Traditional Chinese medicine composition and traditional Chinese medicine preparation for treating intervertebral disc degenerative diseases of companion animals

Abstract

The invention discloses a traditional Chinese medicine composition and a traditional Chinese medicine preparation for an intervertebral disc degenerative disease of companion animals. The traditional Chinese medicine composition comprises the following traditional Chinese medicine components in parts by weight: 1-5 parts of notopterygium root, 1-5 parts of radix angelicae pubescentis, 1-5 parts of gastrodia elata, 1-5 parts of radix achyranthis bidentatae, 1-5 parts of gentiana macrophylla, 1-5 parts of caulis spatholobi, 1-10 parts of coix seed, 1-5 parts of peach kernel, 1-5 parts of safflower, 1-5 parts of radix aconiti lateralis preparata, 1-5 parts of rhizoma zingiberis and 1-5 parts of loofah sponge. The traditional Chinese medicine composition can play roles in tonifying liver and kidney, strengthening tendons and bones and improving local blood circulation in the aspect of treating the degenerative change of lumbar intervertebral disc of companion animals, and is particularly suitable for a traditional Chinese medicine iontophoresis method. The invention also provides a preparation method of the traditional Chinese medicine preparation, and application of the traditional Chinese medicine composition and the traditional Chinese medicine preparation in preparation of drugs for treating companion animal intervertebral disc degenerative diseases.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese veterinary medicine, specifically to a traditional Chinese medicine composition and preparation for treating intervertebral disc degenerative diseases in companion animals, particularly a traditional Chinese medicine composition and preparation for treating canine intervertebral disc degenerative diseases, and a method for preparing the traditional Chinese medicine preparation. Background Technology

[0002] In recent years, with the advancement of companion animal ownership concepts, the average lifespan of companion animals has gradually increased, and the prevalence of age-related diseases, chronic diseases, and degenerative diseases has also risen year by year. Among them, the incidence of intervertebral disc degeneration (IVDD) in companion animals has shown an increasing trend year by year; for example, the lifetime incidence of IVDD in dogs is about 3%.

[0003] In fact, besides companion animals such as dogs and cats, degenerative disc disease is also the most common spinal disease in humans and the leading cause of disability worldwide. Surveys show that 40% of people under 30 years old exhibit disc degeneration, while this proportion increases to approximately 80% in those over 50.

[0004] Existing research indicates that the etiology and pathogenesis of intraepithelial neoplasia (IVDD) in companion animals differ from those in humans. Human IVDD is primarily caused by prolonged poor posture or standing, leading to excessive load on the intervertebral discs, impaired nutrient supply, reduced endplate vascularization, and accelerated disc degeneration. Furthermore, human IVDD is closely related to inflammatory factors such as IL-1β, TNF-α, and PGE2, which promote the secretion of matrix metalloproteinases (MMPs) through activation of the NF-κB pathway, accelerating extracellular matrix degradation. Oxidative stress and apoptosis are also key mechanisms in the pathogenesis of human IVDD. In contrast, the prone gait of terrestrial non-primate quadrupedal mammals such as dogs and cats results in significantly different intervertebral disc stress patterns compared to humans, leading to distinct pathogenesis mechanisms. For instance, small dogs like Poodles and Bulldogs, with their smaller epidural space and congenital dysplasia of the intervertebral disc annulus fibrosus, are more susceptible to mechanical compression leading to nucleus pulposus protrusion, thus making them prone to canine IVDD. In addition, aging-related decreases in nucleus pulposus water content and annulus fibrosus elasticity are major contributing factors to IVDD in companion animals; for example, the incidence of IVDD is significantly higher in dogs over 6 years of age. Furthermore, canine IVDD is more characterized by anatomical abnormalities and some inflammatory responses, while human IVDD is primarily characterized by chronic inflammation and metabolic disorders, with genetic and environmental interactions playing a significant role.

[0005] Besides the pathogenesis, the classification of IVDD in companion animals differs from that in humans. For example, canine IVDD uses the Hansen classification, which mainly divides it into four types: Hansen type I is acute disc herniation, often seen in young dogs after trauma or strenuous activity; Hansen type II is often seen in chronic disc degeneration, more common in older dogs; Hansen type III is discospondyloarthritis; and Hansen type IV represents mild spinal cord compression due to various causes. Human IVDD, however, has a more complex clinical classification. Based on the stage of disease progression and the degree of spinal cord compression, it is divided into pre-disc herniation / disc bulging, disc herniation, and disc extrusion. Based on the location of the herniated disc, it can be further divided into lateral and central herniation types. Lateral herniation can be further divided into root-shoulder, root-axillary, pre-root, and far lateral types, while central herniation can be divided into off-central and central types. In summary, canine IVDD classification focuses on clinical manifestations and the degree of acuteness, while human IVDD classification is more complex.

[0006] Given the significant differences in pathogenesis and disease subtypes, current medications for human IVDD have limited efficacy in treating IVDD in companion animals. Furthermore, there is considerable controversy and disagreement regarding the choice of treatment methods for canine IVDD. Opinions differ on the necessity of surgery. Surgery is effective for acute disc herniation, but has a strict treatment window, typically requiring surgery within 24 to 48 hours of onset. Its effectiveness decreases significantly in dogs where deep pain has subsided, and assessing the duration of deep pain disappearance is difficult in affected animals. For dogs with only mild neurological symptoms such as lower back pain or hyperesthesia, early non-surgical treatment often yields good results. Traditional conservative treatments include restricting movement, anti-inflammatory drugs, and analgesia. However, some studies have shown that when companion animals have disc herniation symptoms, the use of anti-inflammatory drugs without immobilization may lead to increased activity due to reduced pain, potentially exacerbating pressure from adjacent vertebrae on the herniated area. This could cause more disc contents to be compressed into the spinal canal, worsening clinical symptoms.

[0007] Traditional Chinese medicine (TCM) iontophoresis is a treatment method that utilizes a direct current electric field or a low-frequency pulsed electric field to ionize molecules in a TCM liquid, allowing them to enter the body through the skin or mucous membranes. Practice has shown that, as a conservative treatment, this therapy allows medication to directly reach the affected area, effectively combining drug therapy, acupoint therapy, and electrical physical therapy to quickly relieve pain symptoms. However, currently, there is a lack of TCM compositions and preparations suitable for TCM iontophoresis with proven efficacy against IVDD in companion animals. Summary of the Invention

[0008] This invention provides a traditional Chinese medicine composition and preparation for treating intervertebral disc degenerative diseases in companion animals. This composition and preparation, combined with iontophoresis, can effectively and safely treat intervertebral disc degenerative diseases in companion animals such as dogs and cats. This invention also provides a method for preparing the aforementioned traditional Chinese medicine preparation, as well as applications of the composition and preparation.

[0009] To achieve the above objectives, a first aspect of the present invention is to provide a traditional Chinese medicine composition for intervertebral disc degenerative diseases in companion animals, comprising the following traditional Chinese medicine components in parts by weight:

[0010] Notopterygium root 1-5 parts, Angelica pubescens root 1-5 parts, Gastrodia elata root 1-5 parts, Achyranthes bidentata root 1-5 parts, Gentiana macrophylla root 1-5 parts, Spatholobus suberectus root 1-5 parts, Coix seed 1-10 parts, Peach kernel 1-5 parts, Carthamus tinctorius root 1-5 parts, Prepared aconite root 1-5 parts, Dried ginger 1-5 parts, Luffa cylindrica root 1-5 parts.

[0011] The main active ingredients of the traditional Chinese medicine composition provided by this invention include Notopterygium root, Angelica pubescens root, and Achyranthes root, whose main functions are to dispel wind, cold, dampness, and pain. They are suitable for symptoms such as rheumatic pain. According to the Chinese Pharmacopoeia, the above-mentioned medicinal materials all have anti-inflammatory and analgesic effects, and can effectively relieve pain and swelling caused by inflammation. Among them, Notopterygium root relieves exterior syndromes, dispels cold, eliminates wind and dampness, and relieves pain, mainly used for wind-cold-dampness arthralgia; Angelica pubescens dispels wind and dampness, relieves pain, and is good for treating soreness and pain in the waist and knees; Achyranthes root invigorates blood circulation, regulates menstruation, tonifies the liver and kidneys, and strengthens tendons and bones, and is often used for soreness and pain in the waist and knees, and weakness and paralysis of the lower limbs. Gastrodia elata calms the liver, extinguishes wind, and relieves pain, and its components such as gastrodin have a protective effect on the nervous system. Gentiana macrophylla root, Spatholobus suberectus root, and Coix seed respectively play the roles of dispelling wind and dampness, relaxing tendons and collaterals, clearing damp heat, invigorating blood circulation, replenishing blood, regulating menstruation, and relieving pain. At the same time, Coix seed can also strengthen the spleen and eliminate dampness. The formula combines peach kernel, safflower, prepared aconite root, dried ginger, and loofah sponge. Peach kernel and safflower work together to invigorate blood and remove blood stasis; prepared aconite root restores yang and reverses collapse, tonifies fire and assists yang; dried ginger warms the middle and dispels cold; and loofah sponge unblocks meridians and activates collaterals. While the toxicity of prepared aconite root is reduced after processing, it retains its strong warming properties and can enhance the body's metabolic function. Dried ginger stimulates the secretion of digestive enzymes, increases gastrointestinal motility, and also has a certain anti-inflammatory effect. Through the rational combination of principal, assistant, and adjuvant herbs, and utilizing the unique characteristics and synergistic effects of each herb, the formula provided by this invention can tonify the liver and kidneys, strengthen tendons and bones, and improve local blood circulation in the treatment of degenerative lumbar disc disease in dogs.

[0012] The aforementioned traditional Chinese medicine composition is particularly suitable for iontophoresis, demonstrating significant therapeutic effects on intravertebral disc degeneration (IVDD) in companion animals, especially canine and feline IVDD. Furthermore, by optimizing the proportions of the herbal composition, the advantages of multi-component, multi-target, and multi-pathway interactions of traditional Chinese medicine can be further leveraged to promote liver and kidney tonification, strengthen tendons and bones, improve local blood circulation, and control inflammation. Therefore, it is of great significance for the clinical treatment of canine lumbar intervertebral disc degeneration. In specific implementation, the herbal composition includes the following components by weight: 3-5 parts of Notopterygium root, 3-5 parts of Angelica pubescens root, 3-5 parts of Gastrodia elata root, 3-5 parts of Achyranthes bidentata root, 2-4 parts of Gentiana macrophylla root, 2-4 parts of Spatholobus suberectus root, 4-7 parts of Coix seed, 1-3 parts of Prunus persica seed, 1-3 parts of Carthamus tinctorius flower, 1-3 parts of processed Aconitum carmichaelii root, 1-3 parts of dried ginger, and 1-3 parts of Luffa cylindrica root. In practice, the drug composition can be adjusted based on real-time feedback from the affected animal to achieve the best therapeutic effect. For example, in one feasible embodiment, the following traditional Chinese medicine formula can be used: 4 parts of Qianghuo, 4 parts of Duhuo, 4 parts of Tianma, 4 parts of Niuxi, 3 parts of Qinjiu, 3 parts of Jixueteng, 5 parts of Yiyiren, 2 parts of Taoren, 2 parts of Honghua, 2 parts of Zhifuzi, 2 parts of Ganjiang, and 2 parts of Sigualu.

[0013] In addition to adjusting the dosage of the Chinese herbal medicine composition according to the symptoms, other Chinese herbal medicine components can be added according to the actual condition of the sick animal. For example, 3-6 parts of Eucommia ulmoides and 3-6 parts of Loranthus parasiticus can be added to the above-mentioned Chinese herbal medicine composition to further tonify the liver and kidneys and strengthen muscles and bones; or 3-6 parts of Corydalis yanhusuo, 1-3 parts of Tripterygium wilfordii, and 1-3 parts of Buthus martensii can be added to the above-mentioned Chinese herbal medicine composition to enhance anti-inflammatory and analgesic effects. Of course, Tripterygium wilfordii needs to be decocted for a long time to reduce toxicity; or 6-10 parts of Astragalus membranaceus and 3-6 parts of Angelica sinensis can be added to the above-mentioned Chinese herbal medicine composition to enhance the effects of tonifying qi and blood, and strengthening the body's resistance.

[0014] A second aspect of the present invention is to provide a traditional Chinese medicine preparation for intervertebral disc degeneration in companion animals, the preparation comprising a powder of the above-mentioned traditional Chinese medicine composition or an extract of the traditional Chinese medicine composition.

[0015] The traditional Chinese medicine preparation provided by this invention is for external use. Its dosage form can be any common external dosage form, such as liquid, semi-solid, gaseous, or solid dosage forms. Liquid preparations, such as decoctions, tinctures, and extracts, can increase drug dispersibility and reduce the irritation of certain drugs. Semi-solid dosage forms, such as pastes, ointments, oils, and gels, have a uniform and delicate appearance and possess adhesiveness and chemical stability. Gaseous dosage forms, such as sprays and aerosols, have stabilizing, rapid-acting, and targeted effects. Solid dosage forms, such as powders, allow for dosage adjustment based on symptom severity, and solid dosage forms are convenient to transport, carry, and use, have simple preparation methods, and high utilization rates of medicinal materials.

[0016] As mentioned above, the traditional Chinese medicine composition provided by this invention is particularly suitable for iontophoresis of traditional Chinese medicine. Considering that liquid preparations have good conductivity when administered via iontophoresis of traditional Chinese medicine, which is conducive to the effective absorption of the active ingredients in the drug and the rapid exertion of therapeutic effects, they can serve as an effective medium for iontophoresis of traditional Chinese medicine. Therefore, in preferred embodiments, the traditional Chinese medicine preparation provided by this invention is a liquid preparation, such as a decoction with an aqueous matrix, and especially a water decoction.

[0017] Aqueous decoctions are made by boiling Chinese medicinal herbs in water, removing the residue, and extracting the juice. They are characterized by rapid absorption, strong effects, and simple preparation. Furthermore, compared to other dosage forms such as tinctures and extracts, aqueous decoctions exhibit significant advantages when used with iontophoresis. First, the active ingredients in aqueous decoctions are mostly in ionic form, meeting the requirements of iontophoresis technology, and can effectively allow charged drug ions to enter the body using a direct current electric field. Second, compared to alcohol-containing preparations, aqueous decoctions are safer, reducing the risk of skin irritation and allergies. Third, aqueous decoctions can be prepared immediately, and the drug composition can be flexibly adjusted according to the actual condition of the sick animal to meet personalized treatment needs and achieve the best therapeutic effect.

[0018] The third aspect of this invention provides a method for preparing a traditional Chinese medicine preparation, which uses the traditional Chinese medicine composition described in the first aspect as raw material.

[0019] This traditional Chinese medicine preparation may contain extracts or powders of a traditional Chinese medicine composition. The preparation method varies depending on the dosage form of the preparation. Taking a decoction as an example, the traditional Chinese medicine composition is first ground or chopped into coarse powder, then soaked in water to obtain a pre-treated solution, and finally decocted to obtain a decoction (or soup).

[0020] In the above process, each Chinese herbal medicine component can first be ground or chopped into coarse powder (see the definition of coarse powder in the Chinese Pharmacopoeia for details). Then, according to the required mass proportions, take the coarse powder of each herb and soak it separately in cold water. The water used should be clean drinking water, such as purified water or distilled water, to avoid introducing impurities. The amount of soaking water can be 5 to 10 times the mass of each coarse powder. If pre-decoction is required or the pre-decoction time is long, the amount of soaking water can be increased accordingly. The soaking time can be more than 20 minutes, preferably 30 to 60 minutes, to ensure that the effective ingredients are fully dissolved. Among them, the pre-treatment liquid obtained after soaking Gastrodia elata and Coix lacryma-jobi should be filtered and wrapped in gauze for decoction to avoid turbidity. It should be noted that although it is called pre-treatment "liquid", it actually includes water, the effective ingredients dissolved in the herbs during the soaking process, and the herbs soaked in water.

[0021] The pre-treated solutions of *Spatholobus suberectus* and processed aconite root need to be decocted separately to obtain a pre-decoction. *Spatholobus suberectus* is hard and requires decocting for 20-30 minutes; processed aconite root contains aconitine and requires decocting for 60-120 minutes to reduce toxicity. The pre-treated solutions of the remaining herbs are mixed and brought to a boil over high heat. Then, the *Gastrodia elata*, *Coix lacryma-jobi* (wrapped in a decoction packet), and the pre-decoction solutions of *Spatholobus suberectus* and processed aconite root are added, followed by simmering over low heat for 20-30 minutes. After the first decoction, the dregs are filtered through gauze or a strainer, and the first decoction liquid is collected. The dregs are then mixed and used for the second decoction. The second decoction is again brought to a boil over high heat, followed by simmering over low heat for 15-20 minutes. The first and second decoction liquids are then combined to prepare the decoction. Ceramic, earthenware, or stainless steel pots are suitable for decoction; iron or aluminum pots should not be used. Of course, a third decoction can be performed after the second decoction. In addition, during the above decoction process, the mass ratio of medicinal materials to water can be controlled at 1:(5-10) before simmering over low heat.

[0022] The fourth aspect of the present invention provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a medicament for treating intervertebral disc degeneration in companion animals; the fifth aspect provides the use of the above-mentioned traditional Chinese medicine preparation in the preparation of a medicament for treating intervertebral disc degeneration in companion animals.

[0023] This traditional Chinese medicine composition and preparation are particularly suitable for iontophoresis. In practice, a direct current electric field is used to ionize the aforementioned traditional Chinese medicine preparation, especially the decoction, and introduce it into the body through the skin or mucous membranes for the treatment of canine intervertebral disc degeneration. Specifically, the decoction of the aforementioned traditional Chinese medicine composition can be used as an external application, covered with gauze or similar material onto an electrode plate. The affected area of ​​the animal is shaved and prepared, and the electrode plate is applied to the Jiaji acupoints and Ashi acupoints on both sides of the affected area. A moderate heat parameter is selected, ranging from 37-43℃, and a moderate intensity parameter is selected, ranging from 5-10mA. Each treatment lasts 15 minutes, with a frequency of once every 1-3 days, and 7 treatments constitute one course of treatment. The number of treatment courses can be increased or decreased according to the therapeutic effect.

[0024] Evaluation results showed that electroacupuncture and traditional Chinese medicine iontophoresis could alleviate intervertebral disc narrowing, reduce endplate sclerosis and osteophyte formation, alleviate decreased nucleus pulposus hydration, and inhibit intervertebral disc degeneration and cell apoptosis. Among these, the traditional Chinese medicine iontophoresis therapy using a decoction was convenient, flexible in regimen adjustment, and showed high compliance in affected animals. It also had a rapid onset of action, a short treatment cycle, and effectively tonified the liver and kidneys, strengthened muscles and bones, improved local blood circulation, and controlled inflammation.

[0025] The traditional Chinese medicine composition and preparation provided by this invention for treating intervertebral disc degeneration in companion animals, through the combination of various Chinese medicine components, can tonify the liver and kidneys, strengthen tendons and bones, and improve local blood circulation in the treatment of lumbar intervertebral disc degeneration in companion animals, and is particularly suitable for the Chinese medicine iontophoresis method. The preparation method of the Chinese medicine preparation provided by this invention is simple and easy to apply and promote in practice. Because the above-mentioned Chinese medicine composition and preparation have the effects described above, they can be applied to the treatment of intervertebral disc degeneration in companion animals, such as canine lumbar intervertebral disc degeneration, and have a very broad application prospect. Attached Figure Description

[0026] Figure 1 These are photographs showing the depth of needle insertion and the selection of puncture sites when using the CT-guided percutaneous annulus fibrosus puncture and nucleus pulposus aspiration method to establish a rabbit lumbar intervertebral disc degeneration model in the experimental examples of this invention.

[0027] Figure 2 This is a diagram showing the results of the evaluation model established according to the MRI Pfirmann grading system in the experimental examples of this invention.

[0028] Figure 3 These are photographs of the electroacupuncture (EA) treatment group in the experimental examples of this invention;

[0029] Figure 4 These are photographs of the treatment status of the traditional Chinese medicine iontophoresis group (ICM) in the experimental examples of this invention;

[0030] Figure 5 This is a schematic diagram illustrating the MRI index calculation method in the experimental examples of this invention;

[0031] Figure 6 The Weishuapt grading scores for each group in the experimental examples of this invention are shown below.

[0032] Figure 7 These are the Pfirmann grading scores for each group in the experimental examples of this invention.

[0033] Figure 8 HE staining images (×100) of the model control group (MC), local block group (PC), electroacupuncture treatment group (EA), and traditional Chinese medicine iontophoresis group (ICM) in the experimental examples of this invention. Detailed Implementation

[0034] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, 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 not all embodiments of the present invention. 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.

[0035] Example 1

[0036] This embodiment provides a traditional Chinese medicine composition, the formula of which is: 4 parts of Qianghuo, 4 parts of Duhuo, 4 parts of Tianma, 4 parts of Niuxi, 3 parts of Qinjiu, 3 parts of Jixueteng, 5 parts of Yiyiren, 2 parts of Taoren, 2 parts of Honghua, 2 parts of Zhifuzi, 2 parts of Ganjiang, and 2 parts of Sigualu.

[0037] Example 2

[0038] This embodiment provides a traditional Chinese medicine preparation, the preparation process of which specifically includes the following steps:

[0039] Notopterygium root, Angelica pubescens root, Gastrodia elata root, Achyranthes bidentata root, Gentiana macrophylla root, Spatholobus suberectus root, Coix seed, Prunus persica seed, Carthamus tinctorius flower, processed Aconitum carmichaelii root, dried ginger, and Luffa cylindrica root are ground into coarse powder. Then, according to the proportions in Example 1, the coarse powder of each Chinese herbal component is taken separately and soaked in clean drinking water to obtain the corresponding pretreatment solution of the Chinese herbal component. The amount of water used for soaking can be 5 to 10 times the mass of each coarse powder, and the soaking time can be 30 to 60 minutes. The pretreatment solution obtained after soaking Gastrodia elata root and Coix seed is first filtered through the herbs and wrapped in gauze.

[0040] First, decoct the chicken blood vine for 25 minutes and the prepared aconite root for 100 minutes. Mix the pre-treated liquids of the remaining herbs and bring to a boil over high heat. Then add the gastrodia elata, coix seed (wrapped in a decoction packet), and the pre-decoctions of chicken blood vine and prepared aconite root. Reduce the heat to low and simmer for 30 minutes, monitoring the liquid level to prevent it from drying out or overflowing. After the first decoction, filter the dregs with gauze or a strainer and collect the first decoction. Mix the dregs together and decoct a second time. Again, bring to a boil over high heat, then simmer over low heat for 18 minutes. Combine the first and second decoctions to obtain the final decoction.

[0041] Experimental Example

[0042] 1. New Zealand white rabbits were selected as the model animal, and a rabbit lumbar intervertebral disc degeneration model was established by CT-guided percutaneous annulus fibrosus puncture and nucleus pulposus aspiration.

[0043] New Zealand white rabbits weighing 2.5-3.0 kg were selected as the model animal, regardless of sex, free from acute and chronic infectious diseases and immune system diseases, with good limb movement, and no spinal deformities or intervertebral disc diseases. The model was established after 3 days of single-cage rearing. Preoperatively, rabbits were fasted for 6 hours and dehydrated for 4 hours. Sedation was initiated with a 20 μg / kg intramuscular injection of dexmedetomidine. Anesthesia was established via the marginal ear vein 10-15 minutes after administration, followed by propofol 0.1-0.5 mg / kg / h CRI maintenance anesthesia. After confirming the anesthesia status, the rabbits were placed in the left lateral decubitus position, and the surgical field was prepared and disinfected. A 21G needle was used, and under CT guidance, the L4-L5 intervertebral disc was located as the puncture point. The needle was inserted vertically to puncture the skin and subcutaneous tissue, deflecting towards the medial aspect of the head to penetrate the deep muscle. CT scans were performed during the procedure to confirm the puncture segment and location. Continue advancing the needle. Significant resistance indicates that the spine has been reached. Under CT guidance, adjust the needle tip to the level of the intervertebral space and puncture into the annulus fibrosus. The annulus fibrosus is relatively tough, and the feel is similar to a bone marrow aspiration. A feeling of emptiness after penetrating the annulus fibrosus indicates that the needle tip has entered the nucleus pulposus. Stop the puncture and perform a CT scan to confirm a puncture depth of approximately 5mm. Figure 1 As shown, the left image is a photograph of the depth of needle insertion, and the right image is a photograph of the puncture site selection. The needle was left in place for 30 seconds, and a 5mL syringe was used to draw 3mL of negative pressure, which was maintained for 15 seconds before the needle was withdrawn for nucleus pulposus aspiration. Postoperatively, the animal's mental state, limb mobility, appetite, bowel movements, and other general conditions were observed.

[0044] 2. After modeling, the Pfirmann grading system was used to score the degree of intervertebral disc degeneration and evaluate the modeling effect:

[0045] Four weeks after modeling, the rabbit lumbar spine underwent CT and MRI scans. For CT scans, the tube voltage was 140 kV, tube current 160 mA, matrix 512×512, slice thickness 0.75 mm, reconstruction interval 0.375 mm, collimation 16×0.75, pitch 0.6042, and rotation time 0.75 s. Bone window (WL: 500 HU, WW: 1600) images were constructed using the bone filtering algorithm (EB), and soft tissue window (WL: 40 HU, WW: 350) images were constructed using the soft tissue filtering algorithm (SB). MRI scan sequences included T2-weighted (T2WI) sagittal images, T1-weighted (T1WI) sagittal images, T2WI axial images, DWI sagittal images, and T2 mapping sagittal images. Commonly used scanning parameters are T2WI sagittal plane (TR: 2500.0 ms, TE: 103.0 ms), T1WI sagittal plane (TR: 395.0 ms, TE: 11.0 ms), and T2W axial plane (TR: 3000.0 ms, TE: 91.0 ms). The intervertebral disc degeneration in the T2WI sagittal plane was graded according to the MRI Pfirmann grading system to evaluate the model establishment effect. The specific classifications are as follows: Grade I: Uniform and bright white nucleus pulposus structure, clear boundary between the nucleus pulposus and the annulus fibrosus, high signal intensity of the nucleus pulposus, and normal disc height; Grade II: Heterogeneous nucleus pulposus structure, possibly with horizontal bands, clear boundary between the nucleus pulposus and the annulus fibrosus, high signal intensity of the nucleus pulposus, and normal disc height; Grade III: Heterogeneous and gray nucleus pulposus structure, unclear boundary between the nucleus pulposus and the annulus fibrosus, heavy signal intensity of the nucleus pulposus, and slightly reduced disc height; Grade IV: Heterogeneous and gray to black nucleus pulposus structure, loss of boundary between the nucleus pulposus and the annulus fibrosus, medium to low signal intensity of the nucleus pulposus, and moderately reduced disc height; Grade V: Heterogeneous and black nucleus pulposus structure, loss of boundary between the nucleus pulposus and the annulus fibrosus, low signal intensity of the nucleus pulposus, and severely reduced disc height.

[0046] like Figure 2 As shown, the upper left and lower left images in the figure are sagittal MRI T2WI images of the lumbar spine before and after modeling, respectively. The upper right image is a cross-sectional view of the intervertebral disc at the L4-L5 modeling site, and the lower right image is a cross-sectional view of the intervertebral disc at the L5-L6 control site. Figure 2 As can be seen, the nucleus pulposus of the intervertebral disc showed normal high signal on T2WI before modeling, and the T2WI signal of the nucleus pulposus decreased after modeling, while the Pfirmann score increased, indicating successful modeling.

[0047] 3. After modeling and evaluation, treat the experimental animals:

[0048] The experimental animals were randomly divided into four groups: model control group (MC), partial closure of the treatment group (PC), electroacupuncture treatment group (EA), and iontophoresis of Chinese medicine group (ICM).

[0049] The model control group (MC) was housed in single cages with restricted movement after modeling and received no other treatment. The local injection group (PC) received dexamethasone 0.5 mg / kg, lidocaine 2 mg / kg, and ampicillin sodium 50 mg / kg dissolved in 1 mL of water for injection; intramuscular injection was administered bilaterally from L3 to L6 for 3 consecutive days. The electroacupuncture group (EA) received acupuncture at the following acupoints: Shenshu (BL23), Qihaiyu (BL60), Dachangshu (BL25), Guanyuanshu (BL24), Yaobaihui (GV20), and Weizhong (BL40). The electroacupuncture connection was ipsilateral Shenshu-Guanyuanshu and Yaobaihui-Weizhong, with parameters of 0.5-2.5 mV, alternating between 2 Hz 5s and 15 Hz 3s, 20 minutes per session, 3 days per session, for 7 treatments. Photos taken at different angles and magnifications during treatment are shown below. Figure 3 As shown. The traditional Chinese medicine iontophoresis group (ICM) used the decoction from Example 2; the instrument parameters were: heat 2, intensity 2; 15 minutes per session, 3 days / session, 7 treatments in total. Multiple photos during the treatment process are shown below. Figure 4 As shown.

[0050] 4. Assess the differences in radiographic findings among the experimental groups:

[0051] Following modeling and treatment, regular CT and MRI scans were performed to quantitatively assess disc height change (DHI) and disc height index percentage (DHI%). Facet joint degeneration was assessed using the Weishuapt grading system, which classifies it into four grades: Grade 0: normal facet joint space; Grade 1: narrowing of the facet joint space and / or mild osteophyte formation and / or mild facet joint hypertrophy; Grade 2: narrowing of the facet joint space and / or moderate osteophyte formation and / or moderate facet joint hypertrophy and / or mild subarticular bone erosion; Grade 3: narrowing of the facet joint space and / or severe osteophyte formation and / or severe facet joint hypertrophy and / or severe subarticular bone erosion and / or subchondral cysts. The Pfirmann grading system was used to grade T2WI sagittal disc degeneration, with the grading criteria as described above. MRI was used to assess the degree of disc degeneration, spinal cord compression, paravertebral muscle signal changes, and spinal cord signal alterations. MRI indices were used to quantitatively evaluate the degree of lumbar disc degeneration. The MRI index is the product of the area of ​​the nucleus pulposus and its corresponding signal intensity on T2WI sagittal plane, such as... Figure 5 As shown, Figure A is the T2WI sagittal signal map of the rabbit intervertebral disc, and Figure B is the area, signal intensity, and MRI index of the nucleus pulposus measured after delineating the region of interest (ROI). Figure 6 and Figure 7 The results are Weishuapt and Pfirmann grading scores for each group before and after modeling.

[0052] Four weeks after modeling, there were no statistically significant differences among the groups in DHI%, Weishuapt grade, Pfirmann grade, MRI index, and T2 mapping of the anterior annulus fibrosus (AAF) and posterior annulus fibrosus (PAF) T2 values ​​(P>0.05).

[0053] Eight weeks after modeling, compared with the model control group (MC) and the local block group (PC), the ADC values ​​of the electroacupuncture treatment group (EA) and the traditional Chinese medicine iontophoresis group (ICM) increased, and the T2 value of the T2 mapping nucleus pulposus (NP) increased, with statistically significant differences (P<0.05).

[0054] Twelve weeks after modeling, compared with the model control group (MC) and the local block group (PC), the DHI% of the electroacupuncture treatment group (EA) and the traditional Chinese medicine iontophoresis group (ICM) increased, the Weishuapt grade decreased, the Pfirmann grade decreased, the MRI index increased, the ADC value increased, and the T2 value of T2 mapping NP increased, with statistically significant differences (P<0.05).

[0055] After 12 weeks of modeling, there were no statistically significant differences in DHI%, Weishaopt grade, Pfirmann grade, MRI index, ADC value, T2 mapping AAF, and PAF T2 value between the model control group (MC) and the local injection group (PC) (P>0.05). The T2 mapping NP T2 value was significantly higher in the local injection group than in the model control group (MC) (P<0.05). Similarly, there were no statistically significant differences in DHI%, Weishaopt grade, Pfirmann grade, MRI index, T2 mapping AAF, and PAF T2 value between the electroacupuncture group (EA) and the traditional Chinese medicine iontophoresis group (ICM) (P>0.05). The ADC value and T2 mapping NP T2 value were significantly higher in the traditional Chinese medicine iontophoresis group than in the electroacupuncture group (EA) (P<0.05).

[0056] After 12 weeks of modeling, compared with before treatment, the DHI% and Weishuapt grade, Pfirmann grade, MRI index, ADC value, and T2 value of T2 mapping NP of the model control group (MC) and the local block group (PC) increased, and the differences were statistically significant (P<0.05).

[0057] 5. After treatment, the animals were euthanized, and their tissues were fixed and stained with hematoxylin and eosin (HE) to evaluate the pathological differences between the treatment groups and to clarify the therapeutic effect of the traditional Chinese medicine iontophoresis method.

[0058] Twelve weeks after modeling, the experimental animals were sacrificed, and tissues from the L4-L5 intervertebral disc annulus fibrosus and nucleus pulposus of each experimental group were subjected to HE staining. The results are as follows: Figure 8 As shown in the image, normal intervertebral disc tissue appears as a slightly bluish, transparent, gelatinous substance with the annulus fibrosus encompassing the nucleus pulposus. After modeling, the intervertebral disc tissue gradually darkens and becomes cloudy, its volume gradually shrinks, the boundary between the annulus fibrosus and nucleus pulposus becomes unclear, and the water content of the nucleus pulposus is significantly reduced. HE staining revealed that in the model control group (MC), the boundary between the annulus fibrosus and nucleus pulposus structure was interrupted, the number of nucleus pulposus cells in the nucleus pulposus tissue decreased, vacuoles disappeared, the nucleus pulposus matrix was moderately to severely compressed, and the annulus fibrosus was broken or twisted. In contrast, the local injection group (PC), electroacupuncture group (EA), and traditional Chinese medicine iontophoresis group (ICM) all showed varying degrees of relief from the above-mentioned lesions.

[0059] The Masuda Intervertebral Disc Degeneration Pathological Scoring Criterion was used for quantitative assessment, specifically evaluating the following four aspects: 1 point for normal annulus fibrosus morphology, 2 points for fiber breakage or tortuosity <30%, and 3 points for fiber breakage or tortuosity ≥30%. 1 point for a normal boundary between the annulus fibrosus and nucleus pulposus, 2 points for mild interruption, and 3 points for moderate to severe interruption. 1 point for normal nucleus pulposus cell count with abundant vacuoles, 2 points for mild cell reduction and smaller vacuoles, and 3 points for moderate to severe cell reduction and disappearance of vacuoles. 1 point for a gel-like nucleus pulposus matrix, 2 points for mild compression, and 3 points for moderate to severe compression. The average scores for each group were as follows: Model Control Group (MC) 11.2 points, Local Injection Group (PC) 7.8 points, Electroacupuncture Group (EA) 5.6 points, and Traditional Chinese Medicine Iontophoresis Group (ICM) 5.4 points. Clearly, the Traditional Chinese Medicine Iontophoresis Group (ICM) had the lowest score and the best effect.

[0060] Post-treatment evaluation results showed that, compared with the model control group (MC) and the local injection group (PC), electroacupuncture and traditional Chinese medicine iontophoresis could alleviate the degree of intervertebral space narrowing, reduce the degree of endplate osteosclerosis and osteophyte formation, alleviate the degree of decreased nucleus pulposus hydration, and inhibit intervertebral disc degeneration and cell apoptosis. Among them, the traditional Chinese medicine iontophoresis therapy using the decoction in Example 2 is convenient, flexible in treatment plan adjustment, has strong animal compliance, rapid onset of action, and short treatment cycle, and can effectively achieve the effects of tonifying the liver and kidneys, strengthening tendons and bones, improving local blood circulation, and controlling inflammation.

[0061] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A traditional Chinese medicine composition for treating intervertebral disc degenerative diseases in companion animals, characterized in that, It includes the following Chinese herbal medicine components in parts by weight: Notopterygium root 1-5 parts, Angelica pubescens root 1-5 parts, Gastrodia elata root 1-5 parts, Achyranthes bidentata root 1-5 parts, Gentiana macrophylla root 1-5 parts, Spatholobus suberectus root 1-5 parts, Coix seed 1-10 parts, Peach kernel 1-5 parts, Carthamus tinctorius root 1-5 parts, Prepared aconite root 1-5 parts, Dried ginger 1-5 parts, Luffa cylindrica root 1-5 parts.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, It includes the following components in parts by weight: Notopterygium root 3-5 parts, Angelica pubescens root 3-5 parts, Gastrodia elata root 3-5 parts, Achyranthes bidentata root 3-5 parts, Gentiana macrophylla root 2-4 parts, Spatholobus suberectus root 2-4 parts, Coix seed 4-7 parts, Peach kernel 1-3 parts, Carthamus tinctorius flower 1-3 parts, Prepared aconite root 1-3 parts, Dried ginger 1-3 parts, Luffa cylindrica root 1-3 parts.

3. A traditional Chinese medicine preparation for treating intervertebral disc degenerative diseases in companion animals, characterized in that, The traditional Chinese medicine preparation includes an extract of the traditional Chinese medicine composition according to claim 1 or 2, and is a liquid preparation.

4. The traditional Chinese medicine preparation according to claim 3, characterized in that, The liquid dosage form is a decoction, and the base of the decoction is an aqueous base.

5. A method for preparing a traditional Chinese medicine preparation according to claim 3 or 4, characterized in that, This includes the step of preparing the traditional Chinese medicine components into the target dosage form.

6. The preparation method according to claim 5, characterized in that, Includes the following steps: The coarse powders of each Chinese herbal medicine component were soaked in water for at least 20 minutes to obtain the pretreatment solutions of each Chinese herbal medicine component. The pretreatment solutions of Gastrodia elata and Coix lacryma-jobi were filtered and the medicinal materials were wrapped in gauze. Chicken blood vine pretreatment solution and processed aconite pretreatment solution were decocted separately to obtain chicken blood vine decoction and processed aconite decoction respectively; Mix the pretreatment solutions except for the chicken blood vine pretreatment solution and the prepared aconite pretreatment solution, bring to a boil over high heat, then add the gastrodia elata decoction packet and coix seed decoction packet, as well as the chicken blood vine pre-decoction solution and the prepared aconite pre-decoction solution, then simmer over low heat, and collect the first decoction liquid and dregs. The dregs should be boiled with water at least once, and each time the decoction should be brought to a boil over high heat and then simmered over low heat. Combine the decoctions from each decoction to obtain the decoction.

7. The preparation method according to claim 6, characterized in that, The pretreatment solution is obtained by adding water in a volume of 5-10 ml based on the total mass of the coarse powder; In obtaining the first decoction, simmer over low heat for 20-30 minutes; in obtaining subsequent decoctions, simmer over low heat for 15-20 minutes.

8. The preparation method according to claim 6, characterized in that, Simmer the chicken blood vine pretreatment solution for 20-30 minutes, and simmer the aconite pretreatment solution for 60-120 minutes.

9. The use of the traditional Chinese medicine composition according to claim 1 or 2 in the preparation of a medicament for treating intervertebral disc degeneration in companion animals.

10. The use of any one of the traditional Chinese medicine preparations according to claims 3-5 in the preparation of a medicament for treating intervertebral disc degeneration in companion animals.

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