Experimental device and method for simulating rat reproduction influence under exposure of low-pressure anoxic environment

Abstract

The invention discloses an experimental device and method for simulating rat reproduction influence under exposure of a low-pressure anoxic environment, relates to the technical field of reproduction influence experiments, and provides the following scheme that the experimental device comprises a low-pressure experimental module fixed on the ground and a vacuum pump erected on the top surface of the low-pressure experimental module, the extraction end of the vacuum pump penetrates into an inner cavity of the low-pressure experiment module through a pipeline, a hatch is formed in the left side of the low-pressure experiment module, and a hatch door is installed on the end face of the hatch through a hinge. According to the technical scheme, by simulating the day and night temperature difference at the altitude of 4000 m and 6000 m and 12 DEG C / 2 DEG C in the low-pressure experiment module and simulating the plateau environment to feed rats, the plateau low-oxygen exposure environment can be closer to the plateau low-oxygen exposure environment, and by conducting slice detection on testicular tissue of the test rats, damage data of the low-pressure low-oxygen exposure environment to the male reproductive system is obtained. Therefore, the method has certain good reproducibility on reproductive system injury in a low-pressure and low-oxygen exposure environment.

Description

Technical Field

[0001] The invention relates to the technical field of reproductive impact experiments, and in particular to an experimental device and method for simulating reproductive impact on rats under exposure to a low-pressure hypoxic environment. Background Art

[0002] Some people who have lived in plains for a long time and cannot adapt to the hypoxic environment may experience a series of non-specific symptoms when they enter areas above 3000m in a short period of time. In some severe cases, they may even rapidly develop into acute mountain sickness. AMS refers to various clinical syndromes that occur in a short period of time (several hours to several days) when people enter the plateau from the plain or from the plateau to higher altitude areas. It is the main disease that affects the physical health and life safety of people who quickly enter high-altitude areas. In recent years, there have been limited studies on the effects of plateau hypoxia on the male reproductive system. At present, there are few experimental studies that use volunteers as subjects to enter a hypoxic environment to simulate AMS. Previous studies have shown that the plateau hypoxic environment can cause damage to the male testicular reproductive function, but the specific mechanism is not yet fully clear. Constructing a plateau hypobaric hypoxia reproductive system damage model is one of the key links in the study of plateau-related reproductive system diseases. Summary of the invention

[0003] In view of the defect that the above background technology raises whether the high-altitude hypoxic environment can cause damage to the male testicular reproductive function, and the specific mechanism is not yet fully clear, a technical solution of an experimental device for simulating the reproductive effects of rats under exposure to a low-pressure hypoxic environment is provided; it includes a low-pressure experimental cabin fixed on the ground, and a vacuum pump mounted on the top surface of the low-pressure experimental cabin, wherein the extraction end of the vacuum pump penetrates into the inner cavity of the low-pressure experimental cabin through a pipeline, and a hatch is opened on the left side of the low-pressure experimental cabin, and a hatch door is installed on the end face of the hatch through a hinge.

[0004] Preferably, two lighting observation windows are arranged on the rear side wall of the low-pressure experimental chamber, and several groups of male experimental rats are raised inside the low-pressure experimental chamber.

[0005] An experimental method for simulating the reproductive effects of rats exposed to a low-pressure hypoxic environment includes the following experimental steps:

[0006] S1: The rats were placed in a hypobaric cabin with the door sealed. A vacuum pump was used to partially extract the air in the cabin to simulate the low-pressure and low-oxygen environment of the plateau. The pressure was reduced at a speed of 2 m / s. The plateau model group I simulated an ascent to an altitude of 4000 m, and the plateau model group II simulated an ascent to an altitude of 6000 m. Fresh air was circulated every 4-6 hours. The oxygen cabin maintained a temperature of 22-24°C. The rats were raised for 14 days, during which time they had free access to food and water. After the experimental time was completed according to the experimental groups, the air intake valve was adjusted. After the experiment, the rats descended to the normal altitude at a speed of 2 m / s.

[0007] S2: After the feeding experiment, the rats in each group were taken out of the cabin and immediately anesthetized with an intraperitoneal injection of 10% chloral hydrate at a dose of 1 ml / kg. The rats were placed in a supine position, and the abdominal aorta was isolated after the abdominal cavity was exposed. About 1 ml of abdominal aortic blood was drawn and placed in a heparin anticoagulation tube, and blood gas analysis was performed within 5 minutes.

[0008] S3: Testicular tissue sampling: The testicular tissue of rats was peeled off to observe the gross pathological changes of testicular tissue. The testicular tissue of the left rat was fixed with 10% formaldehyde for 24 h, embedded in paraffin, sliced ​​at 4-6 μm, mounted on clean slides, routinely dewaxed and dehydrated, dehydrated and mounted after HE staining, and observed under a light microscope;

[0009] S4: The epididymal tails of rats in each group were washed with PBS, cut into pieces with ophthalmic scissors, and shaken evenly. After standing for 20 minutes, they were filtered. After standing for 3 minutes, the supernatant was taken to prepare sperm suspension for the detection of sperm motility, concentration, and deformity rate.

[0010] S5: Take out the left testicle on an ice box, rinse and weigh it, cut the tissue into pieces and make a homogenate, centrifuge the prepared 10% homogenate, and save the supernatant for later use.

[0011] S6: SPSS20.0 statistical software was used for data statistics. The quantitative data that conformed to the normal distribution and had equal variance were expressed as x±s. Factorial design variance analysis was used to compare the blood gas, sperm motility, concentration, deformity rate, MDA, and SOD of rats at different simulated altitudes.

[0012] Preferably, the rats are kept in an environment with a temperature of 21-24°C, a humidity of 50%-60%, a 12-hour day and night rhythm, free access to food and water, and are randomly divided into 3 groups, 20 in each group, and kept for 14 days; the plain group (local altitude) is kept in a clean room of an animal center outside the cabin and subjected to normoxic treatment, and the model group is divided into two groups, plateau model group I (4000m above sea level) and plateau model group II (6000m above sea level).

[0013] Preferably, during the ventilation process in step S1, the pressure in the cabin needs to be kept stable.

[0014] Preferably, in step S6, one-way analysis of variance is used to compare the means of multiple samples, and a t-test is used to compare the means of two samples.

[0015] It can be seen from the above technical scheme that the present invention provides an experimental device and method for simulating the reproductive effects of rats exposed to a low-pressure hypoxic environment. Compared with the prior art, the present invention has the following beneficial effects:

[0016] The technical solution of the present invention simulates an altitude of 4000 or 6000 m and a day and night temperature difference of 12°C / 2°C in a low-pressure experimental chamber, and raises rats in a simulated plateau environment, which can be closer to the plateau hypoxic exposure environment. By performing section tests on the testicular tissue of the experimental rats, data on the damage to the male reproductive system in the low-pressure and low-oxygen exposure environment are obtained, thereby having a certain good reproducibility of the damage to the reproductive system in the low-pressure and low-oxygen exposure environment. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces and describes the drawings required for use in the embodiments of the present invention or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention, and for ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0018] Figure 1 This is the overall schematic diagram of the low-pressure hypoxic environment experimental device;

[0019] Figure 2 This is a schematic diagram of the low-pressure hypoxic environment experimental device after cutting;

[0020] Figure 3 Schematic diagram of testicular histopathological changes in rats in each group;

[0021] Figure 4 Schematic diagram of arterial blood gas analysis of rats in each group;

[0022] Figure 5 Schematic diagram of the comparison of sperm parameters among the groups;

[0023] Figure 6 Schematic diagram of the comparison of MDA and SOD levels in the testicular tissue of rats in each group.

[0024] Attached Figure 1 -Attached Figure 2 The corresponding relationship of the parts is as follows:

[0025] 1. Low-pressure experimental chamber; 2. Vacuum pump; 3. Hatch door; 4. Hatch opening; 5. Lighting observation window. DETAILED DESCRIPTION

[0026] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described below are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] In order to more clearly explain and illustrate the technical solution and implementation of the present invention, several preferred specific embodiments for implementing the technical solution of the present invention are introduced below.

[0028] Example

[0029] A preferred technical solution for an experimental device simulating the reproductive effects of rats exposed to a low-pressure hypoxic environment: comprising a low-pressure experimental chamber 1 fixed on the ground, and a vacuum pump 2 mounted on the top surface of the low-pressure experimental chamber 1, wherein the extraction end of the vacuum pump 2 penetrates into the inner cavity of the low-pressure experimental chamber 1 through a pipeline, and a hatch 4 is opened on the left side of the low-pressure experimental chamber 1, and a hatch door 3 is installed on the end surface of the hatch 4 through a hinge. Two lighting observation windows 5 are arranged on the rear side wall of the low-pressure experimental chamber 1, and several groups of male experimental rats are raised inside the low-pressure experimental chamber 1. A sterilizable air pressure buffer cabin is designed for the cabin door, in which the animals can be left first, and the pressure is slowly increased to 3000m after 15 minutes, and then the cabin pressure is simultaneously increased to 3000m, and the buffer cabin door is opened to drive the animals into the cabin, and then the door is closed and the pressure is gradually increased to the air pressure required for the experiment. A complete set of automatic animal feeding settings (drinking water, eating, and feces cleaning) are designed in the cabin, and the operation and settings are completed by the color LCD touch screen outside the cabin.

[0030] A technical solution for an experimental method simulating the reproductive effects of rats exposed to a low-pressure hypoxic environment:

[0031] Experimental Animals and Groups Sixty SPF Wistar male rats, 24 weeks old,

[0032] Rats weighing (200±20)g were kept in a clean room in the animal center, with a temperature of 21-24℃, a humidity of 50%-60%, a 12-hour day and night rhythm, and free access to food and water. They were randomly divided into 3 groups, 20 in each group, and raised for 14 days. The plain group (local altitude) was raised in a clean room outside the cabin animal center and treated with normoxia. The model group was divided into two groups, plateau model group I (4000m above sea level) and plateau model group II (6000m above sea level).

[0033] Experimental equipment and reagents: simulated high-altitude low-pressure and low-oxygen animal experimental chamber, desktop high-speed centrifuge, animal complete blood cell analyzer, MDA, SOD kit.

[0034] The rat model of reproductive system injury in the hypobaric chamber was established; the animal experiment chamber simulating plateau hypobaric hypoxia was used to simulate different altitudes for animal experiments. The rats were placed in the hypobaric chamber, the cabin door was sealed, and the air in the cabin was partially extracted by a vacuum pump to simulate the low-pressure and low-oxygen environment of the plateau. The pressure was reduced at a speed of 2m / s. The plateau model group I simulated an ascent to an altitude of 4000m, and the plateau model group II simulated an ascent to an altitude of 6000m. Fresh air was circulated every 4 to 6 hours (the pressure in the cabin was kept stable during the ventilation process), and the oxygen chamber maintained a temperature of 22 to 24℃. The rats were fed for 14 days, during which time they were free to eat and drink. After the experimental time was completed according to the experimental groups, the air intake valve was adjusted. After the experiment, they descended to the normal altitude at a speed of 2m / s.

[0035] For blood gas determination, after the feeding experiment, the rats in each group were taken out of the cabin and immediately anesthetized with an intraperitoneal injection of 10% chloral hydrate at a dose of 1 ml / kg. The rats were placed in a supine position, and the abdominal aorta was isolated after the abdominal cavity was exposed. About 1 ml of abdominal aortic blood was drawn and placed in a heparin anticoagulant tube, and blood gas analysis was performed within 5 minutes.

[0036] Testicular tissue sampling, rat testicular tissue peeling, and observation of gross pathological changes in testicular tissue. The left rat testicular tissue was fixed with 10% formaldehyde for 24 hours, embedded in paraffin, sliced ​​at 4-6 μm, mounted on clean slides, routinely dewaxed and dehydrated, dehydrated and sealed after HE staining, and observed under a light microscope.

[0037] For the detection of semen parameters, the cauda epididymis on both sides of the rats in each group were taken, washed with PBS, cut into pieces with ophthalmic scissors and shaken evenly, allowed to stand for 20 minutes and then filtered. After filtration, allowed to stand for 3 minutes, the supernatant was taken to prepare sperm suspension for the detection of sperm motility, concentration, and deformity rate.

[0038] MDA and SOD levels were tested on an ice box according to the instructions. The left testicle was removed, rinsed, weighed, and the chopped tissue was made into a homogenate. The prepared 10% homogenate was centrifuged and the supernatant was saved for later use.

[0039] SPSS20.0 statistical software was used for data statistics. Measurement data that conformed to normal distribution and homogeneous variance were expressed as x±s. Factorial design variance analysis was used to compare blood gas, sperm motility, concentration, deformity rate, MDA, and SOD in rats at simulated different altitudes. One-way variance analysis was used to compare the means of multiple samples, and t-test was used to compare the means of two samples. P≤0.05 was considered statistically significant.

[0040] Arterial blood gas changes Blood gas analysis results showed that compared with the plain group, the plateau model group had a significant decrease in ⅠPO2 (P < 0.05), HCT, Cl － , Hb significantly increased (P<0.01); in plateau model group Ⅱ, pH and PO2 significantly decreased (P<0.01), PCO2, HCT, K+ , Cl － , Hb were significantly increased (P<0.01); Na+ was increased in both plateau model group Ⅰ and plateau model group Ⅱ, but the difference was not statistically significant (P>0.05).

[0041] Pathological changes of testicular tissue were observed under light microscope. The basement membrane of seminiferous tubules in the testicular tissue of rats in the plain group was intact, and the whole process of spermatogenesis was visible. There were more sperm in the tubular cavity, and the sperm was not obviously damaged. The cells were arranged neatly, about 5-7 layers, and the cell polarity was regular. Compared with the plain group, the basement membrane of seminiferous tubules in the testicular tissue of the plateau model group I had slight changes, but not obvious, showing mild wrinkles, atrophy, and stratification. The number of sperm increased slightly, the spermatogenesis was not obviously damaged, mature spermatogenic cells were lacking, and the cell arrangement was slightly Compared with the plain group, the basement membrane of the seminiferous tubules of the testicular tissue of the plateau model group Ⅱ rats was atrophied and thinned, with obvious wrinkles and stratification, obvious vacuoles, very few sperm, a few early spermatogenic cells, extremely disordered cell arrangement, reduced layers, and cell polarity disappeared. 2.3 Changes in semen parameters Compared with the plain group, the sperm motility rate of the plateau model group Ⅰ was significantly reduced (P < 0.01), and the sperm deformity rate was significantly increased (P < 0.01). Compared with the plain group, the sperm motility rate of the plateau model group Ⅱ was significantly reduced (P < 0.01), the sperm deformity rate was significantly increased (P < 0.01), and the sperm concentration was significantly decreased (P < 0.01).

[0042] Compared with the plain group, the MDA concentration of the testicular tissue homogenate of the rats in the plateau model group I was significantly increased (P < 0.01), and the SOD activity was significantly decreased (P < 0.05). Compared with the plain group, the MDA concentration of the testicular tissue homogenate of the rats in the plateau model group II was significantly increased (P < 0.01), and the SOD activity was significantly decreased (P < 0.01).

[0043] Plateau refers to areas with an altitude of more than 3000m. Its typical climatic characteristics include high cold, low oxygen partial pressure and high solar radiation. In recent years, a large number of studies have shown that the hypoxic environment of the plateau has considerable damage to various organs and systems of the body. Hypoxic exposure can trigger a series of complex chain reactions in the body at the endocrine, cellular and genetic levels. The first problem to be solved in the study of plateau reproductive injury is the replication of the animal model of acute plateau reproductive system injury.

[0044] At present, the diagnosis of AMS in the human population adopts the Lake Louise scoring standard or the domestic AMS disease diagnostic standard, which is mainly based on a series of symptoms that appear in the plateau hypoxic environment, and is diagnosed by quantitative scoring. However, after the animal model is replicated, because it is impossible to score and quantify the symptoms of animals like humans, other indicators need to be selected to judge the success of the animal acute mountain sickness model. At the same time, the selection of animal age is also very important. It is necessary to select experimental animals of similar age to adults, whose male reproductive system is mature and suitable for simulating reproductive system damage under low pressure and hypoxia in the plateau. Therefore, the Wistar male rats selected in this study are 24 weeks old, with a well-developed reproductive system and can complete autonomous mating, which is 18 to 20 years old compared to the human age.

[0045] Blood gas analysis is the main means of evaluating the body's hypoxia and acid-base imbalance. In medicine, it is often used as a test method to determine whether the body has acid-base imbalance and hypoxia and the degree of hypoxia. In this experiment, some indicators have changed when simulating an altitude of 4000m, PO2 decreased, HCT, Cl － , Hb increased; while when simulating entering the plateau at an altitude of 6000m, pH and PO2 decreased, PCO2, HCT, K + , Cl － , Hb increased. The higher the simulated altitude, the more obvious the decrease in oxygen partial pressure and the increase in CO2. This can be used to evaluate the hypoxia condition of rats in the early stage of hypoxia.

[0046] Testicular tissue pathological examination is the gold standard for diagnosing testicular spermatogenesis. This study found that at a simulated altitude of 4000m, the basement membrane of the seminiferous tubules of the testicular tissue had slight changes, but not obvious, showing mild wrinkles, atrophy, and stratification, a small increase in the number of sperm, and no obvious damage to spermatogenesis. There was a lack of mature spermatogenic cells, and the cell arrangement was slightly disordered and sparse, with 3-5 layers, and cell polarity was visible; while at a simulated altitude of 6000m, the basement membrane of the seminiferous tubules atrophied and thinned, with obvious wrinkles and stratification, obvious vacuoles, very few sperm, a few early spermatogenic cells, extremely disordered cell arrangement, reduced layers, and cell polarity disappeared. These changes will destroy the normal structure and function of germ cells and lead to damage to reproductive function. The above results show that the simulated plateau environment has little effect on the morphology of the testicular seminiferous tubules of rats at an altitude of 4000m, but it is most obvious at an altitude of 6000m.

[0047] Some studies have shown that the low-pressure and low-oxygen environment at high altitudes has a significant impact on the male reproductive system, which can lead to a decrease in the number of male sperm, a decrease in sperm motility and viability, an increase in sperm deformity, an increase in germ cell apoptosis, and a disorder in reproductive hormone secretion. The experiment found that with the increase in altitude, sperm motility and concentration decreased significantly, and sperm deformity increased significantly, which was most obvious at an altitude of 6000m, indicating that the low-pressure and low-oxygen environment does have a significant impact on the spermatogenesis of rats, and the higher the altitude, the more obvious the damage.

[0048] ROS mainly participates in the occurrence and development of many chronic progressive diseases by damaging DNA bases, protein oxidation products, lipid peroxidation products, etc. Under normal physiological conditions, oxygen free radicals in the body are in a dynamic equilibrium state, with oxygen free radicals being continuously generated and removed.

[0049] However, under the influence of certain external factors, the body may produce more oxygen free radicals or reduce the speed of removing oxygen free radicals, which will lead to oxidative stress. Through experiments, we found that the increase of MDA in tissues and the decrease of antioxidant SOD indicate that the production and removal of oxygen free radicals in rat testicular tissues have been unbalanced in the simulated high-altitude environment. Some foreign studies have also confirmed this point. Intermittent hypoxia leads to an increase in serum lipid peroxide MDA and a decrease in total antioxidant capacity, a decrease in the activity of antioxidants SOD, GSH-Px, and GSR in the testis and epididymis, and a significant decrease in epididymal sperm count. It was also found that this situation can be improved by not supplementing ascorbic acid and polyunsaturated fatty acids.

[0050] In summary, the exploration of the evaluation indicators for the replication of the acute high altitude rat reproductive system injury model showed that oxygen partial pressure, testicular histopathological morphology, sperm motility, sperm deformity rate, MDA, and SOD had changed when the altitude was simulated at 4000m, and it was more obvious when the altitude was simulated at 6000m. At the same time, at an altitude of 6000m, pH, PO2, PCO2, HCT, and K + , Cl － , Hb, and sperm counts all changed, indicating that being in a plateau environment at an altitude of 6000m may have a more serious impact on the reproductive system function of rats. By studying the effects of hypoxia at an altitude of 5000m for 5, 15, and 30 days on the reproductive system of rats, it was found that the apoptosis rate of germ cells was the highest at 15 days, and the number of sperm in the epididymis was also the lowest at 15 days. The method of making the model in this study: 4000, 6000m, 12℃ / 2℃ day and night temperature difference, simulating the plateau environment for 14 days, can be closer to the plateau hypoxic exposure environment, which may be more beneficial to reflect the damage of low-pressure and low-oxygen exposure to the male reproductive system, and it has been tested that this model has a certain good reproducibility.

[0051] The present invention is not limited to the above-mentioned optimal implementation mode. Anyone should be aware that any structural changes made under the inspiration of the present invention, and any technical solutions that are the same or similar to the present invention, fall within the protection scope of the present invention. Finally, it should be noted that the structures, proportions, sizes, etc. illustrated in the drawings of this specification are only used to match the contents disclosed in the specification for people familiar with this technology to understand and read, and are not used to limit the restrictive conditions that can be implemented in this application, so they have no technical substantive significance. Any structural modification, change in proportional relationship or adjustment of size, without affecting the effects that can be produced by this application and the purposes that can be achieved, should still fall within the scope that can be covered by the technical content disclosed in this application.

Claims

1. An experimental device for simulating the reproductive effects of rats exposed to a low-pressure hypoxic environment, characterized in that: The invention comprises a low-pressure test chamber (1) fixed on the ground, and a vacuum pump (2) mounted on the top surface of the low-pressure test chamber (1), wherein the extraction end of the vacuum pump (2) penetrates into the inner cavity of the low-pressure test chamber (1) through a pipeline, and a hatch (4) is opened on the left side of the low-pressure test chamber (1), and a hatch door (3) is installed on the end surface of the hatch (4) through a hinge.

2. The experimental device for simulating the reproductive effects of rats under low-pressure hypoxia environment exposure according to claim 1, characterized in that: Two lighting observation windows (5) are arranged on the rear side wall of the low-pressure experimental chamber (1), and several groups of male experimental rats are raised inside the low-pressure experimental chamber (1).

3. An experimental method for simulating the reproductive effects of rats exposed to a low-pressure hypoxic environment, characterized in that: An experimental device for simulating the reproductive effects of rats exposed to a low-pressure hypoxic environment as described in any one of claims 1-2, and comprising the following experimental steps: S1: The rats were placed in a hypobaric cabin with the door sealed. A vacuum pump was used to partially extract the air in the cabin to simulate the low-pressure and low-oxygen environment of the plateau. The pressure was reduced at a speed of 2 m / s. The plateau model group I simulated an ascent to an altitude of 4000 m, and the plateau model group II simulated an ascent to an altitude of 6000 m. Fresh air was circulated every 4-6 hours. The oxygen cabin maintained a temperature of 22-24°C. The rats were raised for 14 days, during which time they had free access to food and water. After the experimental time was completed according to the experimental groups, the air intake valve was adjusted. After the experiment, the rats descended to the normal altitude at a speed of 2 m / s. S2: After the feeding experiment, the rats in each group were taken out of the cabin and immediately anesthetized with an intraperitoneal injection of 10% chloral hydrate at a dose of 1 ml / kg. The rats were placed in a supine position, and the abdominal aorta was isolated after the abdominal cavity was exposed. About 1 ml of abdominal aortic blood was drawn and placed in a heparin anticoagulation tube, and blood gas analysis was performed within 5 minutes. S3: Testicular tissue sampling: The testicular tissue of rats was peeled off to observe the gross pathological changes of testicular tissue. The testicular tissue of the left rat was fixed with 10% formaldehyde for 24 h, embedded in paraffin, sliced ​​at 4-6 μm, mounted on clean slides, routinely dewaxed and dehydrated, dehydrated and mounted after HE staining, and observed under a light microscope; S4: The epididymal tails of rats in each group were washed with PBS, cut into pieces with ophthalmic scissors, and shaken evenly. After standing for 20 minutes, they were filtered. After standing for 3 minutes, the supernatant was taken to prepare sperm suspension for the detection of sperm motility, concentration, and deformity rate. S5: Take out the left testicle on an ice box, rinse and weigh it, cut the tissue into pieces and make a homogenate, centrifuge the prepared 10% homogenate, and save the supernatant for later use. S6: SPSS20.0 statistical software was used for data statistics. The quantitative data that conformed to the normal distribution and had equal variance were expressed as x±s. Factorial design variance analysis was used to compare the blood gas, sperm motility, concentration, deformity rate, MDA, and SOD of rats at different simulated altitudes.

4. The experimental method for simulating the reproductive effects of rats under low-pressure hypoxia environment exposure according to claim 3, characterized in that: The rats were raised in an environment with a temperature of 21-24°C, a humidity of 50%-60%, a 12-h day and night rhythm, and free access to food and water. They were randomly divided into three groups, 20 in each group, and raised for 14 days; the plain group (local altitude) was raised in a clean room in the extravehicular animal center and given normoxic treatment, and the model group was divided into two groups, plateau model group I (4000m above sea level) and plateau model group II (6000m above sea level).

5. The experimental method for simulating the reproductive effects of rats under low-pressure hypoxia environment exposure according to claim 3, characterized in that: During the ventilation process in step S1, the pressure in the cabin needs to be kept stable.

6. The experimental method for simulating the reproductive effects of rats under low-pressure hypoxia environment exposure according to claim 3, characterized in that: In step S6, one-way analysis of variance is used to compare the means of multiple samples, and a t-test is used to compare the means of two samples.

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