Establishment method and application of chemical sensitization model for respiratory tract

Abstract

The invention discloses an establishment method and application of a model for sensitization of chemicals to respiratory tracts, and the method comprises the steps: analyzing and screening the types of main chemicals used in a region, and obtaining the types of chemicals with sensitization; various indexes of each screened sensitization chemical are analyzed and recorded; carrying out animal inhalation experiments on each chemical at intervals for a certain period; performing killing, blood sampling and dissecting on the animal after the experiment; the steps of blood, biochemical and immune index detection and the like are not limited. By establishing a chemical sensitization model, physiological, biochemical, immune and pathological indexes of respiratory tract allergy caused by dangerous chemicals are systematically analyzed, and corresponding regulation and control measures are obtained, so that the regulation and control measures are standardized and unified and can be popularized and used, the supervision level of the dangerous chemicals is practically improved, and the life health of a human body and the environmental safety are protected.

Description

Technical Field

[0001] The present invention relates to the technical field of chemical sensitization, and in particular to a method for establishing a chemical respiratory tract sensitization model and its application. Background Art

[0002] With the progress of society and the advancement of science and technology, new chemicals and materials continue to emerge. Statistics show that annual chemical production currently exceeds 400 million tons, with a value exceeding $1.3 trillion. The increasingly close relationship between chemicals and humans, while bringing enormous benefits to the national economy, also poses existing or potential risks to human health and the ecological environment. It is reported that over 80% of patients with bronchial asthma also suffer from allergic rhinitis, and 15% of adult bronchial asthma cases are believed to be related to workplace chemical exposure. However, due to the complex exposure factors in production and living environments, identifying potential causative allergens is not easy. Currently, commonly used methods for identifying allergens include bronchial provocation tests (including laboratory and on-site provocations), allergen-specific IgE testing, and skin testing.

[0003] Although my country has promulgated and implemented "GB 30000.21-2013 Specification for Classification and Labelling of Chemicals Part 21: Respiratory or Skin Sensitization," which lists the classification criteria and categories for respiratory sensitizers, this evaluation standard, due to the lack of a unified and effective respiratory allergy animal model, often leads to inconsistent results when different institutions or individuals conduct toxicological safety evaluations of the same chemical. This fails to provide useful methods and means for chemical regulators and related researchers, hindering the ability and efficiency of chemical safety prevention and control, and also impacting human health and environmental safety. Summary of the Invention

[0004] In response to the above-mentioned problems, the present invention aims to provide a method for establishing and applying a chemical-induced respiratory sensitization model, systematically analyze the physiological, biochemical, immunological and pathological indicators of chemical-induced respiratory allergies, and derive corresponding regulatory measures to achieve standardization, unification and promotion of their use, thereby protecting human life and health and environmental safety.

[0005] To achieve the above-mentioned object, the technical solution adopted by the present invention is as follows: A method for establishing a chemical sensitization model for the respiratory tract, comprising the following steps:

[0006] S1. Analyze and screen the main chemical categories used in the region to obtain the categories of chemicals that are allergenic;

[0007] S2. Analyze and record the various indicators of each screened allergenic chemical;

[0008] S3. Conduct animal inhalation tests on each chemical, with the tests conducted at intervals and for a certain period;

[0009] S4. After the experiment, the animals were killed, blood was collected, and samples were obtained through dissection.

[0010] S5. Conduct various index tests and tissue pathological examinations;

[0011] S6. Use test and inspection data as preliminary data for model building;

[0012] S7. Conduct clinical applications based on the preliminary data obtained, and conduct corresponding regulation and obtain regulatory measures;

[0013] S8. Comprehensive validation based on clinical applications and regulatory measures;

[0014] S9. Establish a respiratory sensitization model based on the comprehensive verification results.

[0015] Preferably, the indicators described in step S2 include but are not limited to physiological indicators, biochemical indicators, pathological indicators, and immune indicators.

[0016] Preferably, in step S3, the animal is placed in an experimental cage for conducting an inhalation experiment, and the experimental cage is provided with a fixing device for the animal inhalation operation.

[0017] Preferably, the fixing device is a conical fixing cylinder arranged at a distance from the inner wall of one side of the experimental cage, and the outer port of the conical fixing cylinder is connected to the outside of the experimental cage.

[0018] Preferably, operating rods are provided at intervals on the side wall of the experimental cage opposite to the conical cylinder, and a soft fixing plate corresponding to the conical fixing cylinder and capable of contacting the animal is provided at the inner end of each operating rod.

[0019] Preferably, soft fixing rods are provided at circumferential intervals near the side edges of each of the soft fixing plates, and the ends of the soft fixing rods are provided with ball heads.

[0020] An application of the establishment method in the field of non-chemical technology.

[0021] The present invention has the beneficial effects of establishing a chemical sensitization model, systematically analyzing the physiological, biochemical, immunological, and pathological indicators of respiratory allergies caused by hazardous chemicals, and developing corresponding regulatory measures to achieve standardization, unification, and widespread application, effectively improving the level of hazardous chemical supervision and protecting human health and environmental safety. Furthermore, during the operation of animal inhalation of chemicals, the present application implements restrictions on the animal's inhalation of chemicals based on the animal's activity characteristics, thereby ensuring the animal's inhalation volume and experimental results, avoiding waste and pollution of chemicals, and improving the convenience of the inhalation operation, facilitating the rapid and accurate acquisition of the established sensitization model data. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a diagram of the current direct spraying operation on experimental mice.

[0023] Figure 2 Illustration of the current spraying of experimental mice in the experimental cage.

[0024] Figure 3 This is a diagram illustrating the structure of a conical fixed cylinder arranged in an experimental cage according to the present invention.

[0025] Figure 4 The figure is a diagram of the operation process of the present invention when performing the inhalation operation on experimental mice.

[0026] Figure 5 For the present invention Figure 4 On this basis, the experimental mice were subjected to successive inhalation spray operations.

[0027] Figure 6 For the present invention Figure 4 Illustration showing the experimental mouse in the opposite direction during the operation.

[0028] Figure 7 FIG. 1 is a diagram showing an attractant adapted to a conical fixed tube according to the present invention.

[0029] Figure 8 This is a diagram illustrating the flipping operation of the experimental cage under the action of the attractant of the present invention.

[0030] Figure 9 This is a diagram showing an experimental mouse moving upward in a conical fixed cylinder during the spray inhalation operation of the present invention.

[0031] Figure 10 This is a diagram illustrating the structure of a soft fixing plate for restraining experimental mice according to the present invention.

[0032] Figure 11 For the present invention Figure 10 Enlarged diagram of the structure at point A in the middle.

[0033] Figure 12This is a diagram illustrating the structure of a soft fixing rod provided in the present invention.

[0034] Figure 13 This is a diagram showing how the soft fixing rod of the present invention further restricts experimental mice.

[0035] Figure 14 This is the oral and nasal inhalation poisoning device currently used in experiments.

[0036] In the figure: 1-spray bottle; 11-spray tube; 2-experimental cage; 21-conical fixing cylinder; 3-attractant; 31-connecting rod; 32-attractant plate; 3a-attractant food; 41-operating rod; 42-soft fixing plate; 43-soft fixing rod; 44-ball head; 5-experimental mouse. DETAILED DESCRIPTION

[0037] In order to enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0038] The World Health Organization (WHO) estimates that nearly 25% of the global disease burden is related to exposure to toxic chemicals and other environmental factors. Accidental poisonings claim nearly 355,000 lives annually. Therefore, chemical safety production and management should be further scientifically based, institutionalized, and legalized.

[0039] To protect public health, relevant laws and regulations worldwide require that chemicals undergo safety evaluations before they are marketed. Chemical-induced respiratory reactions primarily include respiratory irritation and respiratory allergy. Relatively speaking, the evaluation indicators and techniques involved in respiratory allergy are much more complex than those for respiratory irritation. Based on this, the present invention provides a method for establishing a chemical-induced respiratory sensitization model, comprising the following steps:

[0040] S1. Analyze and screen the main chemicals used in the region (such as flavors and preservatives in cosmetics; metals, metals and their salts, solvents in the industrial field; additives in food; and other chemicals such as formaldehyde, aniline and its derivatives) to obtain the categories of chemicals that are allergenic.

[0041] S2. Analyze and record the various indicators of each screened allergenic chemical, including physiological indicators, biochemical indicators, pathological indicators, immune indicators and other indicators.

[0042] S3. Perform an animal inhalation test on each chemical by allowing the animal (preferably a rat 5, but also rabbits) to inhale the chemical through the mouth and nose. The operation is preferably repeated on the 1st day, the 7th day, and the 14th day, and then again on the 28th day after a 14-day interval.

[0043] Due to the influence of animal instability, in order to achieve a more accurate inhalation volume for the animal and facilitate the inhalation operation of the animal, this application places the animal in an experimental cage 2 for an inhalation experiment. The experimental cage 2 can narrow the animal's activity range, thereby facilitating inhalation during the inhalation experiment, while ensuring sufficient inhalation volume and experimental effect.

[0044] The experimental mice 5 selected in this application are still smaller and more flexible than other large animals. In the inhalation experiment, due to the pungent smell of some chemicals, it is difficult to use them in the experiment. Figure 2 When using the nasal spray bottle 1 shown, the smell of the chemical agent and the impact of the spray will cause the experimental mouse 5 to run in the experimental cage 2, resulting in the experimental mouse 5 not being able to accurately inhale the amount and the spray (inhalation) time being longer, while also causing waste of chemicals and environmental pollution; and using Figure 14 The oral and nasal inhalation poisoning device shown requires the experimenter to manually grasp each experimental mouse and perform the inhalation test one by one. This process is lengthy for multiple experimental mice in the same batch, resulting in different inhalation times between different mice and, in turn, differences in experimental results. To address this issue, a fixture for the animal inhalation operation is provided in the experimental cage 2. During the inhalation operation, this fixture further limits the animal's range of motion, allowing the spray bottle 1 to spray directly into the oral and nasal direction of the experimental mouse 5 and be inhaled, thereby improving the experimental dosage and experimental effect while reducing chemical waste and pollution. When the inhalation test is not being conducted, the experimental mouse 5 can move normally, avoiding the problem of excessive restriction of the range of motion that may affect the normal physical immunity of the experimental mouse 5 and affect sensitization.

[0045] Specifically, such as Figure 3-4 As shown, the fixing device is a conical fixing cylinder 21 set at a distance from the inner wall of one side of the experimental cage 2. The outer end of the conical fixing cylinder 21 is connected to the outside of the experimental cage 2. At the same time, each conical fixing cylinder 21 is preferably the same size as a single experimental mouse 5 to avoid two experimental mice 5 entering a single conical fixing cylinder 21 and affecting the inhalation operation. Figure 4 As shown in FIG. 1 , when the inhalation experiment is not being conducted, the experimental mouse 5 moves in the experimental cage 2 outside the conical fixed cylinder 21, and when the inhalation experiment is required, the experimental mouse 5 moves in the experimental cage 2 outside the conical fixed cylinder 21. Figure 4 The experimental cage 2 is flipped in the direction indicated by the middle arc arrow. During the flipping process, each experimental mouse 5 enters the conical fixed cylinder 21. Since the conical fixed cylinder 21 is connected to the outside of the experimental cage 2, the experimenter inserts the spray tube 11 of the spray bottle 1 deep into the conical fixed cylinder 21. Under the restriction of the conical fixed cylinder 21 and the weight of the experimental mouse 5, the experimenter can "passively" inhale a sufficient amount of chemical reagent and quickly complete the inhalation operation in sequence ( Figure 5After the experimental cage 2 is turned over, to prevent the conical fixing tube 21 from being blocked by the ground or the experimental platform, the experimental cage 2 can be preferably lifted up or placed on the edge of the experimental platform to ensure that the conical fixing tube 21 is connected to the outside, facilitating the inhalation operation.

[0046] like Figure 6 As shown, during the process of flipping the experimental cage 2, there is a situation where the experimental mouse 5 enters the conical fixed cylinder 21 in the reverse direction, and then the reversed experimental mouse 5 cannot be inhaled. During the second single inhalation, the other experimental mice 5 that have already inhaled will have an increased inhalation volume, affecting the experimental results. Therefore, in order to solve this problem, as shown in FIG. Figure 7-8 As shown, the fixture also includes an attractant 3, comprising a connecting rod 31. At intervals, attractant plates 32 are positioned on the connecting rod 31, each of which is capable of being inserted into each of the conical fixing tubes 21. Each attractant food 3a is adhered to the attractant plates 32. Before the experimental cage 2 is flipped, each attractant plate 32, each of which is attached with attractant food 3a, is first inserted into the conical fixing tube 21. The experimental cage 2 is then flipped. During the flipping process, the scent emitted by the attractant food 3a attracts the experimental mice 5, which then adjusts their orientation. Once the experimental mice 5 enter the conical fixing tube 21, their mouths and noses are uniformly oriented toward the outside of the conical fixing tube 21, facilitating inhalation. This also prevents the experimental mice 5 from inhaling more air during a second inhalation after the experimental mice 5 reverse direction, potentially affecting the experimental results.

[0047] Since the experimental rat 5 still has better flexibility than other animals, it will inevitably "jump" upwards in the conical fixed cylinder 21 due to its own mobility when it is inhaling chemicals and is affected by the impact of the chemical spray. Figure 9 The mouse body state on the right side of the middle) will be far away from the spray tube 11, so that the sprayed chemicals are not effectively inhaled by the experimental mouse 5, and the waste and pollution of chemicals are caused by the loss of chemicals in the environment. Therefore, in order to solve this problem, Figure 10-11As shown, operating rods 41 are intermittently positioned on the side wall of the experimental cage 2 opposite the conical cylinder (preferably, the same as the connecting rod 31 of the attractor 3, so that each operating rod 41 is integrally connected to facilitate actuation of the entire operating rod 41). A soft fixing plate 42 (preferably a rubber plate, to prevent crushing or discomfort upon contact with the experimental mouse 5) is provided at the inner end of each operating rod 41, corresponding to the conical fixed cylinder 21 and capable of contacting the animal. After the experimental mouse 5 is oriented by the aforementioned attracting method, before the chemical spraying operation, the connecting rod 31 is actuated simultaneously with the multiple operating rods 41, and the soft fixing plates 42 at the ends of the operating rods 41 gradually approach the tail end of the experimental mouse 5. The weight of the connecting rod 31, operating rod 41, and soft fixing plates 42 is preferably applied to each experimental mouse 5, preventing the experimental mouse 5 from "jumping" upward within the conical fixed cylinder 21 and away from the spray tube 11, thereby ensuring that the experimental mouse 5 inhales a sufficient amount of the chemical. After the inhalation is completed, the experimental cage 2 is turned over and the operating rod 41 is reset to ensure the movement space of the experimental mouse 5.

[0048] On the basis of further restricting the experimental mouse 5 by the operating rod 41 and the soft fixing plate 42, due to the flexibility of the experimental mouse 5, when it is affected by the pungent and impact of the chemical spray, Figure 9 As shown, the mouth and nose of the experimental mouse 5 will move left and right and away from the spray tube 11, which will also affect the amount of chemicals inhaled and the experimental effect. Therefore, in order to further solve this problem, as shown in FIG. Figure 12 As shown, each of the soft fixing plates 42 is provided with soft fixing rods 43 (preferably made of rubber) along the circumferential spacing near the side, and its function is as follows: Figure 13 As shown, when the operating rod 41 is driven to make the soft fixing plate 42 approach the tail end of the experimental mouse 5, since the tail end of the conical fixing cylinder 21 is larger than the front end, the soft fixing rod 43 can be inserted into the conical fixing cylinder 21. The operating rod 41 is continued to be driven, and the soft fixing rod 43 is bent in the conical fixing cylinder 21 and penetrates into the gap between the conical fixing cylinder and the experimental mouse 5. The soft fixing rod 43 is gradually bent in the conical fixing cylinder 21, so that multiple soft fixing rods 43 together form a gradually shrinking space, and the soft fixing rod 43 can further extend to the head of the experimental mouse 5, contacting the head to limit the head deviation, thereby ensuring the inhalation of a sufficient amount of chemicals and the accuracy of the experiment during the spraying process, as well as the convenience of the inhalation operation.

[0049] In order to further avoid discomfort caused by the end of the soft fixing rod 43 moving into contact with the skin of the experimental mouse 5 during the process of the soft fixing rod 43 penetrating into the conical fixing cylinder 21, it is preferred that Figure 13As shown, the ends of the soft fixing rods 43 are each provided with ball heads 44. Compared to the flat and cornered soft fixing rods 43, the ball heads 44 have a smoother surface contact, thereby further reducing discomfort to the experimental mouse 5 when in contact with the moving rods 43, and facilitating stability of the experimental mouse 5 during the inhalation operation.

[0050] The experimental cage 2 of the present application can be changed in size according to the size of different selected animals (such as rabbits), or the experimental cage 2 can be selected to have a telescopic rod. When different experimental animals are selected, the telescopic rod constituting the experimental cage 2 can be adjusted to adjust the internal space, thereby improving the applicability of the experimental cage 2.

[0051] S4. After a certain period of inhalation experiment, the experimental mouse 5 is killed, blood is collected, and the mouse is dissected and sampled.

[0052] S5. Conduct tests including blood, biochemical, and immune indicators, and perform tissue pathological examinations.

[0053] S6. Use the test and inspection data as preliminary data (results) for building the model.

[0054] S7. Conduct clinical application based on the preliminary data (results) obtained, make corresponding adjustments and obtain regulatory measures (treatment control methods) based on clinical responses.

[0055] S8. Conduct comprehensive verification based on clinical applications and regulatory measures (treatment control methods) to obtain the optimal regulatory measures.

[0056] S9. Establish a respiratory sensitization model based on the comprehensive verification results, and promote its use in a standardized and unified manner based on the established model.

[0057] According to the method for establishing the sensitization model, it can be applied in the field of non-chemical technology.

[0058] The above shows and describes the basic principles, main features and advantages of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of protection claimed by the present invention.

Claims

1. A method for establishing a chemical-induced respiratory sensitization model, characterized in that: The following steps are involved: S1. Analyze and screen the main chemical categories used in the region to obtain the categories of chemicals that are allergenic; S2. Analyze and record the various indicators of each screened allergenic chemical; S3. Conduct animal inhalation tests on each chemical, with the tests conducted at intervals and for a certain period; S4. After the experiment, the animals were killed, blood was collected, and samples were obtained through dissection. S5. Conduct various index tests and tissue pathological examinations; S6. Use test and inspection data as preliminary data for model building; S7. Conduct clinical applications based on the preliminary data obtained, and conduct corresponding regulation and obtain regulatory measures; S8. Comprehensive validation based on clinical applications and regulatory measures; S9. Establish a respiratory sensitization model based on the comprehensive verification results.

2. The establishment method according to claim 1, characterized in that The indicators described in step S2 include but are not limited to physiological indicators, biochemical indicators, pathological indicators, and immune indicators.

3. The establishment method according to claim 2, characterized in that: In step S3, the animal is placed in an experimental cage for an inhalation experiment, wherein the experimental cage is provided with a fixing device for the animal inhalation operation.

4. The establishment method according to claim 3, characterized in that: The fixing device is a conical fixing cylinder arranged at a distance from the inner wall of one side of the experimental cage, and the outer port of the conical fixing cylinder is connected to the outside of the experimental cage.

5. The establishment method according to claim 4, characterized in that: Operating rods are arranged at intervals on the side wall of the experimental cage body opposite to the conical cylinder body. The inner end of each operating rod is provided with a soft fixing plate corresponding to the conical fixing cylinder and capable of contacting the animal.

6. The establishment method according to claim 5, characterized in that: Soft fixing rods are arranged at circumferential intervals near the side of each soft fixing plate, and ball heads are arranged at the ends of the soft fixing rods.

7. Application of the method according to claim 6 in the field of non-chemical technology.

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