Mouse cochlear radiotherapy device and radiation-induced sensorineural hearing impairment animal model

Abstract

The present invention belongs to the technical field of radiation-induced hearing impairment animal models. Disclosed are a mouse cochlear radiotherapy device and a radiation-induced sensorineural hearing impairment animal model. The mouse cochlea radiotherapy device comprises a base and a radiotherapy apparatus arranged above the base, wherein a lead plate is provided below the radiotherapy apparatus; a plurality of mounting boxes are fixed to a bottom end of the base, and a plurality of sliding grooves passing through the mounting boxes are provided on the top end of the base; a head fixing strap and two sets of limb fixing straps are respectively provided between two sliding grooves on the same side, and positioning teeth are arranged at the bottom end of each of the head fixing strap and the limb fixing straps; and toothed plates which engage with the positioning teeth are elastically provided inside the mounting boxes, and a mounting frame having an L-shaped structure is fixed to a top end of the base. The mouse cochlea radiotherapy device and the radiation-induced sensorineural hearing impairment animal model facilitate the fixation of a tested mouse, thereby improving the accuracy of radiation and the test effect.

Description

A mouse cochlear radiotherapy device and an animal model of radiation-induced sensorineural hearing loss. Technical Field

[0001] This invention belongs to the technical field of animal models of radiotherapy-induced hearing loss, specifically relating to a mouse cochlear radiotherapy device and an animal model of radiation-induced sensorineural hearing loss. Background Technology

[0002] Mouse cochlear radiotherapy is primarily used to study radiation-induced inner ear damage and its mechanisms. It investigates the effects of radiotherapy on the mouse cochlea and its auditory neural pathways, providing experimental evidence for determining appropriate radiotherapy doses and exploring the mechanisms of radiation-induced inner ear damage. A specific dose of radiation is used to irradiate the inner ear region of mice, and various test parameters, such as distortion product otoacoustic emissions (DPOAE) and auditory brainstem response (ABR), are measured within a specified time to assess changes in auditory physiological function. In addition to changes in auditory physiological function, morphological changes in the cochlea may also occur, such as the expression of the motor protein prestin in outer hair cells. During mouse cochlear radiotherapy, the mice need to be immobilized so that the radiotherapy device can be accurately aimed at the cochlea. Technical issues

[0003] Patent application number 202221039949.9 discloses a mouse restraint device, including: a cylinder for accommodating a mouse, a stop block, and a support platform, with a clamping mechanism provided on the support platform; a first opening is formed axially on the side wall of the cylinder, the first opening extends to both ends of the cylinder, one end of the cylinder is open, and a baffle is provided at the other end of the cylinder, with a second opening provided on the baffle, the first opening and the second opening being connected; the stop block includes a stop block body and a connector provided on the stop block body, the stop block body is located inside the cylinder, and the connector is slidably engaged with the first opening; the cylinder is rotatably clamped by the clamping mechanism.

[0004] Although the above-mentioned technical solution can locate the mouse, the mouse's head can still move after it is fixed, which makes it difficult for the radiotherapy device to be aimed at the mouse's cochlea, resulting in poor radiotherapy results in the mouse cochlea. Technical solutions

[0005] (1) Technical problems to be solved

[0006] To address the shortcomings of existing technologies, the present invention aims to provide a mouse cochlear radiotherapy device and an animal model of radiation-induced sensorineural hearing loss. This mouse cochlear radiotherapy device and animal model of radiation-induced sensorineural hearing loss are designed to solve the technical problem that, under existing technologies, after fixing the mouse, the mouse's head can still move, making it difficult to aim the radiotherapy device at the mouse's cochlea and resulting in poor radiotherapy effects on the mouse cochlea.

[0007] (2) Technical solution

[0008] To address the aforementioned technical problems, this invention provides a mouse cochlear radiotherapy device and an animal model of radiation-induced sensorineural hearing loss, comprising a base and a radiotherapy device mounted on the base. A lead plate is positioned below the radiotherapy device. Multiple mounting boxes are fixed to the bottom of the base, and multiple sliding grooves penetrating the mounting boxes are provided at the top of the base. A head fixation strap and two sets of limb fixation straps are respectively provided between two sliding grooves on the same side. Positioning teeth are provided at the bottom of both the head fixation strap and the limb fixation straps. A toothed plate that engages with the positioning teeth is elastically provided in the mounting box.

[0009] When using this technical solution, head and limb fixation straps are embedded in multiple grooves at the top of the base. The mouse to be tested is placed on the top of the base, and the two limb and head fixation straps are put on the mouse. By holding the protruding parts, the limb and head fixation straps are pulled down to fit on both sides of the mouse's body and head respectively. After the mouse is tightened, the spring pushes the toothed plate to engage with the positioning teeth, thus keeping the mouse fixed at the top of the base and corresponding to the radiotherapy equipment. The lead plate blocks the radiation from the radiotherapy equipment, and the radiation holes on the lead plate allow the radiation to be accurately aimed at the mouse's cochlea, which facilitates the fixation of the mouse and improves the accuracy of radiation and the experimental effect.

[0010] Preferably, an L-shaped mounting bracket is fixed to the top of the base, and load-bearing rods are fixed to both sides of the inner wall of the bracket.

[0011] Furthermore, the radiotherapy device is fixed to the top of the mounting frame, and a mounting plate connected to a lead plate is fixed to the bottom of the mounting frame. A radiation hole is opened at the center of the lead plate.

[0012] Furthermore, a spring is embedded in the mounting box, and a connecting rod is slidably inserted into the side wall of the mounting box.

[0013] Furthermore, one end of the connecting rod is fixed with a pull plate, and the other end of the connecting rod passes through a spring and is fixedly connected to the toothed plate.

[0014] Furthermore, including:

[0015] Animal selection for the experiment: The mouse strains selected included C57BL / 6J and CBA / caJ, and the rat strains included SD and Wistar.

[0016] Radiation treatment: The radiation source is a radiotherapy device, and the radiation location is the cochlea, to ensure that the main impact is on the auditory system while minimizing damage to other organs;

[0017] Control group setup: A normal control group that did not receive radiation treatment was set up to compare physiological changes before and after radiation.

[0018] Hearing function testing includes auditory brainstem response and distortion product otoacoustic emissions;

[0019] Histological analysis: including cochlear basilar membrane spreads, cochlear axial sections, scanning electron microscopy and transmission electron microscopy, and histological staining;

[0020] Molecular biology detection, including gene expression analysis, proteomics, and signaling pathway analysis.

[0021] Furthermore, the experimental animals are selected as adults or near-adults to avoid the influence of physiological changes during development on the experimental results. The sex selection needs to be determined according to the research purpose, and the influence of sex differences on radiation response needs to be considered.

[0022] Furthermore, the auditory brainstem response measures the reaction time (threshold) of brainstem neurons after sound stimulation to assess hearing sensitivity; the distortion product otoacoustic emission detects the function of outer hair cells, reflecting the functional state of the inner ear.

[0023] Furthermore, the cochlear swabs are used to observe morphological changes in hair cells, supporting cells, and nerve fibers; the scanning electron microscope and transmission electron microscope are used to observe cell ultrastructure, such as changes in mitochondria and endoplasmic reticulum, at high resolution; the histological staining includes H&E staining and immunofluorescence staining.

[0024] Furthermore, the gene expression analysis detects changes in the expression of related genes, the proteomics detects the expression levels of key proteins, and the signaling pathway analysis affects the function and survival of auditory cells through specific signaling pathways. Beneficial effects

[0025] (3) Beneficial effects

[0026] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0027] This invention utilizes multiple grooves at the top of the base to embed head and limb fixation straps. The mouse to be tested is placed on the top of the base, and the two limb and head fixation straps are fitted onto the mouse. By grasping the protruding parts, the limb and head fixation straps are pulled downwards, securing them to the sides of the mouse's body and head respectively. After tightening, a spring-driven toothed plate engages with the positioning teeth, thus fixing the mouse at the top of the base and aligning it with the radiotherapy equipment. The lead plate blocks the radiation from the equipment, and the radiation holes on the lead plate allow the radiation to be precisely aimed at the mouse's cochlea, facilitating mouse fixation and improving the accuracy and effectiveness of the radiation therapy. Attached Figure Description

[0028] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0029] Figure 1 is a schematic diagram of the structure of the present invention;

[0030] Figure 2 is a cross-sectional view of the base in this invention;

[0031] Figure 3 is an enlarged schematic diagram of the structure at point A in Figure 2 of the present invention;

[0032] Figure 4 is a schematic diagram of the hearing threshold of the present invention;

[0033] Figure 5 is a schematic diagram showing the self-comparison of the brainstem response threshold of the present invention;

[0034] Figure 6 is a schematic diagram showing the threshold changes of pure tone hearing at different frequencies before and after irradiation according to the present invention.

[0035] Figure 7 is a schematic diagram of the hair cells of the inner ear basilar membrane at the top, middle, and bottom of the inner ear according to the present invention;

[0036] Figure 8 is a schematic diagram of cochlear spiral ganglion neurons shown in the cochlear axial section of the present invention.

[0037] The markings in the attached diagram are as follows: 1. Base; 2. Mounting bracket; 3. Load-bearing rod; 4. Radiation port; 5. Radiotherapy equipment; 6. Mounting plate; 7. Lead plate; 8. Protrusion; 9. Head fixation strap; 10. Limb fixation strap; 11. Slide groove; 12. Positioning tooth; 13. Connecting rod; 14. Pull plate; 15. Mounting box; 16. Spring; 17. Toothed plate. Embodiments of the present invention

[0038] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0039] This specific embodiment is a mouse cochlear radiotherapy device and an animal model of radiation-induced sensorineural hearing loss. Its structural schematic diagram is shown in Figures 1-4. It includes a base 1 and a radiotherapy device 5 disposed on the base 1. A lead plate 7 is disposed below the radiotherapy device 5. Multiple mounting boxes 15 are fixed to the bottom end of the base 1, and multiple sliding grooves 11 penetrating the mounting boxes 15 are opened at the top end of the base 1. A head fixation strap 9 and two sets of limb fixation straps 10 are respectively disposed between two sliding grooves 11 on the same side. The bottom end of the head fixation strap 9 and the limb fixation straps 10 are provided with positioning teeth 12. A toothed plate 17 that engages with the positioning teeth 12 is elastically disposed in the mounting box 15.

[0040] An L-shaped mounting bracket 2 is fixed to the top of the base 1. Load-bearing rods 3 are fixed to both sides of the inner wall of the bracket. The radiotherapy device 5 is fixed to the top of the mounting bracket 2. A mounting plate 6 connected to a lead plate 7 is fixed to the bottom of the mounting bracket 2. A radiation hole 4 is opened at the center of the lead plate 7. A spring 16 is embedded in the mounting box 15, and a connecting rod 13 is slidably inserted into the side wall of the mounting box 15. A pull plate 14 is fixed to one end of the connecting rod 13, and the other end of the connecting rod 13 passes through the spring 16 and is fixedly connected to a toothed plate 17. The mouse to be tested is placed on the top of the base 1, and its two limbs are fixed. The limb fixation strap 10 and head fixation strap 9 are fitted onto the mouse. By holding the protrusion 8, the limb fixation strap 10 and head fixation strap 9 are pulled downwards respectively, so that the limb fixation strap 10 and head fixation strap 9 are fitted onto the sides of the mouse's body and the mouse's head respectively. After the mouse is tightened, the spring 16 pushes the toothed plate 17 to engage with the positioning tooth 12, thereby keeping the mouse fixed at the top of the base 1 and corresponding to the radiotherapy device 5. The lead plate 7 blocks the radiation from the radiotherapy device 5. At the same time, the radiation hole 4 on the lead plate 7 allows the radiation to be accurately aimed at the mouse's cochlea, which facilitates the fixation of the experimental mouse and improves the accuracy of the radiation and the experimental effect.

[0041] As shown in Figures 5-8, it includes:

[0042] Animal selection for the experiment: The mouse strains selected included C57BL / 6J and CBA / caJ, and the rat strains included SD and Wistar.

[0043] Radiation treatment: The radiation source is radiotherapy equipment 5, and the radiation location is the cochlea, to ensure that the main impact is on the auditory system while minimizing damage to other organs;

[0044] Control group setup: A normal control group that did not receive radiation treatment was set up to compare physiological changes before and after radiation.

[0045] Hearing function testing includes auditory brainstem response and distortion product otoacoustic emissions;

[0046] Histological analysis: including cochlear basilar membrane spreads, cochlear axial sections, scanning electron microscopy and transmission electron microscopy, and histological staining;

[0047] Molecular biology detection, including gene expression analysis, proteomics, and signaling pathway analysis.

[0048] The experimental animals were selected as adults or near-adults to avoid the influence of physiological changes during development on the experimental results. The sex selection should be determined according to the research purpose, and the influence of sex differences on radiation response should be considered.

[0049] The auditory brainstem response measures the reaction time (threshold) of brainstem neurons after sound stimulation to assess hearing sensitivity; the distortion product otoacoustic emission detects the function of outer hair cells, reflecting the functional state of the inner ear.

[0050] The cochlear swabs were used to observe morphological changes in hair cells, supporting cells, spiral ganglion neurons, and nerve fibers; the scanning electron microscope and transmission electron microscope were used to observe cell ultrastructure, such as changes in mitochondria and endoplasmic reticulum, at high resolution; the histological staining included H&E staining and immunofluorescence staining.

[0051] The gene expression analysis detects changes in the expression of related genes, the proteomics detects the expression levels of key proteins, and the signaling pathway analysis affects the function and survival of auditory cells through specific signaling pathways.

[0052] Working principle: When using the device of this technical solution, the mouse to be tested is placed on the top of the base 1. The two limb fixation straps 10 and the head fixation strap 9 are respectively placed on the mouse's body. By holding the protrusion 8, the limb fixation straps 10 and the head fixation strap 9 are pulled down respectively, so that the limb fixation straps 10 and the head fixation strap 9 are respectively placed on the sides of the mouse's body and the head of the mouse. After the mouse is tightened, the spring 16 pushes the toothed plate 17 to engage with the positioning teeth 12, thereby keeping the mouse fixed on the top of the base 1 and corresponding to the radiotherapy device 5. The lead plate 7 blocks the radiation from the radiotherapy device 5. At the same time, the radiation hole 4 on the lead plate 7 allows the radiation to be accurately aimed at the mouse's cochlea, which facilitates the fixation of the test mouse and improves the accuracy of radiation and the test effect. (Figure 4) Figure 5 shows the hearing thresholds corresponding to different sound frequencies on the 10th day after cochlear irradiation at different doses. Figure 6 shows the self-comparison of the auditory brainstem response threshold for click sounds before and after irradiation with a 30 Gy dose of cochlear irradiation. It can be seen from the figure that the auditory brainstem response threshold for click sounds increased significantly after 10 days of irradiation. Figure 7 shows the changes in thresholds under pure tone hearing at different frequencies before and after irradiation. The threshold also increased significantly after 10 days of irradiation. Figure 8 shows the hair cells of the apical, mid, and basal turns of the basilar membrane of the inner ear on the 10th day after cochlear irradiation (no obvious hair cell death was observed). Figure 9 shows the cochlear spiral ganglion neurons in the control group and on the 10th day after irradiation (see changes in cell morphology, cell atrophy, and deep staining of cell nuclei).

[0053] All technical features in this embodiment can be freely combined according to actual needs.

[0054] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A mouse cochlear radiotherapy device, comprising a base (1) and a radiotherapy device (5) disposed above the base (1), characterized in that, A lead plate (7) is provided below the radiotherapy device (5). Multiple mounting boxes (15) are fixed at the bottom of the base (1). Multiple sliding grooves (11) penetrating the mounting boxes (15) are provided at the top of the base (1). A head fixation strap (9) and two sets of limb fixation straps (10) are respectively provided between two sliding grooves (11) on the same side. Positioning teeth (12) are provided at the bottom of the head fixation strap (9) and the limb fixation straps (10). A toothed plate (17) that engages with the positioning teeth (12) is elastically provided in the mounting box (15).

2. The mouse cochlear radiotherapy device according to claim 1, characterized in that, The top of the base (1) is fixed with an L-shaped mounting bracket (2), and load-bearing rods (3) are fixed on both sides of the inner wall of the bracket.

3. The mouse cochlear radiotherapy device according to claim 2, characterized in that, The radiotherapy device (5) is fixed at the top of the mounting frame (2), and the bottom of the mounting frame (2) is fixed with a mounting plate (6) connected to the lead plate (7). A radiation hole (4) is opened at the center of the lead plate (7).

4. The mouse cochlear radiotherapy device according to claim 1, characterized in that, A spring (16) is embedded in the mounting box (15), and a connecting rod (13) is slidably inserted into the side wall of the mounting box (15).

5. The mouse cochlear radiotherapy device according to claim 4, characterized in that, One end of the connecting rod (13) is fixed with a pull plate (14), and the other end of the connecting rod (13) passes through a spring (16) and is fixedly connected to the toothed plate (17).

6. An animal model of radiation-induced sensorineural hearing loss, using a mouse cochlear radiotherapy device according to any one of claims 1-5, characterized in that, include: Animal selection for the experiment: The mouse strains selected included C57BL / 6J and CBA / caJ, and the rat strains included SD and Wistar. Radiation treatment: The radiation source is a radiotherapy device, and the radiation location is the cochlea, to ensure that the main impact is on the auditory system while minimizing damage to other organs; Control group setup: A normal control group that did not receive radiation treatment was set up to compare physiological changes before and after radiation. Hearing function testing includes auditory brainstem response and distortion product otoacoustic emissions; Histological analysis: including cochlear basilar membrane spreads, cochlear axial sections, scanning electron microscopy and transmission electron microscopy, and histological staining; Molecular biology detection, including gene expression analysis, proteomics, and signaling pathway analysis.

7. The animal model of radiation-induced sensorineural hearing loss according to claim 6, characterized in that, The experimental animals were selected as adults or near-adults to avoid the influence of physiological changes during development on the experimental results. The sex selection should be determined according to the research purpose, and the influence of sex differences on radiation response should be considered.

8. The animal model of radiation-induced sensorineural hearing loss according to claim 6, characterized in that, The auditory brainstem response measures the reaction time (threshold) of brainstem neurons after sound stimulation to assess hearing sensitivity; the distortion product otoacoustic emission detects the function of outer hair cells, reflecting the functional status of the middle and inner ear.

9. The animal model of radiation-induced sensorineural hearing loss according to claim 6, characterized in that, The cochlear swabs were used to observe morphological changes in hair cells, supporting cells, and nerve fibers; the scanning electron microscope and transmission electron microscope were used to observe cell ultrastructure, such as changes in mitochondria and endoplasmic reticulum, at high resolution; the histological staining included H&E staining and immunofluorescence staining.

10. The animal model of radiation-induced sensorineural hearing loss according to claim 6, characterized in that, The gene expression analysis detects changes in the expression of related genes, the proteomics detects the expression levels of key proteins, and the signaling pathway analysis affects the function and survival of auditory cells through specific signaling pathways.

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