WO2023197407A1

WO2023197407A1 - Use of hugu capsule in preparation of a drug for treating steroid-induced osteonecrosis of the femoral head

Info

Publication number: WO2023197407A1
Application number: PCT/CN2022/094928
Authority: WO
Inventors: 潘翠萍; 黄慧云; 方淑玲; 王春彬
Original assignee: 广东安诺药业股份有限公司
Priority date: 2022-04-11
Filing date: 2022-05-25
Publication date: 2023-10-19
Also published as: CN114601892A

Abstract

Disclosed is use of a HuGu capsule in the preparation of a drug for treating steroid-induced osteonecrosis of the femoral head. The HuGu capsule is prepared from radix polygoni multiflori preparata, epimedium, radix rehmanniae praeparata, tortoise shell, morinda officinali, eucommia ulmoides, radix dipsaci, rhizoma drynariae, angelica sinensis, and Chinese yam. It is proved in the present invention that the drug can inhibit the high-conversion state of bone metabolism in a model of the steroid-induced osteonecrosis of the femoral head, inhibit bone resorption, improve the internal environment of the femoral head, and regulate the bone metabolism, providing an experimental basis for the treatment of the steroid-induced osteonecrosis of the femoral head.

Description

Use of Hugu Capsules in the preparation of drugs for the treatment of steroid-induced femoral head necrosis

Technical field

The invention belongs to the technical field of traditional Chinese medicine application, and specifically relates to the use of a bone-protecting capsule in preparing medicine for treating steroid-induced femoral head necrosis.

Background technique

Since Pietrogrande and Mastromarino first reported the case of femoral head necrosis caused by the use of glucocorticoids in 1957, steroid-induced femoral head necrosis has gradually attracted widespread attention from the medical community at home and abroad. Its incidence rate has increased year by year. It is the most common cause of femoral head necrosis following trauma. The second largest reason. Hormone-induced avascular necrosis of the femoral head is called bone erosion in traditional Chinese medicine. Traditional Chinese medicine believes that when righteousness is deficient, the three qi of wind, cold, and dampness invade the human body and cause paralysis. This has been recorded by doctors of the past dynasties, "Su Wen·Bi Lun" It was pointed out that there are many reasons for the formation of paralysis, but the key factors in its pathogenesis are qi stagnation and blood stasis, disharmony of the veins, and liver and kidney insufficiency.

The complete mechanism of glucocorticoid-induced femoral head necrosis is not clear yet. Currently, steroid-induced femoral head necrosis is more likely to be caused by fat embolism, blood coagulation disorder, thrombosis, insufficient blood supply, regulation of autophagy and apoptosis, etc. The combined effect is to cause the apoptosis of bone cells and osteoblasts, prolong the life span of osteoclasts, and cause changes in trabecular bone, which may lead to osteoporosis, decreased bone hardness, and prone to femoral head necrosis.

HuGu capsule (HG) is a traditional Chinese medicine that nourishes the kidneys, strengthens the spleen, activates blood circulation and removes blood stasis. The formula is: Polygonum multiflorum, Epimedium, Rehmannia glutinosa, Turtle shell, Morinda officinalis, Eucommia ulmoides, Dipsacus bark, Drynariae. , angelica, yam. Hugu Capsule has currently proven its therapeutic effect in preventing secondary osteoporosis and traumatic fractures; the entire prescription of Hugu Capsule is mainly to nourish the kidneys and strengthen bones, and there is still room for exploration into other orthopedic diseases.

Contents of the invention

In view of this, the present invention uses a rat model of steroid-induced femoral head necrosis to observe the effect of bone-protecting capsules on the morphology of necrotic femoral head and bone metabolism biochemical indicators, providing an experimental basis for the treatment of steroid-induced femoral head necrosis and expanding the scope of bone-protecting capsules. New indications for capsules.

In order to achieve the above-mentioned objects of the invention, the technical solutions of the present invention are as follows:

Hugu Capsule is used as a drug for the treatment of steroid-induced femoral head necrosis. The traditional Chinese medicine is composed of Polygonum multiflorum, Epimedium, Rehmannia glutinosa, Turtle shell, Morinda officinalis, Eucommia ulmoides, Dipsacus bark, Drynariae, Angelica sinensis and Chinese yam. The application of Hugu Capsule as a drug for the treatment of steroid-induced femoral head necrosis.

In particular, Hugu Capsule inhibits the high turnover state of bone metabolism in steroid-induced femoral head necrosis.

In particular, Hugu Capsule inhibits bone resorption in steroid-induced femoral head necrosis.

In particular, bone-protecting capsules improve the internal environment of the femoral head.

Compared with the prior art, the positive and beneficial effects of the present invention are:

Unlike traumatic fractures, secondary osteoporosis and other diseases that have a clear single mechanism, steroid-induced femoral head necrosis is multi-factorial. Based on the characteristics of steroid-induced femoral head necrosis, the present invention discloses for the first time that bone-protecting capsules can inhibit steroid-induced femoral head necrosis. The high turnover state of bone metabolism in the osteonecrosis model inhibits bone resorption, improves the internal environment of the femoral head and regulates bone metabolism; it provides experimental basis for the drug to be used in the treatment of steroid-induced femoral head necrosis.

Femoral head necrosis is a pathological process in which different causes disrupt the blood circulation of the femoral head, causing necrosis of bone cells, bone marrow hematopoietic cells, and fat cells, ultimately leading to collapse of the femoral head. The pathogenesis of steroid-induced femoral head necrosis is complex and has not yet been fully understood. Its pathological process is complex. If timely and effective treatment is not obtained in the early stage, the cartilage surface will collapse, the joint space will narrow, and eventually osteoarthritis will occur, causing the patient to develop hip disease. Disability due to joint dysfunction. While surgical treatment is often invasive and lacks specific Western medicine treatments, traditional Chinese medicine has shown promising prospects in experimental and clinical research. Traditional Chinese medicine treatment mainly uses traditional Chinese medicine to regulate the movement of qi and blood throughout the body, dredge meridians, and assist with expectoration. It has overall therapeutic effects such as removing dampness, replenishing the liver and kidneys, etc., thereby achieving the purpose of relieving pain, improving function, and promoting necrosis repair.

Currently, the commonly used clinical treatments for steroid-induced femoral head necrosis include methods to preserve the patient's own femoral head and artificial joint replacement. Treatment methods that preserve the patient's own femoral head are divided into non-surgical treatments and surgical treatments. Non-surgical treatments include hyperbaric oxygen, magnetic therapy, and drug therapy. Surgical treatments mostly use drilling decompression, vascular transplantation, and free bone grafting. Surgery, osteotomy, artificial joint replacement and other methods have the disadvantages of high surgical risks, high treatment costs, great pain to the patient, and often unsatisfactory results. The use of Western medicines often has serious side effects, requiring the use of other drugs or hormones, which increases the burden on the patient's body. For example, bisphosphonates have been restricted in European and American countries due to their severe kidney damage effects and are not suitable for use. For femoral head necrosis, a disease that requires long-term clinical medication.

Many scholars at home and abroad have conducted a large number of experiments and research on the treatment of steroid-induced femoral head necrosis, which has laid a rich theoretical foundation for clinical research and provided a reliable basis for multi-factor and multi-target treatment. However, there is currently no unified method. At the same time, relevant research still lacks long-term treatment observation, and the understanding of femoral head necrosis is still very limited. In recent years, with the deepening of medical research, many hypotheses have been put forward about steroid-induced femoral head necrosis, but so far no theory can fully explain its pathogenesis. At present, the advantages of using traditional Chinese medicine to treat steroid-induced femoral head necrosis have become more and more prominent. Traditional Chinese medicine can exert its various effects according to the TCM syndrome prescriptions in different pathogenesis of steroid-induced femoral head necrosis. On the one hand, it inhibits the development of steroid-induced femoral head necrosis. The occurrence and development of the disease, on the other hand, promote the reversal of the disease. Traditional Chinese medicine focuses on the relationship between the liver, spleen, and kidneys, and based on the theory of "activating blood, removing blood stasis, and regenerating", it aims to improve blood hyperviscosity, lower blood lipids, regulate lipid metabolism, improve microcirculation, and reduce vascular endothelial cell damage. Damage and inhibit osteoclasts. In the study of preventing and treating patients with early and mid-stage steroid-induced femoral head necrosis, the best treatment results can be achieved. With the support of the advantages of traditional Chinese medicine and its theoretical theories, the development of Hugu Capsules for the prevention and treatment of steroid-induced femoral head necrosis will make further breakthroughs.

Description of the drawings

Figure 1 is a picture of the toluidine blue staining effect of Hugu Capsule on femoral head cartilage in rats with femoral head necrosis.

Figure 2 is a picture of the toluidine blue staining effect of Hugu Capsule on the growth plate cartilage of the femoral head in rats with femoral head necrosis.

Figure 3 is a picture of the toluidine blue staining effect of Hugu Capsule on the subchondral bone of the femoral head in rats with femoral head necrosis.

In the figure, CON refers to the blank group, MET refers to the modeling group, and HG refers to the bone-protecting capsule group.

Detailed ways

The present invention will be further described in detail below with reference to specific examples. The following examples are not used to limit the present invention, but are only used to illustrate the present invention. Unless otherwise specified, the experimental methods used in the following examples are generally in accordance with conventional conditions. Unless otherwise specified, the experimental methods used in the examples are generally in accordance with conventional conditions. Unless otherwise specified, the materials, reagents, etc. used in the following examples are all Available commercially.

Example 1 Effect of Hugu Capsule on femoral head morphology and bone metabolism biochemical indicators in rats with femoral head necrosis

1Materials and methodsMaterials and methods

1.1 Experimental animals

56 SPF grade SD rats (200±20g) were purchased from the Guangdong Provincial Medical Experimental Animal Center (SCXK (Guangdong) 2013-0002) and raised in the Animal Experimental Center of Guangdong Pharmaceutical University (SYXK (Guangdong) 2017-0125). Under standard laboratory conditions, the rats were fed with standard feed, with free access to water, a constant temperature of 20°C-25°C, and a relative humidity of 40%-70%. All rats were fed with standard rodent feed.

1.2 Drugs and reagents

Bone-protecting capsules (Guangdong Annuo Pharmaceutical Co., Ltd., 0.45g each, national drug approval number Z20040124, production batch number 171107), methylprednisolone sodium succinate (Pfizer Pharmaceuticals Co., Ltd., USA); lipopolysaccharide (Shanghai Beyotime Biotech) Technology Co., Ltd.); penicillin sodium for injection (Harbin Pharmaceutical Group Pharmaceutical Company); chloral hydrate (Sinopharm Chemical Reagent Co., Ltd.); xylene, absolute ethanol, sodium chloride, formic acid, formaldehyde (Tianjin Damao Chemical Reagent Factory); paraffin (LeiCa, USA); toluidine blue dye (Beijing Regen Biotechnology)

1.3 Instruments

Paraffin embedding machine (Leica, Germany, model EG1160); tissue microtome (Leica, Germany, model RM2255); Bioquant-Osteo bone morphology image analysis system (BIOQUANT USA).

1.4 Mold selection method and grouping

Fifty-six SPF adult SD rats, regardless of gender, were fed standard food and had free access to water. After adapting to the experimental environment for one week, they were divided into a blank group of 4 rats and the remaining 52 rats using a random number table method. The blank group was only given the same frequency and dose of normal saline during the modeling period in the following weeks. The remaining 52 animals received two consecutive intraperitoneal injections of lipopolysaccharide (LPS) at a dosage of 20ug/kg, and the two interventions were separated by 24 hours. 24 hours after completing the LPS injection, methylprednisolone sodium succinate (methylprednisolone) 40 mg/kg was injected through the gluteal muscle for three consecutive injections, with an interval of 24 hours between each time.

From the 2nd to 6th week of the experiment, the dosage of methylprednisolone was reduced to 20 mg/kg, once a week, and continued for 5 weeks.

52 SD rats were injected with 80,000 units of penicillin sodium into the gluteal muscle after each modeling session, and were suspended to drink water during drug intervention.

After 6 weeks, 4 rats were randomly selected from the 52 rats as the MOD group, and the 4 rats in the CON group were anesthetized with 10% chloral hydrate intraperitoneally. After anesthesia, fresh femoral heads of SD rats were taken out. The general morphology and light microscope observation of the tissue were performed, and the modeling results were tested.

1.5 Intervention methods

After the model was successfully created, the remaining 48 animals in the experimental group were divided into three groups according to the random number table, the normal saline group, the MET group, and the two bone-protecting capsule dosage groups (according to the conversion of the dosage of human and rat body surface areas, each animal was calculated The dosage of Guhu Capsule required by rats was given as the basic dose and the double dose therapeutic amount of the water extract of Guhu Capsule, which were marked as HG1 group and HG2 group respectively).

Normal saline group The NS group was given 5g/kg/d normal saline every day; the HG1 group and the HG2 group were given two doses of Hugu capsules by gavage respectively, once a day; the rats in the same dosage group were given Before medication, the patients were randomly divided into a 42-day group and a 60-day group, and were administered medication at different times.

1.6 Detection of serum biochemical indicators

After the administration, the rats were fasted for 12 hours and anesthetized intraperitoneally with 10% chloral hydrate 10 mL/kg. After cardiac blood collection, the upper serum was taken. According to the operating instructions of the rat serum enzyme-linked immunoassay kit, the serum bone metabolism of the rats in each group was measured. Biochemical Indicators.

1.7 Morphological staining

After the rats were anesthetized and blood was collected, the right femoral head was removed. After exposing part of the medullary cavity, it was fixed with 4% formaldehyde solution for 24 hours, decalcified with 10% ethylenediaminetetraacetic acid (EDTA) for 2 months, dehydrated with graded alcohol, and cut in the sagittal plane. Open, paraffin-embedded sections, and stained with toluidine blue for morphological comparison.

1.8 Statistical methods

Bioquant-Osteo bone morphology image analysis system software and SPSS 11.0 software were used for data analysis and processing. All data were expressed as mean ± standard deviation (x ± s). One-way analysis of variance and t test were used to compare the data in each group. Statistics A value of P<0.05 indicates that the difference is statistically significant.

2Results

2.1 Modeling process

During the modeling process of rats, the blank group and the CON group were responsive, hyperactive, fed well, had shiny fur, and had regular weight growth. Compared with the CON group, the MET group was slightly slower in response, tired in activity, eating normally, and had loose fur. There was no significant difference between male and female rats.

During the subsequent therapeutic administration, it was observed that the performance of the NS group in the normal saline group did not change compared with that before the administration, while the rats in the two administration groups of Hugu Capsule had better eating and activity responses than before the administration. There has been an improvement, manifested by a relative increase in activity.

In the experiment, there was no collapse in the appearance of the femoral heads in each group. The articular cartilage of the femoral heads in the blank group and CON group was light red in color and had good gloss. When bone scissors were used to extract the materials from the femoral neck, the hardness was relatively hard. When the medullary cavity was exposed for decalcification, there was no bone loss. ; The color of the femoral head articular cartilage in the MET group of the modeling group was slightly white and had good gloss. The bone became obviously brittle when the bone scissors were used to extract materials. When the medullary cavity was exposed for decalcification, the entire articular cartilage was easy to fall off (the shedding rate was 3/8); Therapeutic After administration for a certain period of time, the texture of the femoral heads of rats in the NS group was not significantly improved compared with the MET group; the femoral head articular cartilage of the administration groups HG1 and HG2 was white with red in the middle, with good gloss, and the material was slightly smeared during bone shearing. Brittle, the hardness is between the model group and the blank group.

(1) Surface cartilage area of the femoral head (shown in Figure 1):

In the blank control group, the surface cartilage was evenly stained, the structure was clear, the cartilage surface was smooth, and the structure was continuous and complete. The structure of the transitional layer is obvious, and the immature chondrocytes are mostly flat, with their long axis mostly parallel to the articular surface. The stratum radiatum is deeply stained, and the chondrocytes are round in shape, located in the cartilage lacunae, and arranged in homogeneous cell groups. The tide line is clearly demarcated, and the calcification layer and cartilage cells below the tide line can be seen.

The surface cartilage staining of the model group MET-42d group was lighter, the cartilage surface was less smooth, the transitional layer was less, and the boundary with the radiation layer was unclear. Chondrocytes in the stratum radiata are arranged in homogeneous cell groups, but the cartilage lacunae are mostly empty bone lacunae. There are few or no chondrocytes in the calcified layer below the tide line.

Compared with the model group MET-42d group, the MET-60d group also showed that the cartilage surface was less smooth and the transitional layer was less. However, the chondrocytes in the transitional layer and radial layer had obvious nuclei and few empty bone lacunae. Chondrocytes are rare within the calcified layer.

In the H1-42d group, the basal dose of Hugu Capsules was administered for 42 days. The articular cartilage surface was not smooth, the multi-layered structure of the cartilage was not clear, and the number of chondrocytes increased significantly. The majority of chondrocytes were immature cells, with obvious columnar arrangement and fewer homologous cell groups.

The chondrocytes in the H1-60d group were relatively more mature than those in the 42-day group at the same dose. It can be seen that the cell volume became larger, the cartilage homologous cell group was obvious, and the extracartilage matrix was stained darker.

The H2-42d group of the double-dose Hugu Capsule administration group for 42 days showed that the cartilage structure was still unclear, but the cells were mainly mature chondrocytes. The number of homologous cell groups was significantly increased compared with the H1-42d group, and empty bone cartilage was occasionally seen. trap. In the H2-6Od group, the surface of the articular cartilage was clearly and deeply stained, the fiber bundle structure was continuous and complete, and the structural boundaries between the transitional layer and the radial layer were clearly visible. The cells were still mainly mature chondrocytes, and the number of homologous cell groups was higher than that in the H1-60d group. Increased number, with rare empty bone and cartilage lacunae.

(2) Growth plate area of the femoral head (shown in Figure 2):

After the blank control group was stained with toluidine blue, it was found that the growth plate was darkly stained with toluidine blue, and an obvious three-layer structure was visible, including a quiescent zone, a proliferation zone, and a hypertrophic zone. The quiescent zone is adjacent to the subchondral bone plate above and has a continuous structure, while the chondrocytes in the proliferative and hypertrophic zones are arranged in obvious columns and are adjacent to the cavernous body and trabeculae below.

The model group showed that in the M-42d group, the growth plate in the epiphyseal plate was lightly stained, the growth plate was undulating, and the structure was disordered. The chondrocytes in the resting area gradually decreased, and empty bone lacunae were common. In the model group M-60d, it can be seen that the epiphyseal structure is discontinuous, and there are local horizontal fractures in the adjacent parts of the quiescent zone, the subchondral bone plate above, and the proliferation zone below, which are manifested as structural fractures in the growth plate. There are vacuoles forming in the epiphyseal plate, and the trabeculae below the epiphyseal line become sparse and fractured.

Compared with the model group, on H1-42d in the Hugu Capsule basic dose administration group, cartilage cell nuclei were clearly visible, with a continuous structure and vertically arranged chondrocyte columns, but the hypertrophic chondrocytes were significantly smaller. In the H1-60d group, the growth plate structure was normal and clearly visible. There are many layers of chondrocytes in the quiescent zone connected to the subchondral bone plate, and the cartilage columns in the proliferative zone have a tight structure. The upper and lower structures of the epiphyseal plate are continuous without breakage. Double-dose administration of Hugu Capsules: In the H2-42d group, an increase in vacuoles between the columnar cartilage cells of the growth plate was seen, but this did not affect the continuity of the growth plate and the structure of the upper and lower parts. However, in the H2-60d group, clear growth plates were seen. The structure is multi-layered and continuous, with clear cartilage cells and obvious nuclei.

(3) Subchondral bone area of femoral head (shown in Figure 3):

In the blank control group, the subchondral blood vessels of the femoral head were abundant, and the bone trabeculae were arranged regularly, densely, and plumply. The larger trabecular space was filled with bone marrow tissue. The intramedullary hematopoietic cells were abundant and only a small number of adipocytes were seen. The shape is normal, and the epiphyseal line below the cartilage is continuous and smooth.

Obvious osteonecrosis was seen in the model group M-42d group, which was characterized by disordered arrangement of subchondral bone trabeculae, sparse, narrowed, and broken trabeculae, mixed with a large number of incompletely absorbed trabecular bone fragments, that is, necrotic tissue, reduced number of bone cells, and nuclear Pyknosis, edge gathering, empty bone lacunae increase, hematopoietic tissue in the bone marrow decreases, fat cells enlarge and merge into large vacuoles, the bone marrow is edematous, the lumen is empty, and a large number of thrombosis can be seen in some areas.

The model group M-60d group had significantly weaker osteonecrosis than the M-42d group, the subchondral bone plates were arranged more regularly, the trabecular bone was partially broken, and incompletely absorbed trabecular bone fragments could still be seen, surrounded by connective tissue, and hematopoiesis in the bone marrow Tissue is reduced or even absent, fat cells fuse into large vacuoles, and thrombosis is still seen in intramedullary blood vessels. There is a clear structural break between the subchondral trabeculae and the epiphyseal plate.

The H1-42d group of the 42-day administration group of Hugu Capsules was significantly improved compared to the model group M-42d group. It can be seen that trabecular bone fractures were reduced, and the incompletely absorbed trabecular bone fragments were also significantly reduced, but the arrangement was still disordered. The bone marrow still showed edema and almost no hematopoietic cells, adipocytes fused into vacuoles, and the number and area of thrombus in blood vessels were significantly reduced.

However, the bone trabeculae of the H1-60d group, which was administered at the basal dose for 60 days, were relatively thick and formed a woven network, and the bone resorption activity was weakened. The bone marrow still showed a large number of vacuole-like structures, and the number and area of thrombus in blood vessels were significantly reduced. The underlying epiphyseal plate structure was significantly improved compared with the model group M-60d group, and the complete epiphyseal plate cartilage cell column structure connected to the subchondral bone plate was significantly increased. The disorder of trabecular bone in the H2-42d group after double dose administration of Hugu Capsules for 42 days was improved. The fracture and repair reaction of trabecular bone could be seen. Hematopoietic cells in the bone marrow cavity were still missing, and fat cells were fused and disintegrated. Some thrombi may be seen beneath the calcified cartilage. There is partial structural breakage between the subchondral bone trabeculae and the epiphyseal plate.

However, in the double dose 60-day group H2-60d group, the bone trabecular structure was basically normal, with pyknosis of bone cell nuclei, less edge gathering, and a smaller number of bone lacunae. A clear multi-layered epiphyseal plate structure could be seen, and the structure was continuous. The chondrocytes have a clear shape, obvious nuclei, and a small number of empty bone lacunae. There are abundant subchondral blood vessels and a small amount of thrombus can be seen.

2.2 Effect of Hugu Capsule on bone metabolism indicators in serum of rats with steroid-induced femoral head necrosis

Detect the activities of BALP and TRAP5b, and compare the contents of P1NP and CTX. The results are shown in Tables 1 to 4. Compared with the CON group, there was no significant difference in the activities of BALP and TRAP5b and the concentration of P1NP in the MOD group of the model group, but only the concentration of CTX increased significantly. After intragastric administration of physiological saline for a period of time, it was observed that the activities of BALP and TRAP5b, and the concentrations of P1NP and CTX were significantly increased in the 42-day MET-42d group and the 60-day MET-60d group (all P<0.01), but on the 42-day group There was no difference compared with 60 days; however, after intervention for 42 days and 60 days respectively in the two dose groups of Hugu Capsule, the activities of BALP and TRAP5b, as well as the concentrations of P1NP and CTX were significantly reduced (P<0.01); in the two dosage groups In comparison, the down-regulation of the four indicators in the HG2 group was more significant than that in the HG1 group at both time points (P<0.01); within the same dose administration group, there was no significant difference between days 42 and 60 (P>0.01).

Table 1 Effect of Hugu Capsule on serum BALP activity in rats with steroid-induced femoral head necrosis

Table 2 Effect of Hugu Capsule on serum P1NP concentration in rats with steroid-induced femoral head necrosis

Table 3 Effect of Hugu Capsule on serum TRAP5b concentration in rats with steroid-induced femoral head necrosis

Table 4 Effect of Hugu Capsule on serum TRAP5b concentration in rats with steroid-induced femoral head necrosis

In Table 1-4, compared with the blank control group, #P<0.05, ##P<0.01; compared with the model group at the same point, *P<0.05, **P<0.01; compared with the HG1 group at the same point, △P <0.05, △△P<0.01.

in conclusion

The present invention uses lipopolysaccharide combined with hormone induction method to successfully prepare a rat femoral head avascular necrosis model (ONFH model), which is characterized by osteocyte apoptosis, bone marrow necrosis and bone trabecular fracture. The internal anatomical structure of the femoral head includes articular cartilage and subchondral bone tissue, and its pathological changes will inevitably affect the occurrence and development of femoral head necrosis. Therefore, the present invention conducted morphological observations and discussions on different histological parts of the femoral head.

As shown in Figures 1-3, the toluidine blue staining results of femoral head sections show that the experimental steroid-induced necrosis model was successfully built. The present invention found that in the 60-day MET-60d group of the model group, obvious horizontal fractures occurred in the area where the growth plate appeared. This is considered to be due to the dual factors of growth plate chondrocytes exposed to large doses of hormones and corresponding pathological changes in the femoral head. Under the condition, cell apoptosis and autophagy occur. Even after the hormone is withdrawn, there are still subsequent effects of cell damage, which in turn results in the structural loss of chondrocytes at the growth plate, which in turn weakens the mechanics of the femoral head to withstand longitudinal pressure. Capacity, that is, the stress capacity of the femoral head caused by steroid necrosis is reduced, which can be manifested in that the bone feels obviously brittle when the femur is harvested, and the entire articular cartilage easily falls off when the medullary cavity is exposed for decalcification.

The morphological manifestations of the administration group HG1 and HG2 reflect that Hugu Capsule can effectively improve bone metabolism in steroid-induced femoral head necrosis.

Serum bone biochemical indicators were used to detect bone metabolism. The results showed that compared with the blank group CON group, the blood concentrations of MOD's BALP and P1NP did not change significantly at the end of the modeling. However, after a period of intragastric administration of normal saline, it was found that compared with MOD Compared with the two groups, the concentrations of BALP and P1NP in the MET-42d group and MET-60d group increased significantly (P<0.01), which is closely related to the modeling time and modeling points of lipopolysaccharide combined with hormones. Fluctuations in the concentration of bone metabolism indicators initially appear locally in the bone tissue, manifesting as local concentrations in the bone tissue microenvironment. Only when the concentration reaches a certain range, will it become apparent in systemic blood concentrations. A significant change in animal models of femoral head necrosis is the presence of osteonecrosis and bone resorption. At the end of the modeling, there was no significant change in the blood concentrations of bone formation indicators BALP and P1NP in the MOD group. However, as time went by, the concentrations of BALP and P1NP in the MET-42d group and MET-60d group increased significantly, manifesting as femoral head necrosis. The model has a certain effect on bone formation, but the effect is relatively slow reflected in serum bone metabolism indicators. After the hormone is withdrawn and a period of physiological saline is given, the differentiation activity of osteoblasts increases and bone turnover becomes active; and compared with the model at the same time point Compared with the two groups, after the administration of Hugu Capsule, the concentrations of BALP and P1NP in the two dose groups were significantly decreased (P<0.01), which represents a decrease in the differentiation activity of osteoblasts and a slowdown in bone turnover; at the same time point, the two doses of Comparing the drug groups, BALP and P1NP, which reflect bone turnover and pre-osteoblast differentiation, were more obvious in the HG2 group than in the HG1 group (P<0.01). It can be inferred that Guhugu Capsule can promote bone formation of necrotic femoral head and double bone formation. The dose of bone-protecting capsules used in patients with osteoporosis may be more effective than the basic dose.

Among the indicators reflecting bone resorption, compared with the blank group CON group, the CTX of the modeling group MOD group was significantly increased (P<0.01), while TRACP5b had no significant change, indicating the impact of lipopolysaccharide combined with hormone modeling on bone resorption. It is obvious that the effect is fast, but the effect on osteoclast enzyme activity is limited in a short period of time, which is reflected in the fact that the blood concentration change of TRACP5b is not as obvious as that of CTX; while in the MET group, after intragastric administration of normal saline for a period of time, TRACP5b on day 42 MET- Both the 42d group and the 60-day MET-60d group were significantly elevated, consistent with the performance of CTX, indicating that osteoclasts in the model group were very active, bone resorption accelerated, and type I collagen degradation products increased, which was consistent with the femoral head morphology of the model group. Performance is consistent. Compared with the model group, the CTX and TRACP5b of rats after the administration of Hugu Capsule dropped significantly (P<0.05), indicating that Hugu Capsule can inhibit the activity of rat osteoclasts, inhibit bone resorption, and reduce the degradation products of type I collagen. Reduced; compared with the HG1 group, the two bone resorption indicators of CTX and TRACP5b in the HG2 group decreased more. Therefore, the present invention concluded that bone-protecting capsules can inhibit bone resorption in femoral head necrosis, doubling the amount used in osteoporosis. The dosage of bone-protecting capsules has a better effect on inhibiting the bone resorption of the femoral head.

To sum up, the traditional Chinese medicine compound bone-protecting capsule can inhibit the high turnover state of bone metabolism in the steroid-induced femoral head necrosis model to a certain extent, inhibit bone resorption, improve the internal environment of the femoral head, and regulate bone metabolism. The above effects may be its mechanism for treating steroid-induced femoral head necrosis.

Claims

The application of Hugu Capsule as a drug for treating steroid-induced femoral head necrosis is characterized in that the traditional Chinese medicine is composed of Polygonum multiflorum, Epimedium, Rehmannia glutinosa, Turtle shell, Morinda officinalis, Eucommia ulmoides, Dipsacus and Drynariae. The application of bone-protecting capsules of , angelica and yam as drugs for the treatment of steroid-induced femoral head necrosis.
The application of the bone-protecting capsule according to claim 1 as a drug for treating steroid-induced femoral head necrosis, characterized in that the bone-protecting capsule inhibits the high turnover state of bone metabolism in steroid-induced femoral head necrosis.
The bone-protecting capsule according to claim 1 is used as a drug for treating steroid-induced femoral head necrosis, characterized in that the bone-protecting capsule inhibits bone resorption in steroid-induced femoral head necrosis.
The bone-protecting capsule according to claim 1 is used as a drug for treating steroid-induced femoral head necrosis, characterized in that the bone-protecting capsule improves the internal environment of the femoral head.