Construction method of scoliosis rat model

Abstract

The invention belongs to the technical field of biology, and particularly relates to a scoliosis rat model construction method which comprises the following steps: S1, obtaining an experimental animal, removing the forelimb and the tail of the experimental animal, enabling the gravity direction of the experimental animal to be parallel to the spine, and obtaining a biped animal; s2, performing excision treatment on a deep paravertebral muscle group on one side of the biped animal to obtain an experimental animal with muscle mechanics imbalance; and S3, performing exercise load stimulation treatment on the experimental animal with muscle mechanics imbalance to obtain the scoliosis animal model. According to the method disclosed by the invention, the scoliosis animal model capable of highly simulating human disease characteristics is constructed by combining axial gravity load under a two-foot standing posture, structural and mechanical unbalance caused by directional unilateral deep paravertebral muscle excision and regular movement stimulation; the method can accurately and controllably induce scoliosis in a specific direction opposite to a muscle resection side, and has a wide application prospect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and particularly relates to a method for constructing a scoliosis rat model. BACKGROUND

[0002] Currently, the methods for constructing a scoliosis animal model mainly include vertebral surgery induction (such as asymmetric vertebral fixation), chemical intervention (such as paravertebral injection of bisphosphonate or relaxin), and the use of specific genetically modified animals (such as Ptk7 However, there are limitations in the degree of agreement with the natural course of human idiopathic scoliosis: surgery and chemical methods belong to acute and exogenous injury, which are difficult to accurately simulate the progressive pathogenesis of unknown etiology; and the genetic model is limited by species differences, with poor controllability of the scoliosis phenotype, long experimental period, and high cost.

[0003] Therefore, it is of great significance to develop an animal model that can stably control the direction of scoliosis, has a high success rate of modeling, and can better simulate the mechanical environment of the human spine under the action of gravity. SUMMARY

[0004] The present application aims to at least partially solve at least one of the technical problems existing in the prior art. To this end, the present application provides a method for constructing a scoliosis rat model. The method of the present application combines axial gravity load in a bipedal standing posture, structural mechanical imbalance caused by directional unilateral deep paravertebral muscle resection, and regular movement stimulation, thereby constructing an animal model of scoliosis that can highly simulate the characteristics of human disease. The method can accurately and controllably induce a specific direction of scoliosis opposite to the muscle resection side, realizing a fundamental change in the direction of modeling from "random" to "directional", and providing an ideal research platform for in-depth exploration of the pathogenesis of scoliosis and the development of new treatment strategies.

[0005] In a first aspect, the present application provides a method for constructing a scoliosis animal model. According to an embodiment of the present application, the method comprises the following steps: S1: obtaining an experimental animal, removing the forelimbs and tail of the experimental animal, making the gravity direction of the experimental animal parallel to the spine, and obtaining a bipedal animal; S2: performing resection treatment on a deep paraspinal muscle group on one side of the bipedal animal, and obtaining a muscle mechanics imbalance experimental animal; S3: performing motion load stimulation treatment on the muscle mechanics imbalance experimental animal, and obtaining the scoliosis animal model; wherein the deep paraspinal muscle group on one side includes the multifidus muscle, the rotator muscle, and the semispinal muscle; and the experimental animal is a quadruped mammal. According to the method of the embodiment of the present application, the three steps of "bipedal model establishment", "directional unilateral deep paraspinal muscle group resection", and "postoperative motion load stimulation" are organically integrated, and then a scoliosis animal model that can highly simulate the characteristics of human diseases is constructed, which completely overcomes the defects that the scoliosis direction is random and unpredictable in the traditional modeling method, has a high success rate of 100%, shows excellent stability and repeatability, and has a wide application prospect.

[0006] According to an embodiment of the present application, the method can have the following technical features: According to an embodiment of the present application, the resection treatment is located at the thoracic vertebra segment.

[0007] According to an embodiment of the present application, when the quadruped mammal is a rodent, the thoracic vertebra segment is the T7-T11 vertebra level.

[0008] According to an embodiment of the present application, the motion load stimulation treatment is treadmill training.

[0009] According to an embodiment of the present application, the animal is a rodent.

[0010] According to an embodiment of the present application, the rodent is a rat.

[0011] According to an embodiment of the present application, the experimental animal is a juvenile experimental animal in a rapid bone growth period.

[0012] According to an embodiment of the present application, the juvenile period is from weaning to sexual maturity.

[0013] In a second aspect, the present application provides a scoliosis animal model. According to an embodiment of the present application, the scoliosis animal model is constructed by the method of the first aspect.

[0014] Those skilled in the art can understand that the features and advantages described above for the method for constructing a scoliosis animal model are also applicable to the scoliosis animal model, and will not be repeated here.

[0015] According to an embodiment of the present application, the scoliosis animal model can further have the following technical features: According to an embodiment of the present application, the scoliosis animal model exhibits a scoliosis on the side opposite to the side of the deep paraspinal muscle group resection.

[0016] According to an embodiment of the present application, the Cobb angle of the scoliosis is >10 degrees.

[0017] In a third aspect of the present application, the present application provides the use of the method of the first aspect, the scoliosis animal model of the second aspect in the study of the pathogenesis of scoliosis, the screening or evaluation of drugs for treating scoliosis.

[0018] It can be understood by those skilled in the art that the features and advantages described above for the method of constructing a scoliosis animal model, the scoliosis animal model are also applicable to the application, which will not be repeated here.

[0019] Additional aspects and advantages of the present application will be in part apparent and in part pointed out in the description which follows, and by reference to the drawings. BRIEF DESCRIPTION OF DRAWINGS

[0020] The above and / or additional aspects and advantages of the present application will become apparent and be readily appreciated from the description of the embodiments, taken in conjunction with the following drawings in which: Figure 1 Figure 1 is a diagram of the anatomical structure of the paraspinal muscles of a rat in Example 1 of the present application, wherein A is an indication diagram of the superficial dorsal latissimus muscle, B is an indication diagram of the middle layer of the rat's spinal muscle, the longest muscle, and the iliocostal muscle, and C is an indication diagram of the deep paraspinal muscle (multisplit muscle, convoluted muscle, and semispinal muscle); Figure 2 Figure 2 is a diagram of the surgical procedure for constructing a double-foot model in Example 1 of the present application, wherein A-L are respectively a step-by-step diagram for anesthetizing a rat with isoflurane, a step-by-step diagram for cutting the skin of a rat, a step-by-step diagram for finding the blood supply artery of the forelimb of a rat, a step-by-step diagram for ligating the blood supply artery of the forelimb of a rat, a step-by-step diagram for cutting off the forelimb of a rat at the scapula, a step-by-step diagram for cutting off the forelimb of a rat on the same side, a step-by-step diagram for suturing the wound, a step-by-step diagram for cutting off the forelimb of a rat on the other side by the same method, a step-by-step diagram for ligating the blood vessels of the tail of a rat, a step-by-step diagram for cutting off the tail, a step-by-step diagram for postoperative anesthesia recovery of a rat at 37°C, and a photograph of the result of the completion of the construction of the double-foot model; Figure 3Figure for the surgical procedure of the left deep paraspinal muscle resection group / control group A in Example 1 of the present application, wherein A~E are respectively the surgical procedure practice figure of the control group A rats (skin preparation, disinfection, start of operation, exposure of the left deep paraspinal muscle, suture of the wound), and F~M are respectively the surgical procedure practice figure of the left deep paraspinal muscle resection group rats (skin preparation, disinfection, start of operation, exposure of the left deep paraspinal muscle, resection of the left deep paraspinal muscle, resection of the left deep paraspinal muscle, the resected left deep paraspinal muscle, suture of the wound); Figure 4 Figure for the surgical procedure of the right deep paraspinal muscle resection group / control group B in Example 1 of the present application, wherein A~E are respectively the surgical procedure practice figure of the control group B rats (skin preparation, disinfection, start of operation, exposure of the right deep paraspinal muscle, suture of the wound), and F~M are respectively the surgical procedure practice figure of the right deep paraspinal muscle resection group rats (skin preparation, disinfection, start of operation, exposure of the right deep paraspinal muscle, resection of the right deep paraspinal muscle, resection of the right deep paraspinal muscle, the resected right deep paraspinal muscle, suture of the wound); Figure 5 Figure for the running training of the control group A / B, left / right deep paraspinal muscle resection group rats in Example 1 of the present application; Figure 6 Figure for the CT examination results of the left deep paraspinal muscle resection group / control group A in Example 1 of the present application, wherein A is the modeling procedure figure of the scoliosis double-foot rat model, B is the experimental design schematic diagram of the intervention time point of the control group A, C is the experimental design schematic diagram of the intervention time point of the left deep paraspinal muscle resection group, D is the CT three-dimensional reconstruction result figure of the control group A rats (10 weeks old), E is the CT three-dimensional reconstruction result figure of the left deep paraspinal muscle resection group rats (10 weeks old), F is the CT three-dimensional reconstruction result figure of the control group A rats (14 weeks old), and G is the CT three-dimensional reconstruction result figure of the left deep paraspinal muscle resection group rats (14 weeks old); Figure 7Figures for the statistical results of the phenotype data and the surgical trace confirmation of each group of rats in Example 1 of the present application (left deep paraspinal muscle resection group / control group A), wherein A is a Cobb angle statistical result graph of the left deep paraspinal muscle resection group / control group A rats (10 weeks old), B is a Cobb angle statistical result graph of the left deep paraspinal muscle resection group / control group A rats (14 weeks old), C is a scoliosis rate statistical result graph of the left deep paraspinal muscle resection group / control group A rats (10 weeks old), D is a scoliosis rate statistical result graph of the left deep paraspinal muscle resection group / control group A rats (14 weeks old), E is a kyphosis angle statistical result graph of the left deep paraspinal muscle resection group / control group A rats (10 weeks old), F is a kyphosis angle statistical result graph of the left deep paraspinal muscle resection group / control group A rats (14 weeks old), G is a thoracic cross diameter / thoracic diameter statistical result graph of the left deep paraspinal muscle resection group / control group A rats (10 weeks old), H is a thoracic cross diameter / thoracic diameter statistical result graph of the left deep paraspinal muscle resection group / control group A rats (14 weeks old), I is an observation graph of the left sham operation position of the control group A rats verified by MRI (16 weeks old), and J is an observation graph of the left sham operation position of the left deep paraspinal muscle resection group rats verified by MRI (16 weeks old); Figure 8 Figures for the statistical results of the phenotype data and the surgical trace confirmation of each group of rats in Example 1 of the present application (right deep paraspinal muscle resection group / control group B), wherein A is a CT three-dimensional reconstruction result graph of the control group B rats (10 weeks old), B is a CT three-dimensional reconstruction result graph of the right deep paraspinal muscle resection group (10 weeks old), C is a Cobb angle statistical result graph of the right deep paraspinal muscle resection group / control group B rats (10 weeks old), D is a scoliosis rate statistical result graph of the right deep paraspinal muscle resection group / control group B rats (10 weeks old), E is a kyphosis angle statistical result graph of the right deep paraspinal muscle resection group / control group B rats (10 weeks old), F is a thoracic cross diameter / thoracic diameter statistical result graph of the right deep paraspinal muscle resection group / control group B rats (10 weeks old), G is an observation graph of the left sham operation position of the control group B rats verified by MRI (16 weeks old), and H is an observation graph of the left sham operation position of the right deep paraspinal muscle resection group rats verified by MRI (16 weeks old). DETAILED DESCRIPTION

[0021] Embodiments of the present application will be described in detail below. The embodiments described below are exemplary and are intended to explain the present application, but are not to be understood as limiting the present application.

[0022] It should be noted that the terms "first", "second" are used only for descriptive purpose, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the technical features indicated. Therefore, the features defined with "first", "second" can explicitly or implicitly include one or more of the features. Further, in the description of the present application, unless otherwise specified, the meaning of "a plurality of" is two or more.

[0023] The endpoints of the ranges and any values disclosed herein are not limited to the precise values stated. The ranges or values should be construed to be roughly about the ranges or values. For ranges, the endpoints are included as well as intervening points which are included in the ranges. For values, the values are included as well as intervening values which are included in the ranges.

[0024] In the present text, the terms "comprising" or "including" are open-ended expressions that are to be construed to mean including, but not limited to, the recited members.

[0025] In the present text, the terms "optionally", "optional" or "optional" generally mean that the event or condition subsequently described can or can not occur, and the description includes instances where the event or condition occurs and instances where it does not.

[0026] Method The present application proposes a method for constructing a scoliosis animal model. According to an embodiment of the present application, the method comprises the following steps: S1: obtaining an experimental animal, removing the forelimbs and tail of the experimental animal, making the gravity direction parallel to the spine, and obtaining a bipedal animal; S2: performing resection treatment on one side of the paravertebral muscle group of the bipedal animal, and obtaining a muscle mechanical imbalance experimental animal; S3: performing motion load stimulation treatment on the muscle mechanical imbalance experimental animal, and obtaining the scoliosis animal model; wherein the unilateral paravertebral muscle group includes the multifidus muscle, the rotator muscle and the semispinal muscle; and the experimental animal is a quadruped mammal. According to the method of the embodiment of the present application, by organically integrating the three steps of "biped model establishment", "directional unilateral deep paravertebral muscle resection" and "postoperative motion load stimulation", a scoliosis animal model capable of highly simulating human disease characteristics is obtained, and the defects of random and unpredictable scoliosis direction in the traditional modeling method are completely overcome. The success rate of modeling is as high as 100%, which shows excellent stability and repeatability, and has a wide application prospect.

[0027] In the present text, the term "one-side deep paraspinal muscle group" refers to a specific set of muscles located in the deep layer of one side (left or right) of the spine of the experimental animal, mainly functioning to maintain the stability of the spine and control the rotation between the vertebral bodies, including the three parts of multifidus, rotator and semispinalis, which are intertwined with each other and difficult to distinguish, and are collectively referred to as one-side deep paraspinal muscle group and are removed together.

[0028] In the present text, the term "quadruped mammal" refers to mammals that bear and move with the two pairs of fore and hind limbs in the natural physiological state, including but not limited to rodents (such as rats, mice), rabbits (such as domestic rabbits), and even larger mammals (such as small pigs), whose spines are homologous to humans in evolution, and the bipedal model constructed by amputating the forelimbs and tail can effectively simulate the unique biomechanical environment of the human body in which the gravity axis is parallel to the spine in an upright posture, therefore, all quadruped mammals belong to the protection scope of the present application.

[0029] It should be noted that before the exercise load stimulation treatment in step S3 is performed, the experimental animal must be waited for the surgical incision in step S2 to heal completely, and this recovery period is crucial for preventing exercise-induced wound infection and rupture, and ensuring that the experimental animal can participate in training in a relatively normal physiological state, which is a key prerequisite for successful model construction and animal welfare; for example, the recovery period is usually about 3 days, and the specific judgment criteria can include: the surgical incision is completely closed, there is no sign of infection such as redness and swelling, and the animal's mental state and basic activity have returned to normal; after the above healing conditions are met, the exercise load stimulation treatment can be started.

[0030] According to the embodiment of the present application, the resection treatment is located at the thoracic vertebra segment. Thus, by locating the resection treatment at the thoracic vertebra segment in combination with the precise positioning of the "one-side deep paraspinal muscle group", the success rate of constructing the animal model of scoliosis is further improved.

[0031] According to the embodiment of the present application, when the quadruped mammal is a rodent, the thoracic vertebra segment is the T7 to T11 vertebral level. Thus, when the experimental animal is a rodent (rat, mouse, etc.), the resection treatment is located at the T7 to T11 vertebral level of the thoracic vertebra segment in combination with the precise positioning of the "one-side deep paraspinal muscle group", which can not only ensure the induction of stable and typical scoliosis deformity, but also effectively avoid the instability of the model or the atypical deformity caused by too large or too small range.

[0032] In the present text, the term "T7 to T11 vertebral level" refers to the continuous region of the 7th thoracic vertebra to the 11th thoracic vertebra and the corresponding spinal segment structure and associated tissues of the experimental animal when the experimental animal is a rodent.

[0033] According to an embodiment of the present application, the motion load stimulation treatment is treadmill training. Thus, by using treadmill training as the motion load stimulation (simulating the motion load in normal human life), controllable, quantitative and repeatable axial load can be applied to the experimental animal, and such regular load can actively and efficiently accelerate the progression and solidification of the scoliosis deformity, thereby significantly shortening the modeling period. Exemplarily, a specific treadmill training scheme can be as follows: a starting stage (e.g., the first week after surgery): the experimental animal is trained at a slow speed of about 4 m / min for adaptability, with a daily duration of about 10 min; a progressive stage (e.g., the second week and thereafter): if the experimental animal has good balance ability, the speed is increased to about 6 m / min, the daily duration is gradually increased to about 15-20 min, and finally stabilized at about 30 min; the principle of dynamic adjustment is adhered to during the period, that is, if the experimental animal has movement disorders, the training intensity is immediately reduced to about 3 m / min for 10 min per day, and the subsequent scheme is flexibly adjusted according to the recovery condition thereof.

[0034] According to an embodiment of the present application, the animal is a rodent. Thus, such experimental animal has the outstanding advantages of clear genetic background, short breeding cycle and low feeding cost, and is very suitable for the method for constructing a scoliosis animal model according to the present application.

[0035] In this context, the term "rodent" refers to mammals (quadrupeds) belonging to the order Rodentia, which have high applicability as experimental animals in the method according to the present application. Exemplarily, the rodent includes but is not limited to rats, mice, guinea pigs, hamsters, etc., all of which are within the protection scope of the present application.

[0036] According to an embodiment of the present application, the rodent is a rat. Thus, the method for constructing a scoliosis animal model according to the present application is particularly suitable for rats.

[0037] According to an embodiment of the present application, the experimental animal is a juvenile experimental animal in a rapid bone growth period. Thus, by selecting to intervene when the experimental animal is in a rapid bone growth period, the biological characteristics of strong plasticity of the spine at this stage are ingeniously utilized, and mechanical interference is applied at this stage, so that the spine is more likely to respond to external intervention and remodel, thereby greatly improving the success rate and efficiency of modeling, ensuring that the deformity can be effectively induced and fixed, and thereby obtaining a more stable scoliosis animal model.

[0038] According to an embodiment of the present application, the juvenile stage is from weaning to sexual maturity. Thus, when the method of the present application is implemented in different species (various quadruped mammals), a clear, universal and operable biological time window criterion is provided, which is independent of the specific age or month, making the application of the method more flexible and adaptable.

[0039] Illustratively, when the experimental animal is selected as a rodent (such as a rat, a mouse, etc.), an individual with an age of 3-11 weeks can be selected to start the construction of the scoliosis animal model of the present application.

[0040] It should be noted that the aforementioned juvenile experimental animal, the individual with an age of 3-11 weeks, etc. all refer to the initial age of the experimental animal when the experimental animal is obtained in step S1 of the method of the present application and the subsequent operation of constructing a biped model (obtaining the biped animal) is performed.

[0041] According to an embodiment of the present application, the scoliosis animal model obtained by the method exhibits a scoliosis opposite to the side of the deep paraspinal muscle group resection. Thus, the scoliosis animal model obtained by the method has a stable corresponding relationship of "resection side opposite to bending side", rather than the random scoliosis conversion of the traditional model, and thus provides a research tool for studying the asymmetric pathological mechanism of scoliosis in a specific direction, the effect of drug treatment, the specific correction effect of orthopedic devices, etc. (such as the difference in bilateral bone growth of vertebral bodies, the asymmetric stress response of ligaments and intervertebral discs, etc.).

[0042] According to an embodiment of the present application, the scoliosis animal model obtained by the method has a Cobb angle of scoliosis >10 degrees. Thus, this criterion provides an objective, quantifiable and clinically diagnostic criterion for the scoliosis animal model obtained by the method. In medicine, a Cobb angle greater than 10 degrees is generally recognized as the diagnostic threshold for scoliosis. The scoliosis animal model obtained by the method of the present application can stably reach and exceed this standard, which confirms the success and effectiveness of the model construction from a quantitative point of view, and ensures the definiteness of the pathological state being studied. This not only eliminates the interference of physiological curvature, so that the scoliosis animal model can be reliably used to simulate pathological scoliosis, but also provides an accurate and measurable evaluation baseline for the correction effect of subsequent therapeutic interventions (such as drugs and devices), significantly improving the scientificity and reliability of related medical research.

[0043] It should be noted that the scoliosis animal model obtained by the method of the present application will be throughout the whole research and development chain from early screening to mid-term optimization to preclinical final evaluation of therapeutic drugs. The core features of controllable direction, stable phenotype and high clinical relevance make it an indispensable and powerful standardized tool for scoliosis research.

[0044] Animal model of scoliosis The present application provides an animal model of scoliosis. According to an embodiment of the present application, the animal model of scoliosis is obtained by the aforementioned method.

[0045] It is understood by those skilled in the art that the features and advantages described above for the method of constructing an animal model of scoliosis also apply to the animal model of scoliosis, which will not be repeated here.

[0046] According to an embodiment of the present application, the animal model of scoliosis exhibits scoliosis opposite to the side of the deep paraspinal muscle group resection.

[0047] According to an embodiment of the present application, the Cobb angle of the scoliosis is >10 degrees.

[0048] Applications The present application provides the aforementioned method, the aforementioned animal model of scoliosis for use in the study of the pathogenesis of scoliosis, and the screening or evaluation of therapeutic drugs for scoliosis.

[0049] It is understood by those skilled in the art that the features and advantages described above for the method of constructing an animal model of scoliosis, the animal model of scoliosis also apply to the application, which will not be repeated here.

[0050] The method of constructing an animal model of scoliosis, the animal model of scoliosis, and the application of the method and the animal model in the screening or evaluation of therapeutic drugs for scoliosis, are embodied in the following aspects: (1) as a standardized screening tool, for the preliminary discovery of candidate drugs: in the early stage of drug discovery, screening out substances with potential activity for inhibiting or correcting scoliosis from a large number of candidate compounds or biological agents; (2) as a precise evaluation platform, for in-depth evaluation of efficacy and mechanism: conducting comprehensive pharmacodynamics, dose response and mechanism studies on lead compounds that have shown therapeutic potential; (3) as a bridge of translational medicine, to improve the success rate of clinical prediction: before the drug enters human clinical trials, final preclinical effectiveness verification is carried out.

[0051] The solutions of the present application will be explained in connection with the embodiments. Those skilled in the art will understand that the following embodiments are only used to illustrate the present application, and should not be regarded as limiting the scope of the present application. If the specific technology or condition is not specified in the embodiments, it is carried out according to the technology or condition described in the literature in the art or according to the product instruction. If the reagent or instrument is not specified by the manufacturer, it is a conventional product that can be obtained by purchase.

[0052] Example 1: Construction of a rat model of scoliosis 1. Preparation of experimental animals Experimental animals: 3-week-old weaned SD rats (purchased from Sibeifu (Beijing) Biotechnology Co., Ltd.) were prepared, a total of 28, and were used for subsequent experiments after adaptive feeding for 3 days.

[0053] 2. Construction of scoliosis rat model The experimental animals were randomly divided into 4 groups, and the overview of the experimental animal grouping and treatment steps is shown in Table 1.

[0054] Table 1 Overview of experimental animal grouping and treatment steps

[0055] In Table 1, “constructing a double-limb model” refers to cutting off the bilateral forelimbs (cut to the scapula) and tail of the rat; “removing the left deep paraspinal muscle” refers to removing the deep paraspinal muscle (including the multifidus muscle, rotator muscle, and semispinal muscle) on the left side of the spine of the left deep paraspinal muscle removal group rat, with the removal range being the thoracic 7-thoracic 11 vertebra (T7-T11) level; “left side sham operation (incision without removal)” refers to making an incision on the left side of the control group A rat with the same size as the treatment described in “removing the left deep paraspinal muscle”, and bluntly separating to the deep paraspinal muscle, but not removing; “removing the right deep paraspinal muscle” refers to removing the deep paraspinal muscle (including the multifidus muscle, rotator muscle, and semispinal muscle) on the right side of the spine of the right deep paraspinal muscle removal group rat, with the removal range being the thoracic 7-thoracic 11 vertebra (T7-T11) level; “right side sham operation (incision without removal)” refers to making an incision on the right side of the control group B rat with the same size as the treatment described in “removing the right deep paraspinal muscle”, and bluntly separating to the deep paraspinal muscle, but not removing; “treadmill training” refers to using a treadmill to stimulate each group of rats after the surgical incision heals (for a total of 5 weeks), with the first week of running at a speed of 4 m / min, 10 min per day; if the balance is good, then increase to 6 m / min in the second week, 15-20 min per day; if the balance is still good in the second week, then increase to 6 m / min in the third week, 30 min per day; and maintain this intensity in the fourth to fifth weeks.

[0056] The anatomical structure of the rat paraspinal muscle is shown in Figure 1 , the surgical procedure for the double-limb model is shown in Figure 2 , the surgical procedure for the left deep paraspinal muscle removal group / control group A is shown in Figure 3 , the surgical procedure for the right deep paraspinal muscle removal group / control group B is shown in Figure 4 , and the treadmill training of the control group A / B and the left / right deep paraspinal muscle removal group rats is shown in Figure 5 .

[0057] 3. Phenotype verification and data collection (1) CT examination After treadmill training for 5 weeks, the rats in each group were anesthetized with isoflurane gas at the age of 10 weeks and 14 weeks, and then CT examination was performed.

[0058] (2) Phenotype data statistics Based on the CT examination results, the scoliosis angle, scoliosis incidence, kyphosis angle, thoracic cross diameter / thoracic diameter, and other phenotype data were calculated. At the age of 16 weeks, the rats in each group were subjected to thoracic paraspinal muscle MRI examination to confirm the effect of deep paraspinal muscle resection.

[0059] The CT examination results of the left deep paraspinal muscle resection group / control group A are shown in Figure 6 , and the phenotype data statistics results and surgical scar confirmation of the rats in each group are shown in Figure 7~8 .

[0060] The results showed that CT examination found that the rats in the left deep paraspinal muscle resection group developed right scoliosis ( Figure 5 ); the rats in the right deep paraspinal muscle resection group developed left scoliosis (see Figure 7 B for details); specifically, compared with the rats in control group A, the rats in the left deep paraspinal muscle resection group developed severe right scoliosis, kyphosis, and significant reduction in the ratio of thoracic cross diameter to thoracic diameter at the age of 10 weeks and 14 weeks ( Figure 6 ); while compared with the rats in control group B, the rats in the right deep paraspinal muscle resection group developed severe left scoliosis, kyphosis, and significant reduction in the ratio of thoracic cross diameter to thoracic diameter at the age of 10 weeks (see Figure 7 C~H for details).

[0061] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. In addition, the skilled person in the art can combine and combine the different embodiments or examples described in the present specification and the features of the different embodiments or examples without contradiction.

[0062] Although the embodiments of the present application have been shown and described above, it is understood that the above-described embodiments are exemplary and are not to be construed as limiting the present application, and that changes, modifications, substitutions and variations can be made by those skilled in the art without departing from the scope of the present application.

Claims

1. A method for constructing an animal model of scoliosis, characterized in that, The method includes the following steps: S1: Obtain the experimental animal by removing its forelimbs and tail to make its direction of gravity parallel to the spine, thus obtaining a bipedal animal. S2: The deep paravertebral muscle group on one side of the bipedal animal is removed to obtain an experimental animal with muscle mechanics imbalance. S3: The experimental animals with muscle mechanical imbalance were subjected to exercise load stimulation to obtain the scoliosis animal model; The deep paravertebral muscle group on one side includes the multifidus muscle, rotatores muscle, and semispinalis muscle; The experimental animals were tetrapod mammals.

2. The method according to claim 1, characterized in that, The resection was performed in the thoracic vertebral segment; Optionally, when the quadrupedal mammal is a rodent, the thoracic vertebral segment is at the level of the T7 to T11 vertebral bodies.

3. The method according to claim 1, characterized in that, The exercise load stimulation treatment is treadmill training; Optionally, the animal is a rodent; Optionally, the rodent is a rat.

4. The method according to claim 1, characterized in that, The experimental animals were juvenile experimental animals in the period of rapid skeletal growth.

5. The method according to claim 4, characterized in that, The childhood period refers to the period from weaning to sexual maturity.

6. An animal model of scoliosis, characterized in that, The scoliosis animal model was constructed using the method described in any one of claims 1 to 5.

7. The scoliosis animal model according to claim 6, characterized in that, The animal model of scoliosis exhibits scoliosis on the opposite side to the side of the deep paraspinal muscle group resection.

8. The scoliosis animal model according to claim 7, characterized in that, The Cobb angle of the scoliosis is greater than 10 degrees.

9. The method according to any one of claims 1 to 5, and the scoliosis animal model according to any one of claims 6 to 8, in the study of the pathogenesis of scoliosis, and in the screening or evaluation of drugs for the treatment of scoliosis.

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