CN115104575A - Establishment method and application of alcoholic depression animal model - Google Patents
Establishment method and application of alcoholic depression animal model Download PDFInfo
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- CN115104575A CN115104575A CN202210805716.3A CN202210805716A CN115104575A CN 115104575 A CN115104575 A CN 115104575A CN 202210805716 A CN202210805716 A CN 202210805716A CN 115104575 A CN115104575 A CN 115104575A
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- 238000010171 animal model Methods 0.000 title claims abstract description 51
- 230000001476 alcoholic effect Effects 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 29
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 52
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- 230000001154 acute effect Effects 0.000 claims abstract description 30
- 230000006698 induction Effects 0.000 claims abstract description 29
- 230000007267 depressive like behavior Effects 0.000 claims abstract description 13
- 230000002075 anti-alcohol Effects 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- 229940079593 drug Drugs 0.000 claims description 6
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- 230000001939 inductive effect Effects 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
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- 238000002474 experimental method Methods 0.000 description 12
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- 208000007848 Alcoholism Diseases 0.000 description 10
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/02—Breeding vertebrates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- 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
Abstract
The invention discloses a method for establishing an alcoholic depression animal model, which comprises the following steps: selecting experimental animals meeting experimental requirements, and pretreating the experimental animals subjected to multiple times of intragastric administration with acute alcohol with preset intragastric administration amount; after the acute alcohol pretreatment, LPS is adopted to induce the experimental animal until a depression-like behavior appears, and the constructed alcoholic depression model is verified; the preset gavage amount is measured by gavage of each experimental animal by 5-10g/kg according to the weight of the experimental animal, preferably induced by a mode of gavage LPS (low-temperature stress) and better accords with the actual situation of alcoholic depression, and is measured by gavage of each experimental animal by 10-15mg/kg according to the weight of the experimental animal; by adopting the method, the depression behavior can be induced within 2-4 days, compared with the chronic alcohol induction period, the method greatly shortens the experimental modeling time, the depression behavior of the mice is obvious, and the constructed alcoholic depression model has good effect.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for establishing an alcoholic depression animal model and application thereof.
Background
Depression is a common disorder of mental activities, and its typical symptoms include depressed mood, thought retardation, decreased interest, eating and sleeping disorders, cognitive impairment, and other somatic symptoms. In recent years, the incidence rate of depression is continuously increased, at least 80 ten thousand of patients with depression die by suicide every year, the incidence rate of depression is 12% -17% in the life of the people, and alcoholic depression easily forms a vicious circle to further aggravate the symptoms of depression.
Although the situation is so severe, the etiology and pathogenesis of alcoholic depression are not completely understood so far, most of the existing depression models are different from the actual situation of alcoholic depression, and different alcoholic depression animal models are constructed in a targeted manner for comprehensively understanding the pathogenesis of alcoholic depression or developing anti-alcoholic depression drugs. In the research of the relation between alcohol and depression, chronic alcohol treatment can induce depression-like behavior, while acute alcohol treatment alone can hardly induce depression behavior, but the chronic alcohol induction period is long, and at present, no animal model can completely cover all symptoms of depression.
Therefore, the research on a rapid induction alcoholic depression model is beneficial to comprehensively understanding the pathogenesis of alcoholic depression and is beneficial to promoting the research on potential mechanisms of relation between alcohol and depression or anti-alcoholic depression drugs.
Disclosure of Invention
Aiming at the defects or the improvement requirements of the prior art, the invention provides a method for establishing an alcoholic depression animal model and application thereof, and aims to adopt combination of alcohol gavage and LPS to induce the alcoholic depression model, thereby solving the technical problems of long induction period and single model type of the existing alcoholic depression model.
To achieve the above object, according to one aspect of the present invention, there is provided a method for establishing an animal model of alcoholic depression, comprising the steps of:
selecting experimental animals meeting experimental requirements;
pretreating an experimental animal subjected to multiple times of intragastric administration with acute alcohol with a preset intragastric administration amount; the preset gavage amount is calculated by 5-10g/kg of gavage of each experimental animal according to the weight of the experimental animal;
and (3) after acute alcohol pretreatment, adopting LPS to induce the experimental animal until the experimental animal has depression-like behavior, and verifying the constructed alcoholic depression model.
Preferably, the LPS induction of the animal model of alcoholic depression is induced by means of intragastric administration of LPS, and the weight of each animal is 10-15mg/kg per intragastric administration according to the weight of the animal.
Preferably, the multiple times of gavage are gavage for 3-5 times, and the interval time of each time is 12-18 h.
Preferably, in the method for establishing the animal model of alcoholic depression, LPS is adopted to induce the experimental animal 2-6 hours after the acute alcohol gavage pretreatment.
Preferably, the acute alcohol of the animal model for alcoholic depression is perfused for 3 times at an interval of 12h according to the body weight of the experimental animal and calculated by 5g/kg per each perfusion.
Preferably, the LPS induction of the animal model for alcoholic depression is single induction, and the induction is carried out according to the body weight of an experimental animal and the amount of each gavage is 10 mg/kg.
According to another aspect of the invention, a depression model constructed by the alcoholic depression model building method is provided.
According to another aspect of the invention, the application of the depression model obtained by the alcoholic depression model building method in screening anti-alcoholic depression drugs is also provided.
In general, compared with the prior art, the technical scheme of the invention has the following beneficial effects due to the combined induction of acute alcohol and LPS:
according to the weight of the experimental animal, each experimental animal is gavaged with 5-10g/kg of acute alcohol every time, the experimental animal is firstly subjected to repeated gavage treatment by adopting acute alcohol, then LPS (low-pressure lipoprotein) is adopted for induction, an alcoholic depression model can be induced within 2-4 days, the depression behavior of a mouse is obvious, and the depression behavior of a chronic alcohol induction depression model needs more than 6 months.
Drawings
FIG. 1 is a schematic diagram of an alcohol-depression model construction process;
FIG. 2 is a verification result of the tail overhang test;
FIG. 3 is a verification of distance traveled over the entire field of an open field experiment;
FIG. 4 is the time validation results of the peripheral area of the open field experiment;
figure 5 is a verification of forced swim experiments.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Patients with depression often show depressed mood, and alcohol consumption helps to disperse persistent sad mood, while alcohol may temporarily relieve some symptoms of depression, but is likely to cause alcoholism and even develop alcohol dependence; the research also finds that the neurophysiological and metabolic changes caused by the alcoholism are related to the occurrence mechanism of the depression, but the research also shows that the acute alcohol treatment can hardly induce the depression behavior of the mice; while depression may lead a person to a greater risk of developing alcohol dependence, alcohol dependence is more common to develop depression, and alcoholic depressed patients are also now of greater concern.
The existing research finds that the chronic alcohol treatment can induce the depression behavior of the mice for a long time, but the induction period is long, the induction time is usually more than 6 months, and the alcoholic depression model is single in type and is not beneficial to comprehensively understanding the pathogenesis between alcohol and depression, so that the research on the rapid induction alcoholic depression model is beneficial to the research on the potential mechanism of alcohol and depression or the anti-alcoholic depression drugs.
The classical depression model commonly used at present is the LPS acute stress model, which employs LPS-induced treatment to produce depressive behaviour in model animals. LPS induction generally adopts a mode of LPS injection to induce a depression model, such as intraperitoneal injection and intravenous injection, the induction period is usually 4-7 days, although the induction period is short, the mode of LPS injection induction is not consistent with the actual condition of alcoholic depression, the duration of single LPS injection induced depression-like behaviors is short, depression symptoms are light, and alcoholism easily causes severe depression, so that the existing classical LPS induction model is difficult to pertinently simulate the depression-like behaviors of alcoholism people.
However, the experiment shows that the combination of acute alcohol and LPS can stably induce the depression-like behavior of the mouse, the acute alcohol pretreatment of gastric lavage is adopted, and then the combination of LPS induction is adopted, so that the depression behavior of the mouse is induced after 2-4 days, and the depression behavior of the mouse is obvious; furthermore, LPS can be induced by a gastric perfusion method, the method for inducing the depression behavior by the gastric perfusion LPS better meets the actual situation of the depression-like behavior of the alcoholism crowd, and the induction method can better simulate the depression behavior of the alcoholic depression patient.
The invention provides a method for establishing an alcoholic depression animal model, which comprises the following steps:
(1) acute alcohol pretreatment: pretreating multiple times of gavage experimental animals by using acute alcohol with a preset gavage amount, wherein the preset gavage amount is calculated by 5-10g/kg of each gavage according to the weight of the experimental animals; the multiple times of gavage, preferably 3-5 times of gavage, are performed at intervals of 12-18h each time, and are used for simulating the drinking behavior of the alcoholism crowd;
(2) LPS induced depression model: inducing the experimental animal by adopting LPS 2-6h after the acute alcohol pretreatment in the step (1) until a depression-like behavior appears, and verifying the constructed alcoholic depression model; the constructed alcoholic depression model can be subjected to model verification through behavior detection, such as open field experiments, tail suspension experiments and forced swimming experiments.
Preferably, the induction is carried out by means of single-time gavage LPS, and the weight of each gavage is 10-15mg/kg according to the weight of experimental animals.
Acute alcohol gavage pretreatment is combined with LPS induction, depression-like behaviors of the mice can be induced within 2-3 days, and compared with the induction period of chronic alcohol, the method can greatly shorten the experimental modeling time; in addition, LPS in a classical depression modeling mode is induced by a mode of intravenous injection or intraperitoneal injection, a depression model can be induced by a mode of intragastric administration LPS after acute alcohol is firstly used for multiple times of intragastric administration, the mode of intragastric administration LPS induction better accords with the actual situation of alcoholic depression, depression-like behaviors of people caused by alcoholism can be better simulated, the potential mechanism of the relation between alcohol and depression can be favorably researched, the depression behaviors of mice are obvious, and the model effect is good.
In addition, the invention also provides a depression model constructed by the alcoholic depression model establishing method.
In addition, the invention also provides application of the model obtained by the alcoholic depression model establishing method in screening anti-alcoholic depression drugs.
The following are examples:
example 1 Induction of alcohol Depression model by alcohol + LPS (mLPS model)
The flow chart of the method for constructing the alcoholic depression model is shown in fig. 1, and the specific steps are as follows:
(1) alcohol pretreatment: carrying out acute alcohol intragastric administration for three times on each mouse, wherein the intragastric administration amount of each mouse is 5g/kg according to the weight of the mouse, and the intragastric administration interval is 12 h; saline was used as a blank control.
(2) The lavage LPS induced depression model: 2h after the third intragastric administration of the acute alcohol, performing LPS single intragastric administration on the mice, wherein the intragastric administration amount of each intragastric administration is 10mg/kg according to the weight of the mice; the behavioral tests were performed on the mice on days 3-4, and the results are shown in the mALPS groups in FIGS. 2-5.
COMPARATIVE EXAMPLE 1 alcohol-induced Depression model (EtOH model)
The experimental method comprises the following steps: carrying out three times of acute alcohol gavage (5g/kg/dose) on each mouse, and carrying out gavage on a control group with the same amount of physiological saline, wherein the interval of each gavage is 12 hours; after the acute alcohol intragastric administration for the third time is carried out for 2 hours, the physiological saline with the same amount of intragastric administration is taken as the physiological saline with the intragastric administration amount of 10 mg/kg; the mouse behaviours were examined on days 3-4 and the results are shown in the EtOH groups in FIGS. 2-5.
Comparative example 2 model of Depression induced by intragastric LPS
The experimental method comprises the following steps: and carrying out three times of normal saline intragastric administration on each mouse, wherein the intragastric administration amount is 5g/kg/dose, carrying out LPS single intragastric administration 2 hours after the third time of normal saline intragastric administration, and carrying out behavioral detection on the mice on 3-4 days by using the normal saline with the same amount as the intragastric administration of each control group according to the weight of the mice, wherein the results are shown in LPS groups in the figures 2-5.
Fig. 2-5, wherein fig. 2 is the verification result of the tail suspension test, fig. 3 is the verification result of the distance traveled by the whole area of the open field test, fig. 4 is the verification result of the time of the peripheral area of the open field test, and fig. 5 is the verification result of the forced swimming test.
As shown in fig. 2-5, compared to the control group, the mice showed no depression-like behavior in both the gavage acute alcohol and the gavage LPS group alone, while the mice in the mahps group (acute alcohol + LPS) showed significant depression-like behavior, indicating that the acute alcohol + LPS induced depression model in mice was successfully constructed, while the gavage acute alcohol or the gavage LPS induced depression model failed.
In conclusion, the mine field experiment, the tail suspension experiment and the forced swimming experiment are verified to show that the alcoholic depression model can be constructed by the method, the behavior of the mouse depression sample is obvious, and the model effect is good.
It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (8)
1. A method for establishing an alcoholic depression animal model is characterized by comprising the following steps:
selecting experimental animals meeting experimental requirements;
pretreating an experimental animal subjected to multiple times of intragastric administration with acute alcohol with a preset intragastric administration amount; the preset gavage amount is calculated by 5-10g/kg of gavage of each experimental animal according to the weight of the experimental animal;
and (3) after the acute alcohol pretreatment, inducing the experimental animal by using LPS until a depression-like behavior appears, and verifying the constructed alcoholic depression model.
2. The method for establishing animal model of alcoholic depression according to claim 1, wherein said LPS induction is induced by means of intragastric LPS, and is measured in terms of per-animal intragastric administration at 10-15mg/kg according to the body weight of the experimental animal.
3. The method for establishing an animal model of alcoholic depression according to claim 1, wherein the multiple gavages are gavages 3-5 times with an interval of 12-18 h.
4. The method for establishing an animal model of alcoholic depression according to claim 3, wherein LPS is further used to induce the experimental animal 2-6 hours after the acute alcohol pretreatment.
5. The method for establishing animal model of alcoholic depression according to claim 4, wherein the acute alcohol is gavaged 3 times at intervals of 12h, in terms of the experimental animal body weight, in 5 g/kg/time per gavage.
6. The method for establishing animal model of alcoholic depression according to claim 5, wherein said LPS induction is a single induction, and is performed in an amount of 10mg/kg per gavage according to the body weight of the experimental animal.
7. A depression model constructed by the method for constructing alcoholic depression model according to any one of claims 1 to 6.
8. The use of the depression model constructed by the alcoholic depression model establishing method as claimed in any one of claims 1 to 6 in screening anti-alcoholic depression drugs.
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