.001, p <0.0001 indicate that the difference is statistically significant;
FIG. 6 is a schematic view of the structure of Drosophila tube in the present invention;
the reference numerals have the following meanings: 1. a tube body; 2. a cover body; 3. staining the food layer; 4. a tray; 5. and a drug administration layer.
Detailed Description
The invention is further described below with reference to the drawings and the detailed description.
A construction method of an in vivo chemotherapeutic drug side effect research model comprises the following steps:
s1: adding bromophenol blue dye into Drosophila food, uniformly mixing to obtain dyed food, equally dividing the dyed food, filling the dyed food into a Drosophila tube, respectively placing prepared cytarabine and irinotecan hydrochloride into the Drosophila tube, and marking to obtain cytarabine Drosophila tube and irinotecan hydrochloride Kang Guoying tube;
s2: CO from Drosophila 2 After anesthesia, collecting 3-day-old male and female drosophila, randomly grouping, separating 20 male and female drosophila from each other, repeating each group for 3 times, and respectively placing in the cytarabine drosophila tube and irinotecan hydrochloride drosophila tube obtained in the step S1;
s3: updating stained food every 2-3 days, and administering for 9-10 days to obtain Drosophila animal model.
The concentration of cytarabine in S1 is 1mM-10mM, the concentration of the hydrochloric acid I Li Tikang is 1mM-5mM, and the concentration of the bromophenol blue dye is 0.01% -0.5%.
The method for constructing the in vivo chemotherapeutic drug side effect research model in embodiment 1 comprises the following steps:
s1: adding bromophenol blue dye into Drosophila food, uniformly mixing to obtain dyed food, equally dividing the dyed food, filling the dyed food into Drosophila tubes, preparing cytarabine-containing hydrochloric acid Li Tikang with the concentration of 1mM, 5mM and 10mM and irinotecan hydrochloride with the concentration of 1mM, 2.5mM and 5mM respectively, respectively placing the prepared cytarabine and irinotecan hydrochloride into the Drosophila tubes, and marking to obtain cytarabine Drosophila tubes with low administration group, medium administration group and high administration group and irinotecan Kang Guoying tubes with low administration group, medium administration group and high administration group;
s2: CO from Drosophila 2 After anesthesia, collecting 3-day-old male and female drosophila, randomly grouping, separating 20 male and female drosophila from each other, repeating each group for 3 times, and respectively placing in the cytarabine drosophila tube and irinotecan hydrochloride drosophila tube obtained in the step S1;
s3: updating stained food every 2-3 days, and administering for 9-10 days to obtain Drosophila animal model.
Example 2, chemotherapeutic drugs have general killing power to organisms, viability is one of important indexes reflecting whether the drugs have toxicity, and the toxicity of the chemotherapeutic drugs to organisms can be intuitively illustrated by detecting the survival indexes of drosophila fed with the chemotherapeutic drugs. The detection method of the survival index comprises the following steps:
continuing to administer the drosophila animal model obtained in the step S3, observing the condition of the drosophila 3-4 times a day, recording the death condition of the drosophila until all drosophila in the model group with the cytarabine administration concentration being more than or equal to 5mM or the cytarabine hydrochloride administration concentration being more than or equal to 2.5mM die, and calculating the survival rate of the model group:
compared with the survival rate of a normal control group, the model group has obvious difference, the survival rate of the cytarabine administration model group on the 22 th day is lower than 70 percent, and the model group is qualified, and the survival rate of the irinotecan hydrochloride administration model group on the 22 th day is lower than 80 percent.
As can be seen from fig. 1-2, cytarabine and irinotecan hydrochloride can significantly shorten the life span of drosophila, and as the concentration of chemotherapeutic agent increases, the life span of drosophila gradually shortens. Studies have shown that chemotherapeutic agents have a lethal effect. Wherein the survival rate of male flies is 40.09% when the cytarabine is fed for 22 days, the survival rate of female flies is 65.50%, the survival rate of male flies is 0 when the cytarabine is fed for 22 days, the survival rate of female flies is 1.65%, and the survival rates of male flies and female flies are 0 when the cytarabine is fed for 22 days and 10 mM. The survival rate of the male flies is 10.42% when the 1mM irinotecan hydrochloride is fed for 22 days, the survival rate of the female flies is 78.28%, the survival rate of the male flies is 0 when the 2.5mM irinotecan hydrochloride is fed for 22 days, the survival rate of the female flies is 32.61%, and the survival rates of the male flies and the female flies are 0 when the 5mM irinotecan hydrochloride is fed for 22 days.
From the above, the survival rates of cytarabine low administration group, medium administration group and high administration group and irinotecan hydrochloride low administration group, medium administration group and high administration group have significant difference compared with the survival rate of the normal control group, and the survival index detection is qualified, namely the model construction is successful.
Embodiment 3, climbing is the embodiment of the exercise capacity of the drosophila, the nervous system of the drosophila can regulate and control the exercise capacity of the drosophila, and when the nerve injury of the organism is caused by the chemotherapy medicine, the exercise capacity of the drosophila can be reduced. The strength of the fruit fly movement capacity can be reflected by detecting the fruit fly climbing index. The detection method of the climbing index comprises the following steps:
transferring the Drosophila animal model obtained in the step S3 and the normal control group into a climbing pipe with the length of 15cm, recording the number of Drosophila climbing 8cm for 8S in the climbing pipe, and calculating the climbing index of the model group:
compared with the survival rate of the normal control group, the model group has obvious difference, and the model group climbing index is lower than 65 percent, namely the model group climbing index is qualified.
As can be seen from fig. 3, cytarabine can reduce the exercise ability of drosophila, and the climbing ability of drosophila gradually decreases with increasing concentration. Studies have shown that cytarabine can cause nerve damage, resulting in reduced motor ability. Wherein, the climbing index of the male flies in the normal control group is 91.23%, and the climbing indexes of the male flies in the cytarabine low administration group, the cytarabine medium administration group and the cytarabine high administration group are 64.12%, 29.17% and 16.38%, respectively, which are 70.28%, 31.98% and 17.95% of the normal control group; the climbing index of the female flies in the normal control group is 83.74%, and the climbing indexes of the female flies in the cytarabine low-dosing group, the cytarabine medium-dosing group and the cytarabine high-dosing group are 72.62% (without statistical significance), 40.13% and 18.88%, respectively 86.73%, 47.92% and 22.55% of the normal control group
From the above, the climbing indexes of the cytarabine low administration group, the cytarabine medium administration group and the cytarabine high administration group have a significant difference compared with the climbing index of the normal control group, and the climbing indexes are detected to be qualified, namely the model is successfully constructed.
Example 4, feeding amount is one of the indexes of normal physiology of the organism, and when the chemotherapy drugs cause injury of the organism, the feeding of the drosophila is affected, and the feeding condition can be reflected by detecting the abdominal score and absorbance of the drosophila. The detection method of the abdomen score after staining comprises the following steps:
transferring the Drosophila animal model obtained in S3 and normal control group into Drosophila tube containing 1% agar for starvation for 16-21 hr, transferring into Drosophila tube containing 2% bromophenol blue dye, feeding for 3-8 hr, and collecting Drosophila CO 2 After anesthesia at CO 2 Abdomen scoring for each group of drosophila on the ventilation plate:
(1) Food with blue dye occupies less than one third of the abdomen volume and is beaten by 1 minute;
(2) Blue dye food accounts for 2 minutes of the abdomen of Drosophila;
(3) The blue dye content exceeds half of the abdomen for 3 minutes;
and (5) the abdomen score after the study model group is dyed is lower than 1.75, and the study model group is qualified.
As can be seen from fig. 4, cytarabine can reduce the feeding amount of drosophila, and the feeding amount of drosophila gradually decreases with the increase of concentration. Studies have shown that cytarabine can cause metabolic disorders, reducing food intake. The abdomen score of the male flies in the normal control group is 2.07 points, and the abdomen scores of the male flies in the cytarabine low administration group, the cytarabine medium administration group and the cytarabine high administration group are 1.88 points (without statistical significance), 1.76 points (without statistical significance) and 1.39 points, which are 90.82 percent, 85.02 percent and 67.15 percent of the normal control group respectively; the normal control group female abdomen score was 2.03 points, and the cytarabine low, medium and high groups female abdomen scores were 1.54 points, 1.64 points and 1.52 points, respectively 75.86%, 80.79% and 78.88% of the normal control group.
From the above, the stained abdominal scores of cytarabine low administration group, medium administration group and high administration group have significant difference compared with the stained abdominal score of the normal control group, and the detection of the stained abdominal score is qualified, namely the model construction is successful.
Example 5 under normal conditions, the body has very strong anti-oxidative stress ability, and when the body is damaged, the anti-oxidative stress ability of the body is reduced, thereby causing damage to the body. The damage condition of the medicine to the organism can be reflected by detecting the antioxidant index of the drosophila. The detection method of the antioxidant stress index comprises the following steps: transferring the drosophila animal model obtained in the step S3 and the normal control group into an empty drosophila tube for starving for 1h, transferring into the drosophila tube paved with filter paper, dripping 300 mu L of hydrogen peroxide solution, transferring the starved drosophila into the tube after the hydrogen peroxide solution completely permeates the filter paper, observing the condition of the drosophila 3-4 times per day, changing agar every 2-3 days until all the drosophila in the study model group and the normal control group die, and calculating the survival rate of the drosophila:
the survival rate of the male drosophila in 42h in the research model group is lower than 30 percent, and the survival rate of the female drosophila in 42h is lower than 70 percent.
As can be seen from FIG. 5, cytarabine can reduce the antioxidant stress capacity of Drosophila, indicating that cytarabine can cause oxidative damage to the body. The survival rate of the normal control group male flies is 42.17% when the hydrogen peroxide is stimulated for 42 hours, and the survival rates of the male flies in the low-administration group, the middle-administration group and the high-administration group of cytarabine are 41.31% (without statistical significance), 38.96% (without statistical significance) and 27.96% respectively; the survival rate of the normal control female flies was 87.62%, and the survival rates of the female flies in the low-dose group, the medium-dose group and the high-dose group of cytarabine were 88.76% (without statistical significance), 65.16% and 63.99%, respectively.
From the above, the survival rates of cytarabine low administration group, cytarabine medium administration group and cytarabine high administration group are obviously different from that of the normal control group, and the detection of the anti-oxidative stress index is qualified, namely the model is successfully constructed.
As shown in fig. 6, a drosophila tube constructed by an in vivo chemotherapeutic drug side effect research model, wherein the drosophila tube in S2 comprises a tube body 1 and a cover body 2, a dyed food layer 3 is arranged at the bottom of the tube body 1, a tray 4 is arranged on the top surface of the dyed food layer 3, and a drug administration layer 5 is arranged in the tray 4.
Claims (9)
1. The method for constructing the research model of the side effect of the chemotherapeutic drug in vivo is characterized by comprising the following steps:
s1: adding bromophenol blue dye into Drosophila food, uniformly mixing to obtain dyed food, equally dividing the dyed food, filling the dyed food into a Drosophila tube, respectively placing prepared cytarabine and irinotecan hydrochloride into the Drosophila tube, and marking to obtain cytarabine Drosophila tube and irinotecan hydrochloride Kang Guoying tube;
s2: CO from Drosophila 2 After anesthesia, collecting 3-day-old male and female drosophila, randomly grouping, separating 20 male and female drosophila from each other, repeating each group for 3 times, and respectively placing in the cytarabine drosophila tube and irinotecan hydrochloride drosophila tube obtained in the step S1;
s3: updating stained food every 2-3 days, and administering for 9-10 days to obtain Drosophila animal model.
2. The method for constructing an in vivo chemotherapeutic drug side effect research model according to claim 1, which is characterized in that: the concentration of cytarabine in S1 is 1mM-10mM, and the concentration of the cytarabine in the hydrochloric acid is 1mM-5mM.
3. The method for constructing an in vivo chemotherapeutic drug side effect research model according to claim 1, which is characterized in that: the concentration of the bromophenol blue dye in S1 is 0.01% -0.5%.
4. The method for evaluating a method for constructing an in vivo chemotherapeutic drug side effect research model according to any one of claims 1 to 3, wherein a normal control group is established, the drosophila animal model obtained in S3 is used as a model group, and the following indexes of the model group and the normal control group are detected:
(1) Survival indexes;
(2) Climbing indexes;
(3) Scoring abdomen after staining;
(4) An antioxidant index;
and carrying out differential analysis on the index data and the scores of the model group and the normal control group by using a statistical method, and setting up a qualified value for the index, and if the index data and the scores of the model group have significant differences compared with the normal control group and the model group reaches the qualified value, considering that the model is successfully constructed.
5. The method for evaluating the method for constructing an in vivo chemotherapeutic drug side effect study model according to claim 4, wherein: the detection method of the survival index comprises the following steps: continuing to administer the drosophila animal model obtained in the step S3, observing the condition of the drosophila 3-4 times a day, recording the death condition of the drosophila until all drosophila in the model group with the cytarabine administration concentration being more than or equal to 5mM or the cytarabine hydrochloride administration concentration being more than or equal to 2.5mM die, and calculating the survival rate of the model group:
compared with the survival rate of a normal control group, the model group has obvious difference, the survival rate of the cytarabine administration model group on the 22 th day is lower than 70 percent, and the model group is qualified, and the survival rate of the irinotecan hydrochloride administration model group on the 22 th day is lower than 80 percent.
6. The method for evaluating the method for constructing an in vivo chemotherapeutic drug side effect study model according to claim 5, wherein: the detection method of the climbing index comprises the following steps: transferring the Drosophila animal model obtained in the step S3 and the normal control group into a climbing pipe with the length of 15cm, recording the number of Drosophila climbing 8cm for 8S in the climbing pipe, and calculating the climbing index of the model group:
compared with the survival rate of the normal control group, the model group has obvious difference, and the model group climbing index is lower than 65 percent, namely the model group climbing index is qualified.
7. The method for evaluating the method for constructing an in vivo chemotherapeutic drug side effect study model according to claim 6, wherein: the detection method of the abdomen score after dyeing comprises the following steps: transferring the Drosophila animal model obtained in S3 and normal control group into Drosophila tube containing 1% agar for starvation for 16-21 hr, transferring into Drosophila tube containing 2% bromophenol blue dye, feeding for 3-8 hr, and collecting Drosophila CO 2 After anesthesia at CO 2 Abdomen scoring for each group of drosophila on the ventilation plate:
(1) Food with blue dye occupies less than one third of the abdomen volume and is beaten by 1 minute;
(2) Blue dye food accounts for 2 minutes of the abdomen of Drosophila;
(3) The blue dye content exceeds half of the abdomen for 3 minutes;
and (5) the abdomen score after the study model group is dyed is lower than 1.75, and the study model group is qualified.
8. The method for evaluating the method for constructing an in vivo chemotherapeutic drug side effect study model according to claim 7, wherein: the detection method of the antioxidant index comprises the following steps: transferring the drosophila animal model obtained in the step S3 and the normal control group into an empty drosophila tube for starving for 1h, transferring into the drosophila tube paved with filter paper, dripping 300 mu L of hydrogen peroxide solution, transferring the starved drosophila into the tube after the hydrogen peroxide solution completely permeates the filter paper, observing the condition of the drosophila 3-4 times per day, changing agar every 2-3 days until all the drosophila in the study model group and the normal control group die, and calculating the survival rate of the drosophila:
the survival rate of the male drosophila in 42h in the research model group is lower than 30 percent, and the survival rate of the female drosophila in 42h is lower than 70 percent.
9. A drosophila catheter for use in a method of constructing a model for studying side effects of chemotherapeutic agents in vivo as defined in claim 1, wherein: the drosophila tube comprises a tube body (1) and a cover body (2), wherein a dyed food layer (3) is arranged at the bottom of the tube body (1), a tray (4) is arranged on the top surface of the dyed food layer (3), and a drug administration layer (5) is arranged in the tray (4).
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| US6291739B1 (en) * | 2000-03-24 | 2001-09-18 | Council Of Scientific And Industrial Research | Method for screening of potential anti-epileptic drugs using a Drosophila melanogaster model |
| CN109620828A (en) * | 2018-12-19 | 2019-04-16 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Method for establishing model, model and the application of chemotherapy metenteron adverse reaction |
| CN113100178A (en) * | 2021-04-07 | 2021-07-13 | 华北理工大学 | Method for establishing drosophila melanogaster tumor invasion model |
| CN113637702A (en) * | 2021-07-23 | 2021-11-12 | 珠海市人民医院 | Construction method and application of Drosophila model of Alzheimer's disease |
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| US6291739B1 (en) * | 2000-03-24 | 2001-09-18 | Council Of Scientific And Industrial Research | Method for screening of potential anti-epileptic drugs using a Drosophila melanogaster model |
| CN109620828A (en) * | 2018-12-19 | 2019-04-16 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Method for establishing model, model and the application of chemotherapy metenteron adverse reaction |
| CN113100178A (en) * | 2021-04-07 | 2021-07-13 | 华北理工大学 | Method for establishing drosophila melanogaster tumor invasion model |
| CN113637702A (en) * | 2021-07-23 | 2021-11-12 | 珠海市人民医院 | Construction method and application of Drosophila model of Alzheimer's disease |
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