阅读说明：本技术 大鼠恐惧应激与气态污染物复合暴露模型的建立方法 (Method for establishing rat fear stress and gaseous pollutant composite exposure model ) 是由 田蕾 刘晓华 袭著革 李康 杲修杰 刘焕亮 林本成 来文庆 于 2020-04-24 设计创作，主要内容包括：本发明属于动物模型建立技术领域,本发明公开了一种大鼠恐惧应激与气态污染物复合暴露模型的建立方法。所述的方法包括如下步骤：(1)建立恐惧模型：选取180-220g健康雄性wistar大鼠,置于密闭环境中,给予75dB:1000Hz声音信号,在声音信号末尾3s同时给予1mA足底电流刺激,每日重复周期20次,其余时间正常饲养自由食水；持续六周；(2)建立恐惧应激与气态污染物复合暴露模型：将完成恐惧模型建立的大鼠进行气态污染物吸入,每天吸入时间为4小时,持续30天；(3)采样检测。本发明的方法复合了两种有害因素,对人体毒性危害进行评价。(The invention belongs to the technical field of animal model establishment, and discloses a method for establishing a rat fear stress and gaseous pollutant composite exposure model. The method comprises the following steps: (1) establishing a fear model: selecting 180-220g healthy male wistar rats, placing the rats in a closed environment, giving a 75dB 1000Hz sound signal, giving 1mA plantar current stimulation at the same time for 3s at the tail of the sound signal, repeating the cycle for 20 times every day, and normally feeding free edible water in the rest time; lasting for six weeks; (2) establishing a fear stress and gaseous pollutant composite exposure model: sucking gaseous pollutants into the rat which is built by the fear model, wherein the sucking time is 4 hours every day and lasts for 30 days; (3) and (5) sampling and detecting. The method of the invention combines two harmful factors and evaluates the toxicity hazard of human body.)

1. A method for establishing a rat fear stress and gaseous pollutant compound exposure model is characterized by comprising the following steps:

(1) establishing a fear model: selecting 180-220g healthy male wistar rats, placing the rats in a closed environment, giving a 75dB 1000Hz sound signal, giving 1mA plantar current stimulation at the same time for 3s at the tail of the sound signal, repeating the cycle for 20 times every day, and normally feeding free edible water in the rest time; lasting for six weeks;

(2) establishing a fear stress and gaseous pollutant composite exposure model: sucking gaseous pollutants into the rat which is built by the fear model, wherein the sucking time is 4 hours every day and lasts for 30 days;

(3) and (5) sampling and detecting.

2. The method of claim 1 wherein said gaseous contaminant is xylene.

3. The method of claim 2, wherein the concentration of xylene inhaled by rats is 0.38 ppm.

4. The method of claim 2, wherein the concentration of xylene inhaled by rats is 3.8 ppm.

5. The method of claim 1, wherein said sample testing comprises general behavior, percent freezing time, central nervous system, immune system, and endocrine system.

Technical Field

The invention relates to the technical field of animal model establishment, in particular to a method for establishing a rat fear stress and gaseous pollutant composite exposure model.

Background

The description of the background of the invention pertaining to the related art to which this invention pertains is given for the purpose of illustration and understanding only of the summary of the invention and is not to be construed as an admission that the applicant is explicitly or implicitly admitted to be prior art to the date of filing this application as first filed with this invention.

Various physical and chemical harmful factors in the closed space of the submarine cabin are various and exist at the same time. The influence of different degrees on all aspects can be caused to the sailor, the change of physiological indexes and the appearance of psychological problems can be caused, and various diseases can be caused.

Harmful gas in submarine operation environment is generated from nonmetal materials used by submarines in large quantity, cooking food, self metabolites of the submariners, machine operation and the like, such as carbon monoxide, dimethylbenzene, acrolein, hydrogen sulfide and the like.

The submarine crew is in danger of operation in a closed space in deep sea, various training and sudden conditions inside and outside the submarine when performing tasks, particularly long-time deep diving tasks, and psychological loads of fear stress states exist, and physical and psychological influences are generated by the coexistence of other harmful factors.

Disclosure of Invention

The invention aims to provide a method for establishing a rat fear stress and gaseous pollutant compound exposure model.

A method for establishing a rat fear stress and gaseous pollutant compound exposure model, which comprises the following steps:

(1) establishing a fear model: selecting 180-220g healthy male wistar rats, placing the rats in a closed environment, giving a 75dB 1000Hz sound signal, giving 1mA plantar current stimulation at the same time for 3s at the tail of the sound signal, repeating the cycle for 20 times every day, and normally feeding free edible water in the rest time; lasting for six weeks;

(2) establishing a fear stress and gaseous pollutant composite exposure model: sucking gaseous pollutants into the rat which is built by the fear model, wherein the sucking time is 4 hours every day and lasts for 30 days;

(3) and (5) sampling and detecting.

Further, the gaseous contaminant is xylene.

Further, the concentration of xylene inhaled by rats was 0.38 ppm.

Further, the concentration of xylene inhaled by rats was 3.8 ppm.

Further, the sampling test includes general behaviors, percent freezing time, central nervous system, immune system and endocrine system.

The invention has the following beneficial effects:

the method builds a stable and long-acting conditioned reaction fear model, realizes the compounding of two harmful factors by correlating the contamination and fear through specific conditioned signals, compounds the fear stress and the main gaseous pollutants in the submarine closed space working environment, researches the influence of harmful gas inhalation on the health conditions of animal physiological behaviors and the like under the fear stress state, and further discusses the evaluation of the toxicity hazard to human bodies.

Drawings

FIG. 1 is a graph of percent freezing time (FT%) for control group C versus model group F in accordance with the present invention;

FIG. 2 is a graph showing the weight gain of the control group C and the model group F according to the present invention;

FIG. 3 is a graph showing a comparison between the trajectory display and exploration behaviors of a control group C and a model group F according to the present invention;

FIG. 4 is a graph showing the results of the test of the effect of fear stress on the body according to the present invention;

FIG. 5 is a graph showing the results of the test of the effect of fear stress on the body according to the present invention;

FIG. 6 is a graph showing the results of the test of the effect of fear stress on the body according to the present invention;

FIG. 7 is a graph showing the results of the test of the effect of fear stress on the body according to the present invention;

FIG. 8 is a graph showing the results of measurement of the influence of fear stress on arachidonic acid metabolism according to the present invention;

FIG. 9 is a graph showing the results of pathological examination of hippocampus in accordance with the present invention;

FIG. 10 is a diagram showing the results of pathological examination of cerebral cortex according to the present invention;

FIG. 11 is a diagram showing the results of the amygdala assay in the brain of the present invention;

FIG. 12 is a graph showing the results of pharmacological tests of the present invention;

FIG. 13 is a graph showing the results of pathological liver tests according to the present invention;

FIG. 14 is a graph showing the results of measurement of Corticotropin Releasing Hormone (CRH) in blood according to the present invention;

FIG. 15 is a graph showing the results of detection of Corticosterone (CORT) in blood according to the present invention;

FIG. 16 is a graph showing the results of measurement of adrenocorticotropic hormone (ACTH) in blood according to the present invention;

FIG. 17 is a graph showing the results of the Norepinephrine (NE) detection in blood according to the present invention;

FIG. 18 is a graph showing the result of detecting Acetylcholine (ACH) in brain tissue according to the present invention;

FIG. 19 is a graph showing the result of Dopamine (DA) detection in brain tissue according to the present invention;

FIG. 20 is a graph showing the results of 5-hydroxytryptamine (5-HT) assay in brain tissue according to the present invention.

Detailed Description

The present application is further described below with reference to examples.

In the following description, different "one embodiment" or "an embodiment" may not necessarily refer to the same embodiment, in order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art. Various embodiments may be replaced or combined, and other embodiments may be obtained according to the embodiments without creative efforts for those skilled in the art.

A method for establishing a rat fear stress and gaseous pollutant compound exposure model, which comprises the following steps:

(1) establishing a fear model: selecting 180-220g healthy male wistar rats, placing the rats in a closed environment, giving a 75dB 1000Hz sound signal, giving 1mA plantar current stimulation at the same time for 3s at the tail of the sound signal, repeating the cycle for 20 times every day, and normally feeding free edible water in the rest time; lasting for six weeks;

(2) establishing a fear stress and gaseous pollutant composite exposure model: sucking gaseous pollutants into the rat which is built by the fear model, wherein the sucking time is 4 hours every day and lasts for 30 days;

(3) and (5) sampling and detecting.

The first stage uses fear experimental box to make the mould, is in relative confined environment with the experimental animal, gives the sound signal of specific frequency, and the vola electro photoluminescence is given simultaneously to audio frequency end several seconds, makes the animal produce the fear state. The animal produces the fear of condition reaction in the molding process, and this kind of specific audio frequency is the condition that the induced animal appears the fear state, hears this audio frequency promptly and will produce the fear state. Through long-time modeling experiments, the conditioned reflex is reinforced, and relatively long-time memory is formed, so that the time of the next compound infection experiment is facilitated.

In some embodiments of the invention, the gaseous contaminant is xylene.

In some embodiments of the invention, the concentration of xylene inhaled by rats is 0.38 ppm.

In some embodiments of the invention, the concentration of xylene inhaled by rats is 3.8 ppm.

In some embodiments of the invention, the sampling test includes general behavior, percent freezing time, central nervous system, immune system, and endocrine system.