Mouse cochlea radiotherapy device and radioactive sensorineural hearing impairment animal model

Abstract

The invention discloses a mouse cochlea radiotherapy device and a radioactive sensorineural hearing impairment animal model, and belongs to the technical field of radiotherapy hearing impairment animal models.The mouse cochlea radiotherapy device comprises a base and radiotherapy equipment arranged above the base, a lead plate is arranged below the radiotherapy equipment, a plurality of mounting boxes are fixed to the bottom end of the base, and the mounting boxes are connected with the lead plate. A mounting box is arranged on the base, a plurality of sliding grooves penetrating through the mounting box are formed in the top end of the base, a head fixing belt and two sets of limb fixing belts are arranged between the two sliding grooves on the same side, positioning teeth are arranged at the bottom ends of the head fixing belt and the limb fixing belts, and a toothed plate clamped with the positioning teeth is elastically arranged in the mounting box. According to the mouse cochlea radiotherapy device and the radioactive sensorineural hearing injury animal model, a test mouse can be conveniently fixed, and the accuracy of radioactive rays and the test effect are improved.

Description

Technical Field

[0001] The invention belongs to the technical field of radiotherapy hearing loss animal models, and specifically relates to a mouse cochlear radiotherapy device and a radiation sensorineural hearing loss animal model. Background Art

[0002] Mouse cochlear radiotherapy is mainly used to study radiation-induced inner ear damage and its mechanism. To explore the effects of radiotherapy on the mouse cochlea and its auditory nerve pathway, and to provide experimental basis for formulating reasonable radiotherapy doses in clinic and exploring the mechanism of radiation-induced inner ear damage. A specific dose of radiation is used to irradiate the mouse inner ear area, and then various indicators to be tested, such as distortion product otoacoustic emissions (DPOAE) and auditory brainstem responses (ABR), are detected within a specified time to evaluate changes in auditory physiological function. In addition to changes in auditory physiological function, changes in cochlear morphology may also occur, such as the expression of the outer hair cell motor protein prestin. When radiotherapy is performed on the mouse cochlea, the mouse needs to be fixed so that the radiotherapy device can be accurately aimed at the mouse cochlea.

[0003] The patent with application number 202221039949.9 discloses a mouse fixer, including: a cylinder for accommodating mice, a block, and a support platform, wherein a clamping mechanism is arranged on the support platform; a first opening is axially opened on the side wall of the cylinder, and the first opening extends to both ends of the cylinder, one end of the cylinder is open, and a baffle is arranged at the other end of the cylinder, and a second opening is arranged on the baffle, and the first opening is connected to the second opening; the block includes a block body and a connecting piece arranged on the block body, the block body is located in the cylinder, and the connecting piece is slidably matched with the first opening; the cylinder can be rotatably clamped in the clamping mechanism.

[0004] Although the above technical solution can position the mouse, the head of the mouse can still shake after the mouse is fixed, which makes it difficult for the radiotherapy device to aim at the cochlea of ​​the mouse, resulting in poor radiotherapy test results on the cochlea of ​​the mouse. Summary of the invention

[0005] (1) Technical issues to be solved

[0006] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a mouse cochlear radiotherapy device and an animal model of radiation sensorineural hearing loss. The mouse cochlear radiotherapy device and the animal model of radiation sensorineural hearing loss are intended to solve the technical problem that after the mouse is fixed in the prior art, the mouse's head can still shake, which makes it difficult to aim the radiotherapy device at the mouse's cochlea, resulting in poor radiotherapy test results on the mouse cochlea.

[0007] (2) Technical solution

[0008] In order to solve the above technical problems, the present invention provides a mouse cochlear radiotherapy device and an animal model of radiation sensorineural hearing loss, comprising a base and a radiotherapy device arranged above the base, a lead plate is arranged under the radiotherapy device, a plurality of mounting boxes are fixed to the bottom end of the base, and a plurality of slide grooves penetrating the mounting boxes are opened at the top of the base, a head fixing strap and two groups of limb fixing straps are respectively arranged between two of the slide grooves on the same side, positioning teeth are arranged at the bottom ends of the head fixing strap and the limb fixing strap, and a tooth plate engaged with the positioning teeth is elastically arranged in the mounting box.

[0009] When using the technical solution, a head fixing strap and a limb fixing strap are respectively embedded in a plurality of slide grooves opened at the top of the base, and the mouse to be tested is placed on the top of the base, and the two limb fixing straps and the head fixing strap are respectively put on the mouse's body, and the limb fixing straps and the head fixing strap are respectively pulled downward by holding the protrusions, so that the limb fixing straps and the head fixing straps are respectively put on both sides of the mouse's body and the mouse's head, and after the mouse is tightly put on, the spring pushes the tooth plate and the positioning tooth to engage with each other, so that the mouse is kept fixed on the top of the base and corresponds to the radiotherapy equipment. The lead plate blocks the rays of the radiotherapy equipment, and at the same time, the radiation holes on the lead plate enable the rays to be accurately aimed at the mouse's cochlea, which is convenient for fixing the experimental mouse and improves the accuracy of the radiation and the test effect.

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

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

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

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

[0014] Further, including:

[0015] Selection of experimental animals: The selected mouse strains include C57BL / 6J and CBA / caJ, and the rats include SD and Wistar strains;

[0016] Radiation treatment: The radiation source is a radiotherapy device, and the radiation location is the cochlea, to ensure that it mainly affects the auditory system and minimizes damage to other organs;

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

[0018] Hearing function test: including auditory brainstem response and distortion product otoacoustic emission;

[0019] Histomorphological analysis: including cochlear basilar membrane flat mounts, cochlear midaxis sections, scanning electron microscopy, transmission electron microscopy, and histological staining;

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

[0021] Furthermore, the experimental animals are selected from adult or nearly adult animals to avoid the influence of physiological changes during development on the experimental results. The gender selection should be determined according to the research purpose, and the influence of gender differences on radiation response should be considered.

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

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

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

[0025] (3) Beneficial effects

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] The present invention has a plurality of slide grooves provided at the top of the base, each of which is provided with a head fixing belt and a limb fixing belt. A mouse to be tested is placed at the top of the base, and two limb fixing belts and a head fixing belt are respectively put on the mouse. The limb fixing belts and the head fixing belt are respectively pulled downward by holding the raised parts, so that the limb fixing belts and the head fixing belt are respectively put on both sides of the mouse's body and the mouse's head. After the mouse is tightly put on, a spring pushes the tooth plate and the positioning tooth to engage with each other, so that the mouse is kept fixed at the top of the base and corresponds to the radiotherapy equipment. The lead plate blocks the radiation of the radiotherapy equipment. At the same time, the radiation hole on the lead plate enables the radiation to be accurately aimed at the cochlea of ​​the mouse, which is convenient for fixing the test mouse and improves the accuracy of the radiation and the test effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For those skilled in the art, other drawings can be obtained based on these drawings without paying any creative work.

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

[0030] Figure 2 It is a schematic cross-sectional structural diagram of the base in the present invention;

[0031] Figure 3 For the present invention Figure 2 A schematic diagram of the structure enlargement in the middle;

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

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

[0034] Figure 6 It is a schematic diagram of the change of pure tone hearing thresholds of different frequencies before and after irradiation of the present invention;

[0035] Figure 7 Schematic diagram of the apical, middle and basal rotation of the hair cells of the inner ear basilar membrane of the present invention;

[0036] Figure 8 This is a schematic diagram of the neuron cells of the spiral ganglion of the cochlea shown in the cochlear mid-axis section of the present invention.

[0037] The marks in the attached drawings are: 1. base; 2. mounting frame; 3. load-bearing rod; 4. radiation hole; 5. radiotherapy equipment; 6. mounting plate; 7. lead plate; 8. raised part; 9. head fixing belt; 10. limb fixing belt; 11. slide groove; 12. positioning tooth; 13. connecting rod; 14. pull plate; 15. mounting box; 16. spring; 17. tooth plate. DETAILED DESCRIPTION

[0038] 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 described embodiments 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.

[0039] This specific embodiment is a mouse cochlear radiotherapy device and a radiation-induced sensorineural hearing loss animal model, and its structural schematic diagram is shown in FIG. Figure 1-Figure 4As shown, it includes a base 1 and a radiotherapy device 5 arranged above the base 1, a lead plate 7 is arranged below the radiotherapy device 5, a plurality of mounting boxes 15 are fixed to the bottom end of the base 1, and a plurality of slide grooves 11 penetrating the mounting boxes 15 are opened at the top of the base 1, a head fixing belt 9 and two groups of limb fixing belts 10 are respectively arranged between the two slide grooves 11 on the same side, the bottom ends of the head fixing belt 9 and the limb fixing belt 10 are both provided with positioning teeth 12, and a tooth plate 17 engaged with the positioning teeth 12 is elastically arranged in the mounting box 15.

[0040] The top of the base 1 is fixed with an L-shaped mounting frame 2, and the inner walls of the frame are fixed with load-bearing rods 3 on both sides. The radiotherapy equipment 5 is fixed on the top of the mounting frame 2, and the bottom of the mounting frame 2 is fixed with a mounting plate 6 connected to a lead plate 7, and a radiation hole 4 is provided 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 the tooth plate 17. The mouse to be tested is placed on the top of the base 1, and the two limbs are fixed respectively. The belt 10 and the head fixing belt 9 are put on the mouse, and the raised part 8 is held by hand to pull down the limb fixing belt 10 and the head fixing belt 9 respectively, so that the limb fixing belt 10 and the head fixing belt 9 are respectively put on the two sides of the mouse's body and the mouse's head, and after the mouse is tightly put on, the spring 16 pushes the tooth plate 17 and the positioning tooth 12 to engage with each other, so that the mouse is kept fixed on the top of the base 1 and corresponds to the radiotherapy equipment 5. The lead plate 7 blocks the rays of the radiotherapy equipment 5. At the same time, the radiation hole 4 on the lead plate 7 enables the rays to be accurately aimed at the mouse's cochlea, which is convenient for fixing the experimental mouse and improves the accuracy of the radiation and the experimental effect.

[0041] like Figure 5-Figure 8 As shown, including:

[0042] Selection of experimental animals: The selected mouse strains include C57BL / 6J and CBA / caJ, and the rats include SD and Wistar strains;

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

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

[0045] Hearing function test: including auditory brainstem response and distortion product otoacoustic emission;

[0046] Histomorphological analysis: including cochlear basilar membrane flat mounts, cochlear midaxis sections, scanning electron microscopy, transmission electron microscopy, and histological staining;

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

[0048] The experimental animals are selected from adults or nearly adults to avoid the influence of physiological changes during development on the experimental results. The gender selection should be determined according to the research purpose, and the influence of gender differences on radiation response should be considered.

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

[0050] The cochlear flat mounts are used to observe the morphological changes of hair cells, supporting cells, spiral ganglion neurons and nerve fibers; the scanning electron microscope and transmission electron microscope utilize high resolution to observe the ultrastructure of cells, such as changes in mitochondria and endoplasmic reticulum; and the histological staining includes H&E staining and immunofluorescence staining.

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

[0052] Working principle: When using the device of the present technical solution, place the mouse to be tested on the top of the base 1, and respectively put two limb fixing belts 10 and the head fixing belt 9 on the mouse's body, and hold the raised part 8 to pull down the limb fixing belt 10 and the head fixing belt 9 respectively, so that the limb fixing belt 10 and the head fixing belt 9 are respectively put on both sides of the mouse's body and the mouse's head, and after the mouse is tightly put on, the spring 16 pushes the tooth plate 17 and the positioning tooth 12 to engage with each other, so that the mouse is kept fixed on the top of the base 1 and corresponds to the radiotherapy equipment 5, the lead plate 7 blocks the radiation of the radiotherapy equipment 5, and at the same time, the radiation hole 4 on the lead plate 7 enables the radiation to be accurately aimed at the mouse's cochlea, which is convenient for fixing the experimental mouse, improving the accuracy of the radiation and the test effect. Figure 4 This is a diagram showing the hearing thresholds corresponding to different sound frequencies on the 10th day after cochlear radiation at different doses. Figure 5 This is a schematic diagram of the comparison of the threshold of auditory brainstem response to click sound before and after cochlear irradiation with a dose of 30 Gy. It can be concluded from the figure that the threshold of auditory brainstem response to click sound increased significantly ten days after irradiation. Figure 6 This is a diagram showing the change in thresholds for pure tone hearing at different frequencies before and after irradiation. Ten days after irradiation, the threshold also showed a significant increase. Figure 7 This is a schematic diagram of the apical, middle, and basal hair cells of the inner ear basilar membrane on the 10th day after cochlear irradiation (no obvious hair cell death was observed). Figure 8Schematic diagram of the spiral ganglion neurons in the cochlear mid-axis sections of the control group and the 10th day after irradiation (see changes in cell morphology, cell atrophy, and dark staining of the nucleus).

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

[0054] Finally, it should be noted that the above 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 aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A mouse cochlear radiotherapy device, comprising a base (1) and a radiotherapy device (5) arranged above the base (1), characterized in that: A lead plate (7) is arranged below the radiotherapy device (5), a plurality of installation boxes (15) are fixed to the bottom end of the base (1), and a plurality of slide grooves (11) penetrating the installation boxes (15) are arranged at the top end of the base (1), a head fixing belt (9) and two groups of limb fixing belts (10) are arranged between two of the slide grooves (11) on the same side, positioning teeth (12) are arranged at the bottom ends of the head fixing belt (9) and the limb fixing belt (10), and a tooth plate (17) which is engaged with the positioning teeth (12) is elastically arranged in the installation box (15).

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

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

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

5. A mouse cochlear radiotherapy device according to claim 4, characterized in that: 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 a spring (16) and is fixedly connected to a toothed plate (17).

6. A radiation-induced sensorineural hearing loss animal model, using a mouse cochlear radiotherapy device according to any one of claims 1 to 5, characterized in that: include: Selection of experimental animals: The selected mouse strains include C57BL / 6J and CBA / caJ, and the rats include SD and Wistar strains; Radiation treatment: The radiation source is a radiotherapy device, and the radiation location is the cochlea, to ensure that it mainly affects the auditory system and minimizes damage to other organs; Control group setting: A normal control group that did not receive radiation treatment was set up to compare the physiological changes before and after radiation; Hearing function test: including auditory brainstem response and distortion product otoacoustic emission; Histomorphological analysis: including cochlear basilar membrane flat mounts, cochlear midaxis sections, scanning electron microscopy, transmission electron microscopy, and histological staining; Molecular biology testing; 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 are selected from adults or nearly adults to avoid the influence of physiological changes during development on the experimental results. The gender selection should be determined according to the research purpose, and the influence of gender 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 evaluate hearing sensitivity; the distortion product otoacoustic emission detects the function of outer hair cells and reflects the functional status of the middle ear and inner ear.

9. The animal model of radiation-induced sensorineural hearing loss according to claim 6, characterized in that: The cochlear flat mount is used to observe the morphological changes of hair cells, supporting cells and nerve fibers; the scanning electron microscope and transmission electron microscope use high resolution to observe the ultrastructure of cells, such as changes in mitochondria and endoplasmic reticulum; and the histological staining includes 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 expression changes of related genes, the proteomics detects expression levels of key proteins, and the signal pathway analysis affects the function and survival of auditory cells through specific signal pathways.

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