CN-116868924-B - Method for establishing zebra fish cancer induced fatigue model

Abstract

The invention relates to the field of drug evaluation, and discloses a method for establishing a zebra fish cancer cause fatigue model, which aims to solve the problems that in an animal model of cancer cause fatigue in the prior art, the dosage of a chemotherapeutic drug is difficult to control, the accumulated toxicity of experimental animals is more and the experimental animals are easy to die, the modeling time is long, the flux is small and the operation is difficult. The model has short time for establishing, is simple to operate, and can efficiently and high-flux evaluate the anticancer effect of the medicine.

Inventors

• DAI MINGZHU
• WANG QINWEN
• Kong Jiexing
• LI CHUNQI

Assignees

• 杭州环特生物科技股份有限公司

Dates

Publication Date

20260512

Application Date

20221230

Claims (8)

1. 1. The application of the zebra fish cancer induced fatigue model in evaluating the anti-cancer induced fatigue effect of a drug is characterized in that the method for establishing the zebra fish cancer induced fatigue model comprises the following steps: A. Selecting normal-development zebra fish 4 days after fertilization, placing the zebra fish into a micro-pore plate, dividing the zebra fish into a normal control group and a model group, and performing intravenous injection of vinorelbine tartrate injection on the zebra fish in the model group, wherein the injection amount is 1-2 ng/tail, and the normal control group is not treated; B. culturing the normal control group and the model group in zebra fish culture solution at constant temperature; C. Comparing the total movement distance of the zebra fish in the control group with the total movement distance of the zebra fish in the model group, wherein the total movement distance of the model group is obviously smaller than that of the normal control group, so that the cancer-induced fatigue of the zebra fish is successfully established; D. After the zebra fish cancer induced fatigue is successfully established, randomly dividing a model group into a model control group and a drug group, and culturing the zebra fish in zebra fish culture solutions of a normal control group, the model control group and the drug group at 26-28 ℃ for 6-44 h, wherein the zebra fish culture solution of the drug group also comprises a drug to be detected; E. The total movement distance and ATP content of each group of zebra fish were detected and compared.
2. 2. The use according to claim 1, wherein in the model set of step a, anesthetic is added dropwise to the microplate prior to injection, reducing the difficulty of injection.
3. 3. The use according to claim 1, wherein in step B the zebra fish broth comprises 5mmol/L NaCl, 0.17mmol/L KCl, 0.4mmol/L CaCl 2 、0.16mmol/L MgSO 4 .
4. 4. Use according to any one of claims 1 to 3, wherein in step B the normal control group and the model group are incubated at 26-28 ℃.
5. 5. The use according to claim 4, wherein the incubation time of the normal control group and the model group in step B is 6-44 h.
6. 6. The use according to claim 1, wherein in step C the total movement distance of the zebra fish is determined using a behavioural analyzer.
7. 7. The use according to claim 6, wherein the behavior analyzer in step C sets the sensitivity Max parameter to 28-32.
8. 8. The use according to claim 1, wherein step C further comprises determining ATP content in the zebra fish of the normal control group and the model group.

Description

Method for establishing zebra fish cancer induced fatigue model Technical Field The invention relates to the field of drug evaluation, in particular to a method for establishing a zebra fish cancer induced fatigue model. Background Cancer-related fatigue (CRF) is one of the most common symptoms in cancer patients, and many reports indicate that 30% -60% of cancer patients experience moderate to severe fatigue during treatment, and even in some cases, cause discontinuation of cancer treatment. Cancer-induced fatigue is long in duration and has a great impact on work, mood and daily life, resulting in impaired overall quality of life during and after treatment. Symptoms of patients include debilitation, somnolence, bad mood, weakness, hypomnesis, etc., and cancer-induced fatigue cannot be relieved by rest. Numerous studies have been made at home and abroad on cancer-induced fatigue, but the physiological and pathological mechanisms and effective therapeutic means thereof are still not particularly clear, so that it is very important to build an animal model of cancer-induced fatigue for studying the mechanism of cancer-induced fatigue and screening related drugs. No standard modeling method has been available for animal models of cancer-induced fatigue. The common cancer-induced fatigue model is mainly a big mouse model. For example, sorensen et al selected 6-week male BALB/c mice as the molding subjects, established a cancer-induced fatigue model by intraperitoneally injecting oxaliplatin into the mice, loman et al molded by multiple injections of paclitaxel into the peritoneal cavity of 7-8-week female BALB/c mice. The mouse model needs to be injected with chemotherapeutics such as antibiotics, antimetabolites, alkaloids, alkylating agents, cytotoxins and the like for multiple times to complete the model, and because the mouse is small in size, the dose of the chemotherapeutics is difficult to control, and excessive accumulated toxicity is easy to cause to the mouse, so that the mouse dies. And the size and the mouse model have certain limitations in the research process, such as small flux, low speed, difficult operation and the like. Disclosure of Invention The invention provides a method for establishing a zebra fish cancer induced fatigue model, which aims to solve the problems that in the prior art, the dosage of a chemotherapeutic drug in an animal model for cancer induced fatigue is difficult to control, experimental animals accumulate more toxicity and are easy to die, the modeling time is long, the flux is small and the operation is difficult. In order to achieve the above purpose, the present invention adopts the following technical scheme: a method for establishing a zebra fish cancer induced fatigue model, comprising the following steps: A. Selecting normal-development zebra fish 4 days after fertilization, placing the zebra fish into a micro-pore plate, dividing the zebra fish into a normal control group and a model group, and performing intravenous injection of vinorelbine tartrate injection on the zebra fish in the model group, wherein the normal control group is not treated; B. culturing the normal control group and the model group in zebra fish culture solution at constant temperature; C. and comparing the total movement distance of the zebra fish in the control group with that of the model group, wherein the total movement distance of the model group is obviously smaller than that of the normal control group, so that the cancer-induced fatigue of the zebra fish is successfully established. According to the invention, zebra fish is used as a model organism, and vinorelbine tartrate is selected to induce the zebra fish to generate cancer induced fatigue. Vinorelbine tartrate is an antineoplastic agent that causes neurotoxic reactions, hematological toxicity during treatment, and thus causes fatigue in patients. In the invention, single injection molding is adopted in the molding process, so that the accumulated toxicity to the zebra fish is relatively small and controllable. The total movement distance of the zebra fish in the cultured model group is obviously reduced compared with that in the normal control group, and the model is successfully established when the reduction has statistical significance (p < 0.05). Preferably, in the model group of the step A, the injection quantity of the vinorelbine tartrate injection is 1-20 ng/tail. To reduce the difficulty of injection, anesthetic may be instilled into the microplate prior to injection. Preferably, in the step B, the zebra fish embryo culture solution comprises 5mmol/L NaCl, 0.17mmol/L KCl and 0.4mmol/LCaCl 2、0.16mmol/LMgSO4. Preferably, the normal control group and the model group in the step B are cultured at 26-28 ℃ for 6-44h. Preferably, in the step C, the total movement distance of the zebra fish is measured using a behavior analyzer. Preferably, the behavior analyzer in the step C sets the sensitivity Max parameter to 28-32. Prefe