Premix for improving joint health of pets and preparation method thereof

Abstract

The invention belongs to the field of pet food processing, and relates to a premix for improving joint health of pets and a preparation method thereof. The premix is added with the following natural components: rhizoma polygonati, rhizoma drynariae, eucommia ulmoides, ligusticum wallichii, liquorice, turmeric, poria cocos and Chinese yam. The whole formula has the effects of tonifying liver and kidney to consolidate the root, promoting blood circulation to remove meridian obstruction to treat symptoms, tonifying spleen to remove dampness to assist transportation and transformation, tonifying spleen to prevent dampness stagnation, promoting diuresis to prevent positive qi injury, treating both symptoms and root causes, combining dredging and tonifying, and directly reaching the effects on joints. The premix can tonify cartilage, strengthen ligament, relieve pain, supplement synovial fluid, effectively improve the arthritis health of pets and improve the life quality of pets.

Description

Technical Field

[0001] This invention belongs to the field of pet food processing and relates to a premixed feed for improving the joint health of pets and its preparation method. Background Technology

[0002] As people pay increasing attention to health and quality of life, pet health is also receiving more attention. As pets age, their organs and physiological functions undergo a series of degenerations, making them susceptible to various diseases, among which skeletal diseases are common in pets.

[0003] The main characteristics of osteoarthritis in dogs and cats include degeneration of articular cartilage, collagen degradation, and cartilage sclerosis. Osteoarthritis can cause symptoms such as hind limb weakness and lameness in dogs and cats. The incidence of osteoarthritis varies among different age groups and is also related to the dog's or cat's weight and genetic factors.

[0004] Osteoarthritis usually results from the degeneration of articular cartilage, especially when there are osteophytes, bone remodeling, and significant deformation of joint tissues. This can easily lead to excessive wear and tear on the articular cartilage. In particular, the cell density around the joints of pets is relatively low, and the long-term wear and tear between these cells and high-density cells is a major pathogenic mechanism of osteoarthritis. Since the bones and joints of dogs and cats are mainly composed of type II collagen and proteoglycans, the growth and development of these structures and nutrients are relatively slow. This leads to the asynchronous development of cartilage structures in dogs and cats, resulting in articular cartilage degeneration.

[0005] With advancements in technology and in-depth research, people have begun to explore the application potential of traditional Chinese medicine in the field of pet health. Traditional Chinese medicine is an important concept in Chinese medicine, and many herbs possess both therapeutic and health-preserving functions, a fact widely documented and applied in ancient Chinese herbal medicine texts.

[0006] Li Ruiguo et al. added a compound of traditional Chinese medicines, including peppermint, dandelion, houttuynia cordata, honeysuckle, and isatis root, to canine chewing food. The compound did not have an adverse effect on the palatability of the food, but it could regulate the intestinal flora structure of dogs and affect the levels of triglycerides, albumin and catalase to a certain extent (Li Ruiguo, Yang Jie, Shi Baiguan, Li Ronglin, Wang Jianguo, Cai Cuifang, Xin Guosheng. Effects of adding traditional Chinese medicines to chewing food on blood biochemical indicators and intestinal flora in dogs [J]. China Feed, 2022(5):100-106).

[0007] Adding traditional Chinese medicine to pets' joint condition can help prevent disease and address dietary needs for preventative treatment, which requires greater attention. The demand for using traditional Chinese medicine to prevent joint diseases in dogs and cats is increasingly prominent. Summary of the Invention

[0008] To address the problems mentioned in the background section, this invention provides a premix for improving pet joint health and its preparation method. The premix of this invention effectively improves joint pain associated with osteoarthritis, improves the condition of knee joint tissues in osteoarthritis, and has a good preventive and therapeutic effect on osteoarthritis.

[0009] To solve the above-mentioned technical problems, the technical solution provided by the present invention is as follows:

[0010] A premixed feed for improving pet joint health contains the following components in parts by weight: 15-25 parts Polygonatum sibiricum, 15-25 parts Drynaria fortunei, 15-25 parts Eucommia ulmoides, 5-15 parts Ligusticum chuanxiong, 5-15 parts Glycyrrhiza uralensis, 5-15 parts Curcuma longa, 5-15 parts Poria cocos, and 5-15 parts Dioscorea opposita.

[0011] In one embodiment of the present invention, the premix contains the following components in parts by weight: 20 parts of Polygonatum sibiricum, 20 parts of Drynaria fortunei, 20 parts of Eucommia ulmoides, 10 parts of Ligusticum chuanxiong, 10 parts of Glycyrrhiza uralensis, 10 parts of Curcuma longa, 10 parts of Poria cocos, and 10 parts of Dioscorea opposita.

[0012] In one embodiment of the present invention, the premix contains the following components in parts by weight: 25 parts Polygonatum sibiricum, 15 parts Drynaria fortunei, 25 parts Eucommia ulmoides, 5 parts Ligusticum chuanxiong, 15 parts Glycyrrhiza uralensis, 5 parts Curcuma longa, 15 parts Poria cocos, and 5 parts Dioscorea opposita.

[0013] In one embodiment of the present invention, the premix contains the following components in parts by weight: 15 parts Polygonatum sibiricum, 25 parts Drynaria fortunei, 15 parts Eucommia ulmoides, 15 parts Ligusticum chuanxiong, 5 parts Glycyrrhiza uralensis, 15 parts Curcuma longa, 5 parts Poria cocos, and 15 parts Dioscorea opposita.

[0014] The present invention provides a method for preparing the above-mentioned premix, wherein the preparation method is as follows: the prescribed amounts of the traditional Chinese medicinal herbs Polygonatum sibiricum, Drynaria fortunei, Eucommia ulmoides, Ligusticum chuanxiong, Glycyrrhiza uralensis, Curcuma longa, Poria cocos, and Dioscorea opposita are premixed, pulverized, and sieved to obtain a premix that improves the joint health of pets.

[0015] This invention provides the application of the above-mentioned premix for improving pet joint health in the preparation of dog or cat food.

[0016] Furthermore, the premix of the present invention improves joint health in pets and alleviates joint pain in dogs with osteoarthritis.

[0017] The premixed feed for improving pet joint health, as described in this invention, is used at a base diet of 0.8%-5.0%.

[0018] Furthermore, the amount of the premixed feed that improves pet joint health is 2.4% of the base diet.

[0019] The beneficial effects achieved by this invention are as follows:

[0020] This invention's premix contains the following natural ingredients: Polygonatum sibiricum, Drynaria fortunei, Eucommia ulmoides, Ligusticum chuanxiong, Glycyrrhiza uralensis, Curcuma longa, Poria cocos, and Dioscorea opposita. Eucommia ulmoides and Drynaria fortunei are the principal herbs, tonifying the kidneys and strengthening bones, fortifying tendons and activating collaterals, promoting blood circulation and healing injuries, thus strengthening the bone and joint structure from the root. Polygonatum sibiricum and Dioscorea opposita are the assistant herbs, tonifying qi and nourishing yin, strengthening the spleen and kidneys, consolidating the tonifying effect of the principal herbs, and also supporting spleen and stomach function, preventing cloying stagnation. Ligusticum chuanxiong, Curcuma longa, and Poria cocos are the adjuvant herbs, promoting blood circulation and removing dampness, clearing joints, eliminating local congestion, edema, and inflammation in the joints, and relieving pain and other symptoms. Glycyrrhiza uralensis is the guiding herb, harmonizing the effects of all the herbs. The entire formula tonifies the liver and kidneys to strengthen the foundation, promotes blood circulation and clears collaterals to treat the symptoms, strengthens the spleen and removes dampness to aid digestion, tonifies the spleen without causing dampness retention, and eliminates dampness without harming the body's vital energy, treating both the root cause and the symptoms, combining tonification and activating, with the medicinal effects directly reaching the joints.

[0021] Pharmacological studies have shown that the components added to the premix of this invention have a synergistic effect in treating arthritis in pets, effectively improving joint pain in rats with osteoarthritis, improving the condition of knee joint tissues in rats with osteoarthritis, and having a good therapeutic effect on rats with osteoarthritis.

[0022] This invention provides a premixed feed that improves pet joint health by strengthening cartilage, ligaments, relieving pain, and replenishing synovial fluid, effectively improving pet arthritis health and enhancing their quality of life. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in this invention or the prior art, the accompanying drawings used in the specific embodiments will be briefly described below.

[0024] Figure 1 To compare the foot withdrawal thresholds of each group under mechanical stimulation, and to compare with group C, & P < 0.05 && P < 0.01; compared with group M, * P < 0.05 ** P < 0.01; compared with group S1, # P < 0.05 ## P < 0.01.

[0025] Figure 2 The percentage of weight-bearing on the right hind limb was compared among the groups, and compared with group C. & P < 0.05 && P < 0.01; compared with group M, * P < 0.05 ** P < 0.01; compared with group S1, # P < 0.05 ## P < 0.01.

[0026] Figure 3 For comparison of knee joint scores among groups, and comparison with group C, & P < 0.05 &&P < 0.01; compared with group M, * P < 0.05 ** P < 0.01; compared with group S1, # P < 0.05 ## P < 0.01.

[0027] Figure 4 To compare the mechanical pain thresholds of mice in each group, and compared with group C, & P < 0.05 && P < 0.01; compared with group M, * P < 0.05 ** P < 0.01; compared with group S1, # P < 0.05 ## P < 0.01.

[0028] Figure 5 To compare the stride length of mice in each group, and compared with group C, & P < 0.05 && P < 0.01; compared with group M, * P < 0.05 ** P < 0.01; compared with group S1, # P < 0.05 ## P < 0.01.

[0029] Figure 6 To compare the TNF-α levels of mice in each group, and compared with group C, & P < 0.05 && P < 0.01; compared with group M, * P < 0.05 ** P < 0.01; compared with group S1, # P < 0.05 ## P < 0.01.

[0030] Figure 7 To compare the IL-1β levels of mice in each group, and compared with group C, & P < 0.05 && P < 0.01; compared with group M, * P < 0.05 ** P < 0.01; compared with group S1, # P < 0.05 ## P < 0.01.

[0031] Figure 8 To compare the IL-6 levels of mice in each group, and compared with group C, & P < 0.05 && P < 0.01; compared with group M, * P < 0.05** P < 0.01; compared with group S1, # P < 0.05 ## P < 0.01. Detailed Implementation

[0032] The present invention is illustrated below with specific embodiments, but these are not intended to limit the scope of the invention. The described embodiments are merely some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0033] As used herein, the terms “prepared from” and “comprising” are synonymous. The terms “comprising,” “including,” “having,” “containing,” or any other variations thereof, as used herein, are intended to cover non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such composition, step, method, article, or apparatus.

[0034] When a quantity, concentration, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper and lower preferred values, it should be understood as specifically disclosing all ranges formed by any pair of any upper or preferred value with any lower or preferred value, regardless of whether the range is disclosed individually.

[0035] Where no specific technology or conditions are specified in the embodiments, the technology or conditions described in the literature in this field, or the product manual, shall be followed.

[0036] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; unless otherwise specified, the experimental materials and raw materials used in the following examples are all purchased from commercial channels.

[0037] Example 1

[0038] A premixed feed for improving pet joint health contains the following components in parts by weight: 20 parts Polygonatum sibiricum, 20 parts Drynaria fortunei, 20 parts Eucommia ulmoides, 10 parts Ligusticum chuanxiong, 10 parts Glycyrrhiza uralensis, 10 parts Curcuma longa, 10 parts Poria cocos, and 10 parts Dioscorea opposita.

[0039] The preparation method includes the following steps: premixing the prescribed amounts of Chinese herbal medicines such as Polygonatum sibiricum, Drynaria fortunei, Eucommia ulmoides, Ligusticum chuanxiong, Glycyrrhiza uralensis, Curcuma longa, Poria cocos, and Dioscorea opposita, then pulverizing and sieving them to obtain a premixed feed that improves the joint health of pets.

[0040] Example 2

[0041] A premixed feed for improving pet joint health contains the following components in parts by weight: 20 parts Polygonatum sibiricum, 20 parts Drynaria fortunei, 20 parts Eucommia ulmoides, 10 parts Ligusticum chuanxiong, 10 parts Glycyrrhiza uralensis, 10 parts Curcuma longa, 10 parts Poria cocos, and 10 parts Dioscorea opposita.

[0042] The preparation method is the same as in Example 1.

[0043] Example 3

[0044] A premixed feed for improving pet joint health contains the following components in parts by weight: 15 parts Polygonatum sibiricum, 25 parts Drynaria fortunei, 15 parts Eucommia ulmoides, 15 parts Ligusticum chuanxiong, 5 parts Glycyrrhiza uralensis, 15 parts Curcuma longa, 5 parts Poria cocos, and 15 parts Dioscorea opposita.

[0045] The preparation method is the same as in Example 1.

[0046] Example 4

[0047] A premixed feed for improving pet joint health contains the following components in parts by weight: 12 parts Polygonatum sibiricum, 20 parts Drynaria fortunei, 12 parts Eucommia ulmoides, 10 parts Ligusticum chuanxiong, 7 parts Glycyrrhiza uralensis, 12 parts Curcuma longa, 7 parts Poria cocos, and 17 parts Dioscorea opposita.

[0048] The preparation method is the same as in Example 1.

[0049] Example 5

[0050] A premixed feed for improving pet joint health contains the following components in parts by weight: 18 parts Polygonatum sibiricum, 18 parts Drynaria fortunei, 15 parts Eucommia ulmoides, 15 parts Ligusticum chuanxiong, 5 parts Glycyrrhiza uralensis, 15 parts Curcuma longa, 5 parts Poria cocos, and 15 parts Dioscorea opposita.

[0051] The preparation method is the same as in Example 1.

[0052] Example 6

[0053] A premixed feed for improving pet joint health contains the following components in parts by weight: 15 parts Polygonatum sibiricum, 25 parts Drynaria fortunei, 15 parts Eucommia ulmoides, 5 parts Ligusticum chuanxiong, 12 parts Glycyrrhiza uralensis, 18 parts Curcuma longa, 7 parts Poria cocos, and 10 parts Dioscorea opposita.

[0054] The preparation method is the same as in Example 1.

[0055] Comparative Example 1

[0056] A premixed feed for improving pet joint health contains the following components in parts by weight: 20 parts Polygonatum sibiricum, 20 parts Eucommia ulmoides, 10 parts Ligusticum chuanxiong, 10 parts Glycyrrhiza uralensis, 10 parts Curcuma longa, 10 parts Poria cocos, and 10 parts Dioscorea opposita.

[0057] Compared with Example 1, the difference is that it does not contain Drynaria fortunei, while the other components and dosages remain the same.

[0058] The preparation method is the same as in Example 1.

[0059] Comparative Example 2

[0060] A premixed feed for improving pet joint health contains the following components in parts by weight: 20 parts Polygonatum sibiricum, 20 parts Drynaria fortunei, 10 parts Ligusticum chuanxiong, 10 parts Glycyrrhiza uralensis, 10 parts Curcuma longa, 10 parts Poria cocos, and 10 parts Dioscorea opposita.

[0061] Compared with Example 1, the difference is that it does not contain Eucommia ulmoides, while the other components and dosages remain the same.

[0062] The preparation method is the same as in Example 1.

[0063] Comparative Example 3

[0064] A premixed feed for improving pet joint health contains the following components in parts by weight: 20 parts Drynaria fortunei, 20 parts Eucommia ulmoides, 10 parts Ligusticum chuanxiong, 10 parts Glycyrrhiza uralensis, 10 parts Curcuma longa, 10 parts Poria cocos, and 10 parts Dioscorea opposita.

[0065] Compared with Example 1, the difference is that it does not contain Polygonatum sibiricum, while the other components and their amounts remain the same.

[0066] The preparation method is the same as in Example 1.

[0067] Comparative Example 4

[0068] A premixed feed for improving pet joint health contains the following components in parts by weight: 20 parts Polygonatum sibiricum, 20 parts Drynaria fortunei, 20 parts Eucommia ulmoides, 10 parts Glycyrrhiza uralensis, 10 parts Curcuma longa, 10 parts Poria cocos, and 10 parts Dioscorea opposita.

[0069] Compared with Example 1, the difference is that it does not contain Ligusticum chuanxiong, while the other components and dosages remain the same.

[0070] The preparation method is the same as in Example 1.

[0071] Comparative Example 5

[0072] A premixed feed for improving pet joint health contains the following components in parts by weight: 20 parts Polygonatum sibiricum, 20 parts Dipsacus asper, 20 parts Eucommia ulmoides, 10 parts Ligusticum chuanxiong, 10 parts Glycyrrhiza uralensis, 10 parts Curcuma longa, 10 parts Poria cocos, and 10 parts Dioscorea opposita.

[0073] Compared with Example 1, the difference is that Dipsacus is used instead of Drynaria fortunei, while the other components and dosages remain the same.

[0074] The preparation method is the same as in Example 1.

[0075] Comparative Example 6

[0076] The formulation in Example 1 of Chinese Patent CN110585263A (A joint ointment and its preparation method).

[0077] It contains the following components: Drynaria fortunei, Lycopodium clavatum, tortoise shell glue, Eucommia ulmoides, Cynanchum paniculatum, Psoralea corylifolia extract, glucosamine hydrochloride, chondroitin sulfate, and green-lipped mussel powder; and the weights of the above components are as follows: Drynaria fortunei 15g, Lycopodium clavatum 15g, tortoise shell glue 10g, Eucommia ulmoides 15g, Cynanchum paniculatum 15g, Psoralea corylifolia extract 15g, glucosamine hydrochloride 20g, chondroitin sulfate 12g, and green-lipped mussel powder 5g.

[0078] The product was obtained by following the preparation method described in the patent.

[0079] Study on the effect of the premix of this invention on osteoarthritis in rats

[0080] 1. Experimental Methods

[0081] Sixty SPF-grade male SD rats, weighing 180-220g, were randomly selected as follows: 6 rats were assigned to the sham-operated group, and the remaining rats were assigned to the model group. Rats in the sham-operated group were injected intra-articularly with 0.9% saline, while rats in the other groups were injected with sodium iodoacetate to establish an osteoarthritis model.

[0082] The specific procedure was as follows: After anesthetizing the rats, the hair in the 1cm area around the knee joint of the right hind limb was shaved, and the area was disinfected. With the knee joint flexed at 90 degrees, the joint cavity was punctured from the lateral side of the subosseous ligament, with the needle inserted at a 90-degree angle to the tibia, and 50μL of sodium iodoacetate was injected. After the rats injected with sodium iodoacetate extended and flexed their right hind limb 5 times, they were returned to their cages and allowed to move freely for 14 days. The sham-operated group was injected with 50μL of 0.9% physiological saline in the same manner.

[0083] After modeling, the rats were randomly divided into four groups: M, S1, S2, S3, T1, T2, T3, T4, and T5, with six rats in each group.

[0084] The experimental groups are shown below: Group C was the sham surgery group, fed with a basal diet; Group M served as the model control group, fed with a basal diet. Group S1 was fed a basal diet supplemented with 0.96% of the premix from Example 1; Group S2 was fed a basal diet supplemented with 0.96% of the premix from Example 2; Group S3 was fed a basal diet supplemented with 0.96% of the premix from Example 3; Group T1 was supplemented with 0.96% of Comparative Example 1 premix in its basal diet; Group T2 was supplemented with 0.96% of Comparative Example 2 premix in its basal diet; Group T3 was supplemented with 0.96% of Comparative Example 3 premix in its basal diet; The T4 group was supplemented with 0.96% of the Comparative Example 4 premix in the basal diet; The T5 group had 0.96% of Comparative Example 5 premix added to their basal diet.

[0085] The basal diets for rats in each group were purchased from Keao Cooperation (Tianjin) Feed Co., Ltd.

[0086] Feeding began on day 14 after successful model establishment and continued for 28 days.

[0087] 2. Detection indicators

[0088] 2.1 Mechanical pain threshold measurement

[0089] On day 28 after modeling, the mechanical pain threshold of the right hind limb of rats was detected using an electronic Von Frey pain meter.

[0090] The center of the toe surface of the rat's right hind limb paw was stimulated with Von Frey fibers, and pressure was applied vertically and evenly upward. When the rat's hind limb exhibited a paw retraction, paw lifting, or paw licking response, a measurement was completed. The pain meter automatically recorded the maximum value detected. The measurement was repeated 3 times, and the average value was taken as the mechanical stimulation paw retraction threshold.

[0091] 2.2 Load-bearing test

[0092] On day 28 post-modeling, weight-bearing on both feet was assessed using a bipedal balance tester. The degree of pain in the rat limbs was evaluated by measuring the weight-bearing distribution on the hind limbs of the modeling side and the blank control side.

[0093] The rat's forelimbs were placed on the restraint plate, and its two hind limbs were placed on two high-sensitivity weight test plates. The positions of both sides were kept consistent, and the average weight-bearing of the left and right hind limbs was recorded. Three measurements were taken and the average value was calculated to calculate the percentage of weight-bearing on the right hind limb.

[0094] The percentage of weight-bearing on the right hind limb = weight of the right hind limb / (weight of the left hind limb + weight of the right hind limb) × 100%.

[0095] 2.3 Gross observation of the knee joint

[0096] After administration, the rats were euthanized, and the right knee joint was dissected to fully expose the distal femoral articular surface, articular groove, and internal and external surfaces. The articular cartilage, synovial tissue, and joint capsule were observed, recorded, and scored.

[0097] The scoring criteria are as follows: A smooth joint surface with a light blue, semi-transparent appearance is scored as 0 points.

[0098] The joint surfaces are slightly rough or have small cracks, and the color is dull, which is scored as 1 point;

[0099] Rough articular surfaces with visible cartilage defects extending to the middle layer of cartilage are scored as 2 points.

[0100] The joint surface is extremely rough, with ulceration and cartilage defects extending to the deep cartilage layer, which is scored as 3 points;

[0101] Extensive loss and detachment of articular cartilage, with exposure of subchondral bone, is scored as 4 points.

[0102] 3. Statistical Methods

[0103] Data processing was performed using GraphPad Prism 7.0 software, with the measurement data presented in... ±s indicates that the comparison between groups was performed using one-way ANOVA, and P<0.05 was considered statistically significant.

[0104] 4. Experimental Results

[0105] 4.1 Effects on the foot withdrawal threshold of rats under mechanical stimulation

[0106] Table 1 Results of foot withdrawal thresholds for each group under mechanical stimulation

[0107] Note: Compared with C, & P < 0.05 && P < 0.01; Compared with M, * P < 0.05 ** P < 0.01; Compared with group S1, # P < 0.05 ## P < 0.01.

[0108] The results are shown in Table 1 and... Figure 1 .

[0109] Compared with group C, rats in group M had a significantly lower paw withdrawal threshold under mechanical stimulation.

[0110] Compared with group M, rats in groups S1, S2 and S3 showed a significant increase in the mechanical stimulation threshold for foot withdrawal.

[0111] Compared with groups T1-T5, the groups in the examples showed significant differences in increasing the mechanical stimulation threshold for foot withdrawal.

[0112] 4.2 Effect on the percentage of weight-bearing limb of the right hind limb in rats

[0113] Table 2 Results of weight-bearing percentage of the right hind limb in each group

[0114] Note: Compared with C, & P < 0.05 && P < 0.01; Compared with M, * P < 0.05 **P < 0.01; Compared with group S1, # P < 0.05 ## P < 0.01.

[0115] The results are shown in Table 2 and Figure 2 .

[0116] Compared with group C, the percentage of weight-bearing on the right hind limb of rats in group M was significantly reduced.

[0117] Compared with group M, the percentage of weight-bearing on the right hind limb of rats in groups S1, S2 and S3 was significantly increased.

[0118] Compared with groups T1-T5, the groups in the examples showed significant differences in the percentage increase in weight-bearing on the right hind limb.

[0119] 4.3 Effects on rat knee joint scores

[0120] Table 3 Knee joint score results for each group

[0121] Note: Compared with C, & P < 0.05 && P < 0.01; Compared with M, * P < 0.05 ** P < 0.01; Compared with group S1, # P < 0.05 ## P < 0.01.

[0122] The results are shown in Table 3 and Figure 3 .

[0123] Compared with group C, the knee joint score of rats in group M was significantly increased.

[0124] Compared with group M, the knee joint scores of rats in groups S1, S2 and S3 were significantly lower.

[0125] Compared with groups T1-T5, the groups in the examples showed significant differences in reducing knee joint scores.

[0126] The results show that the premixes in Examples 1-3 have a good effect on improving knee osteoarthritis. Comparative Examples 1-5 show that reducing the number of components and changing the composition resulted in varying degrees of decreased improvement. Comparing Examples 1-3 with Comparative Examples 1-5, it was found that the addition of Drynaria fortunei, Eucommia ulmoides, and Polygonatum sibiricum helps to enhance the efficacy, and the combined effect of these three ingredients further enhances the therapeutic function of the composition in treating osteoarthritis.

[0127] By measuring the mechanical pain threshold and weight-bearing distribution in both feet of osteoarthritis rats, it was found that the premix of this invention improved joint pain in osteoarthritis rats. Knee joint scoring in osteoarthritis rats showed that the premix of this invention effectively improved knee joint tissue condition. The premix of this invention has a good therapeutic effect on osteoarthritis rats.

[0128] Study on the effect of the premix of this invention on osteoarthritis in mice

[0129] 1. Experimental Methods

[0130] Thirty-two SPF-grade C57BL / 6J mice, weighing 18-22g, were randomly selected as the sham-operated group (Group C), with the remaining mice serving as the model group. Except for the sham-operated group, all other groups underwent medial meniscus instability surgery to establish a knee osteoarthritis model in the right hind limb. In the sham-operated group, the skin of the right hind limb was incised, the knee joint was exposed, and then sutured. After modeling, the mice were randomly divided into three groups: M, S1, and T6, with eight mice in each group.

[0131] The experimental groups are shown below:

[0132] Group C was the sham-operated group, fed a basal diet; Group M was the model control group, fed a basal diet; Group S1 had 0.87% of Example 1 premix added to the basal diet; Group T6 had 0.87% of Comparative Example 6 product added to the basal diet. The basal diets for mice in each group were purchased from Keao Cooperation (Tianjin) Feed Co., Ltd.

[0133] Starting from the second day after modeling, the above composition was fed continuously for 28 days.

[0134] 2. Detection indicators

[0135] 2.1 Mechanical pain threshold measurement

[0136] After the last administration, the 50% paw withdrawal threshold of mice was determined using an electronic Von Frey pain meter.

[0137] 2.2 Measurement of mouse gait

[0138] The stride length of mice was determined and analyzed using footprint analysis. The stride path was 50 cm long and 7 cm wide. The stride length was the straight-line distance between two consecutive footprints. The stride length was measured three times and the average value was taken as the stride length value.

[0139] 2.3 Detection of serum inflammatory factor levels

[0140] The levels of inflammatory factors tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and interleukin 6 (IL-6) in serum were detected by ELISA.

[0141] 3. Statistical Methods

[0142] Data processing was performed using GraphPad Prism 7.0 software, with the measurement data presented in... ±s indicates that the comparison between groups was performed using one-way ANOVA, and P<0.05 was considered statistically significant.

[0143] 4. Experimental Results

[0144] 4.1 Effects on the mechanical pain threshold in mice

[0145] See results Figure 4 .

[0146] Compared with group C, mice in group M showed a significantly lower 50% withdrawal threshold. Compared with group M, mice in group S1 showed a significantly higher 50% withdrawal threshold. Compared with group T6, group S1 showed a significant difference in increasing the 50% withdrawal threshold.

[0147] 4.2 Effects on mouse gait See results Figure 5 .

[0148] Compared to group C, mice in group M showed a significantly reduced stride length. Compared to group M, mice in group S1 showed a significantly increased stride length. Compared to group T6, group S1 showed a significant difference in increasing stride length.

[0149] 4.3 Effects on serum inflammatory factors in mice See results Figure 6 , Figure 7 , Figure 8 .

[0150] Compared with group C, the serum levels of TNF-α, IL-1β, and IL-6 in group M mice were significantly increased. Compared with group M, the serum levels of TNF-α, IL-1β, and IL-6 in group S1 mice were significantly decreased. Compared with group T6, group S1 showed a significant difference in reducing serum levels of TNF-α, IL-1β, and IL-6 in mice.

[0151] The results above show that the premix of Example 1 has a better effect on improving knee osteoarthritis compared to the product of Comparative Example 6.

[0152] In summary, the premix of this invention may achieve the effect of treating knee osteoarthritis through anti-oxidation, and achieves better technical results compared with existing products.

[0153] Actual feeding test of the premix of this invention

[0154] 1. Laboratory animals

[0155] This experiment used 20 dogs. All animals were at least 5 years old. None of the animals had a history of immune-mediated diseases, were allergic, or had other diseases that could cause chronic gastrointestinal dysfunction, including liver disease, pancreatic insufficiency, metabolic diseases, parasitic diseases, and kidney diseases. The animals had not received any medications or diets related to the tested function within the past three months, nor had they used antibiotics, immunosuppressive drugs, or undergone surgery. Animals that were pregnant, lactating, or unable to be fed orally or by gavage were excluded.

[0156] 2. Experimental Design

[0157] The experiment consisted of two treatment groups, with 10 dogs in each group: 1) Control group: fed a basal diet; 2) Experimental group: fed a basal diet supplemented with 2.4% of the premix from Example 1. The basal diets for each group were purchased from Keao Cooperation (Tianjin) Feed Co., Ltd.

[0158] The formal experimental period lasted 42 days. On day 42, 4 mL of blood was collected from each animal via a 10 mL vacuum blood collection tube through a vein in its forelimb. 2 mL of this blood was added to a non-anticoagulated tube, allowed to stand for 30 minutes, and the supernatant was collected and centrifuged; the supernatant was the serum and stored at low temperature. The other 2 mL of blood was added to an anticoagulant tube, the anticoagulant and blood were mixed, and the supernatant was collected after centrifugation; this was the plasma and stored at low temperature. On days 0 and 42 of the formal experimental period, lameness scores, body weight, and X-ray assessments were performed on all 20 experimental animals.

[0159] 3. Detection Indicators and Methods

[0160] 3.1 Inflammatory markers

[0161] Inflammatory markers included tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 3 (MMP-3), matrix metalloproteinase 9 (MMP-9), and matrix metalloproteinase 13 (MMP-13). All inflammatory markers were measured using enzyme-linked immunosorbent assay (ELISA) according to the instructions of the commercially available kits.

[0162] 3.2 Limping score

[0163] The condition of the dog's joints was scored according to the clinical lameness scoring criteria for dogs, as shown in Table 4.

[0164] Table 4. Clinical lameness scoring criteria for dogs

[0165] 4. Statistical Analysis

[0166] Using SAS 9.4 software (SASInst.Inc.,Cary,NC,USA), with each experimental animal as the experimental unit, the data were first tested for normality. After confirming that the data met the normal distribution, independent samples were then processed. t If the distribution does not conform to a normal distribution, a nonparametric test is performed. Results are expressed as mean ± standard error of mean, and statistical results are expressed in terms of... P <0.05 was considered statistically significant, and 0.05 ≤ P <0.10 indicates a divergent trend.

[0167] 5. Results and Analysis

[0168] 5.1 Inflammatory markers

[0169] The results of the inflammatory marker tests are shown in Table 5.

[0170] Table 5 Results of Inflammatory Marker Detection

[0171] On day 42 of the experiment, compared with the control group, the serum levels of IL-6 and MMP-13 in the experimental group were significantly lower. P <0.05), and TNF-α levels showed a decreasing trend (0.05≤ P ≤0.10).

[0172] This indicates that the test sample was able to alleviate the joint inflammation in dogs to some extent.

[0173] 5.2 Limping score

[0174] The results of the canine clinical lameness score are shown in Table 6.

[0175] Table 6 Limping Score Results

[0176] The results showed that the clinical lameness score of the experimental group dogs decreased on day 42, and the decrease was more significant compared with the control group.

[0177] TNF-α, as a core pro-inflammatory cytokine, can activate synovial cells to release more inflammatory mediators, accelerate cartilage matrix degradation, promote osteoclast activation, and also induce systemic discomfort. IL-1β is a typical primary inflammatory cytokine that can activate complex signaling pathways, including cytokines, chemokines, and adhesion factors, playing a crucial role in the initiation and coordination of inflammation. IL-1β not only inhibits the production of type II collagen and reduces the formation of cartilage matrix (such as proteoglycans and collagen), but also strongly stimulates the release of MMPs, leading to chondrocyte degradation, cartilage matrix destruction, and cartilage structure damage. IL-6 can stimulate inflammatory cells to release other pro-inflammatory factors, exacerbating the inflammatory response. Matrix metalloproteinases (MMPs) are enzymes that degrade connective tissue and matrix proteins, playing important regulatory roles in vivo. However, abnormal activation or overexpression of MMPs can lead to articular cartilage destruction and joint diseases. Articular cartilage lysis refers to the process of articular cartilage destruction and degradation, and is an important factor causing joint diseases such as osteoarthritis. Among them, MMP-2 mainly degrades type IV, V, VII, and X collagen, MMP-3 mainly participates in collagen degradation, MMP-9 has the core function of degrading type IV collagen, and MMP-13 specifically degrades type II collagen. Elevated serum MMP-13 is associated with the destruction of articular cartilage and the development of osteoarthritis.

[0178] Under the conditions of this experiment, on day 42, compared with the control group, feeding with the premix of Example 1 significantly reduced the levels of interleukin-6 and matrix metalloproteinase 13 in canine serum (P<0.05), while also showing a decreasing trend in the level of tumor necrosis factor-α in canine serum (0.05≤P≤0.10). This indicates that the premix of Example 1 can alleviate joint inflammation in dogs to some extent.

[0179] Under the conditions of this experiment, the results showed that the clinical lameness score of the experimental group dogs decreased on day 42, and the decrease was more significant compared with the control group. This indicates that the premix of Example 1 improved the lameness of the dogs to some extent.

[0180] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, it is intended that all variations falling within the meaning and scope of equivalents of the claims be included within the present invention.

Claims

1. A premix for improving joint health in pets, characterized in that, The premix contains the following components in parts by weight: 15-25 parts of polygonatum, 15-25 parts of drynaria, 15-25 parts of eucommia, 5-15 parts of chuanqiong, 5-15 parts of licorice, 5-15 parts of curcuma, 5-15 parts of poria, and 5-15 parts of yam.

2. The premix for improving joint health of pets according to claim 1, characterized in that, The premix contains the following components in parts by weight: 20 parts of polygonatum, 20 parts of drynaria, 20 parts of eucommia, 10 parts of chuanqiong, 10 parts of licorice, 10 parts of curcuma, 10 parts of poria, and 10 parts of yam.

3. The premix for improving joint health of pets according to claim 1, wherein The premix contains the following components in parts by weight: 25 parts of polygonatum, 15 parts of drynaria, 25 parts of eucommia, 5 parts of chuanqiong, 15 parts of licorice, 5 parts of curcuma, 15 parts of poria, and 5 parts of yam.

4. The premix for improving joint health of pets according to claim 1, wherein, The premix contains the following components in parts by weight: 15 parts of polygonatum, 25 parts of drynaria, 15 parts of eucommia, 15 parts of chuanqiong, 5 parts of licorice, 15 parts of curcuma, 5 parts of poria, and 15 parts of yam.

5. The pre-mix for improving joint health of pets according to claim 1, wherein, The preparation method comprises the following steps: mixing the Chinese herbal medicines polygonatum, drynaria, eucommia, chuanqiong, licorice, curcuma, poria, and yam in formula amount, crushing, and sieving to obtain the premix for improving joint health of pets.

6. Use of the premix for improving joint health of pets according to any one of claims 1-4 in the preparation of dog or cat feed.

7. Use according to claim 6, characterized in that, The premix improves joint pain of dog or cat osteoarthritis.

8. Use according to claim 6, characterized in that, The use amount of the premix for improving joint health of pets is 0.8%-5.0% of the basal daily ration.

9. Use according to claim 8, characterized in that, The use amount of the premix for improving joint health of pets is 2.4% of the basal daily ration.

Images