CN113117001A - Application of bone-protecting capsule as medicine for treating traumatic fracture - Google Patents
Application of bone-protecting capsule as medicine for treating traumatic fracture Download PDFInfo
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
- A61K36/704—Polygonum, e.g. knotweed
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D1/00—Surgical instruments for veterinary use
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/58—Reptiles
- A61K35/586—Turtles; Tortoises, e.g. terrapins
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/11—Pteridophyta or Filicophyta (ferns)
- A61K36/12—Filicopsida or Pteridopsida
- A61K36/126—Drynaria
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/232—Angelica
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/29—Berberidaceae (Barberry family), e.g. barberry, cohosh or mayapple
- A61K36/296—Epimedium
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- A—HUMAN NECESSITIES
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/46—Eucommiaceae (Eucommia family), e.g. hardy rubber tree
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/74—Rubiaceae (Madder family)
- A61K36/746—Morinda
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/80—Scrophulariaceae (Figwort family)
- A61K36/804—Rehmannia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/894—Dioscoreaceae (Yam family)
- A61K36/8945—Dioscorea, e.g. yam, Chinese yam or water yam
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
Abstract
The invention provides an application of a bone-protecting capsule as a medicine for treating traumatic fracture, and relates to the technical field of traditional Chinese medicine application. The invention relates to an application of a bone-protecting capsule which is prepared from traditional Chinese medicines including prepared fleece-flower root, epimedium herb, prepared rehmannia root, tortoise shell, morinda root, eucommia bark, teasel root, drynaria rhizome, Chinese angelica and Chinese yam as a medicine for treating traumatic fracture. The invention fully verifies that the bone-protecting capsule can promote bone formation, accelerate the generation of bony callus at the fracture part and improve the total healing efficiency of fracture by constructing a rat fracture model, thereby providing experimental basis for the medicine for treating traumatic fracture.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine application, in particular to application of a bone-protecting capsule as a medicine for treating traumatic fracture.
Background
Fractures disrupt the continuity and integrity of the bone. The problem that the slow healing of the fracture is the key problem of the invention is solved, western medicines for bone grafting, inflammation diminishing and bone formation promoting are mostly adopted in western medicines, and the western medicines have great side effects and high operation cost. The traditional Chinese medicine can improve the quality of callus and accelerate healing by improving the blood supply of the fracture part.
The traditional Chinese medicine has a long history of understanding and treating the fracture and has the advantages that western medicine and western medicine are difficult to compare. The Western medicine adopts operation fixation for treating severe fracture, and only antiphlogistic and analgesic medicines and no bone-knitting medicines are used after the operation. The manual reduction of the fracture and the small splint are matched with the traditional Chinese medicine for bone setting for oral administration or external application, which is a unique treatment scheme of the traditional Chinese medicine orthopedics and traumatology department and is widely applied to the treatment of the fracture. However, due to lack of systematic and intensive research and development, currently, most of the clinical use is plaster processed by an earth method, and commercially available bone-knitting Chinese patent medicines, such as fracture-setting tablets in traumatology, sanli powder and the like, are easy to accept by patients and use more because of convenient taking, but the drug effect is unclear, so that people hope to find more effective and safer novel bone-knitting Chinese patent medicines.
With the continuous and intensive research, the current international pathological research on fracture healing is intensive to the cellular and molecular level. The core event in fracture healing is bone remodeling: the absorption of calcified callus and the formation of new bone continue. It can be seen that in addition to new osteogenesis during fracture healing, bone resorption occurs simultaneously and both are in dynamic equilibrium, whereas the effector cells of both: how osteoblasts (Ob) and osteoclasts (Oc) maintain coupling balance between bone formation and bone resorption and regulation of molecules is not clear, and particularly, the mechanism of the treatment effect of the traditional Chinese medicine compound on the fracture is not researched much, so that the research on the fracture mechanism is a great hot spot at present.
The traditional Chinese medicine bone-protecting capsule is a new national patent drug produced by the exclusive company of Guangdong Annuo pharmaceutical industry GmbH and has the function of preventing and treating primary osteoporosis. The bone-protecting capsule is prepared from 10 traditional Chinese medicines of prepared fleece-flower root, prepared rehmannia root, tortoise shell, morinda officinalis, eucommia bark, teasel root, drynaria rhizome, Chinese angelica and Chinese yam, is prepared according to the principle of tonifying kidney and replenishing vital essence, strengthening spleen and replenishing qi, activating blood and dissolving stasis and relieving pain by applying the theory of traditional Chinese medicine, can strengthen tendons and bones, invigorate spleen and nourish blood, and activate blood and remove stasis, is clinically used for primary osteoporosis, and has obvious curative effect. Whether the traditional Chinese medicine bone-protecting capsule can be used as a traumatic fracture medicine or not has no relevant research at present.
The influence of the bone-protecting capsule on the bone structure and bone reconstruction state of a rat with ovariectomy is researched in the early stage, and the result shows that the bone-protecting capsule can increase the bone mass of a rat model with ovariectomy and osteoporosis and obviously promote bone formation, and the ALP content in serum is increased by promoting the proliferation and differentiation of osteoblasts and accelerating the absorption of calcium and phosphorus. The fracture healing is mainly related to bone formation, so that the effect of the bone-protecting capsule in the fracture healing is researched by applying a rat fracture model through methods such as imaging change of fracture ends, bone tissue morphometry and the like, and an experimental basis is provided for developing a new application of the bone-protecting capsule in treating traumatic fracture.
Disclosure of Invention
In view of the above, the invention establishes a rat fracture model, researches the effect of the bone-protecting capsule in fracture healing by methods such as imaging change of fracture ends, bone tissue morphometry and the like, and develops a new application of the bone-protecting capsule in treating traumatic fracture to provide experimental basis.
The invention aims to provide application of a bone-protecting capsule as a medicament for treating traumatic fracture, namely application of the bone-protecting capsule, which is prepared from prepared fleece flower root, epimedium herb, prepared rehmannia root, tortoise shell, morinda root, eucommia bark, teasel root, drynaria rhizome, Chinese angelica and Chinese yam serving as a traditional Chinese medicine formula, as the medicament for treating traumatic fracture.
Preferably, the bone-protecting capsule can promote bone formation, accelerate the generation of bony callus at the fracture part and improve the total healing efficiency of the fracture.
The invention also aims to provide a method for constructing a rat fracture model, which comprises the following steps:
after the rat is bred adaptively for 5 days, the model is made under the aseptic condition, 1cm periosteum is stripped in a sharp and annular mode at the middle section of the femur, the transverse fracture is caused by a osteotome, an 8-gauge needle head is penetrated in the patellar ligament of the knee joint in an antegrade mode, the transverse fracture is cut and buried in a marrow cavity, and then the rat fracture model is obtained by suturing layer by layer.
Preferably, according to the invention, the bone-protecting capsule is administered to the rat fracture model by gavage 1 time a day for 5 weeks; after the administration is finished, the effect of the bone-protecting capsule in traumatic fracture healing is researched by adopting X-ray to check fracture healing conditions, analyzing bone formation conditions of callus parts by bone morphometry and observing changes of bone fine structures of fracture parts by toluidine blue staining.
Compared with the prior art, the invention has the following beneficial effects:
the invention fully verifies that the bone-protecting capsule can promote bone formation, accelerate the generation of bony callus at the fracture part and improve the total healing efficiency of fracture by constructing a rat fracture model, thereby providing experimental basis for the medicine for treating traumatic fracture.
Drawings
FIG. 1 is a graph showing the change in body weight of rats in Ctrl group and HG group during administration;
FIG. 2 is a graph showing the results of the X-ray and gross examination of rats in Ctrl group and HG group after the administration;
FIG. 3 is a graph showing the effect of toluidine blue staining on the transverse section of the mid-femoral (callus) in rats in Ctrl and HG groups after the end of administration;
FIG. 4 is a graph showing the effect of toluidine blue staining on the cross section of mid-femoral (callus) in Ctrl group after the administration;
FIG. 5 is a graph showing the effect of fluorescence of calcein staining on the mid-femoral (callus) cross-sections of rats in Ctrl and HG groups after the end of administration.
Detailed Description
The present invention is further illustrated below with reference to specific experiments.
1 materials of the experiment
1.1 Experimental animals
SPF grade male and female SD rats, 3 months old, purchased from Guangdong province medical laboratory animal center, license number: SCXK (Yue) 2013-.
1.2 medicine and preparation method
Bone-protecting capsules (national Standard: Z20040124, production batch: 20170207, specification: 0.45 g/granule) provided by Guangdong Annuo pharmaceutical industry Co., Ltd; the bone-protecting capsule is prepared into a solution with the concentration of 0.12g/ml according to the conversion of the equivalent dose of the human body.
1.3 Primary reagents and instruments
Calcein (Sigma, lot # SLBRO830V), toluidine blue (Sigma, lot # 92-31-9); leica RM2500 type microtome, Leica SP1600 type saw; Bioquant-Osteo bone morphology image analysis system.
2 method of experiment
2.1 rat fracture model
After the rats are bred adaptively for 5 days, the rats are molded under an aseptic condition, 1cm periosteum is stripped in a sharp and annular mode at the middle section of the femur, a transverse fracture is caused by a osteotome, an 8-gauge needle head is penetrated in the patellar ligament of the knee joint in an antegrade mode, the rat is cut and buried in a marrow cavity, and then the rat is sutured layer by layer.
2.2 test grouping and administration
Rats were randomly divided into a control group (Ctrl) and a drug administration group (HG) by body weight, 6 each. After 2 days of molding, Ctrl and HG were perfused with stomach saline and HG, Qd, 4mL/kg/d, respectively, for 5 weeks. HG dose is converted into rat dose (0.486g/kg/d) and administration volume (0.12 g/ml) by standard animal equivalent dose conversion coefficient method, and is adjusted according to the change of rat body weight every week.
2.3 animal body weight
The dosing period rats were weighed 1 time per week as shown in figure 1.
2.4X-ray examination and scoring of fracture
After the administration, all rats were subjected to abdominal anesthesia and then X-ray radiographs were taken, and the scores were scored by X-ray examination using the fracture healing score criteria formulated in the treatise on traumatology, as shown in Table 1.
TABLE 1X-ray imaging scoring Table (mean + -s) after 5 weeks of dosing
P <0.05 compared to Ctrl set
2.5 bone microstructure at fracture site
After the administration is finished, the right femur is separated and taken down at the hip joint and fixed in 4% paraformaldehyde for 24-48 h. The fixed rat femur was embedded in non-decalcified hard tissue and sectioned at the callus for 5 μm, and the microstructure of the bone at the fracture site was observed by toluidine blue staining.
2.6 bone morphometry analysis
The living animal double-fluorescence labeling method is adopted. All rats were sacrificed by cardiac blood withdrawal at the end of the drug intervention. And (4) separating and taking down the right femur at the hip joint, and fixing the right femur in 4% paraformaldehyde for 24-48 h. The fixed rat femur was embedded in non-decalcified hard tissue and sectioned at the callus for 10 μm, bone morphometry analysis was performed to observe bone formation.
2.7 statistical methods
Statistical analysis was performed using GraphPad Prism 6.0. Data were analyzed by one-way anova, expressed as ± s, and P <0.05 indicated that the difference was statistically significant.
As can be seen in FIG. 2, the Ctrl group fracture lines are clear and bony callus is not seen on both sides of the fracture, but the broken end edges are still visible; and the fracture line of the HG group is basically disappeared, the fracture is basically healed, and part of the fracture is visible to be connected with the fracture end by bony callus.
As can be seen from FIGS. 3 and 4, the bifluorescence at callus of Ctrl group is less, indicating that osteoblasts are not highly active and bone formation is low. A large amount of bifluorescence exists at the callus of the HG group, which shows that the activity of osteoblasts is increased, the bone formation is increased, so that the fracture line of the rat of the HG group is basically disappeared, lamellar bones with regular distribution of bone trabeculae appear at the fracture end, and the bone connection is firmer. Meanwhile, a small amount of deep staining areas of callus in the Ctrl group are chondrocytes with larger volume, while chondrocytes are not found in callus in the HG group; in addition, in the dyeing result of the cross section of the callus at the fracture end, the callus matrix of the Ctrl group is mostly thick collagen fibers, the trabecular bone is immature, disorderly and loose in arrangement, i.e., mostly woven bone, while the protofracture of the HG group is basically bony callus, and regularly arranged trabecular bone appears after bone reconstruction.
Calcein causes the surface of the bone of the rat to generate green fluorescence, and as can be seen from figure 5, green fluorescence at the femoral callus of the Ctrl group is less and mostly single fluorescence; whereas HG group showed more green fluorescence and exhibited more bifluorescence at the femoral callus.
In conclusion, HG can promote bone formation, accelerate the generation of bony callus at the fracture part, improve the total healing efficiency of the fracture and provide experimental basis for the medicine for treating traumatic fracture.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (4)
1. The application of the bone-protecting capsule as a medicament for treating traumatic fracture is characterized in that the bone-protecting capsule which uses the traditional Chinese medicine formula of prepared fleece flower root, epimedium herb, prepared rehmannia root, tortoise shell, morinda root, eucommia bark, himalayan teasel root, fortune's drynaria rhizome, Chinese angelica and Chinese yam as the medicament for treating traumatic fracture.
2. The use of claim 1, wherein said bone-protecting capsule promotes bone formation, accelerates the formation of bony callus at the fracture site, and increases the overall efficiency of fracture healing.
3. The use according to claim 2, wherein a rat fracture model is first constructed and the bone-protecting capsule is administered to the rat fracture model by gavage, 1 time per day for 5 weeks; after the administration is finished, the effect of the bone-protecting capsule in traumatic fracture healing is researched by adopting X-ray to check fracture healing conditions, analyzing bone formation conditions of callus parts by bone morphometry and observing changes of bone fine structures of fracture parts by toluidine blue staining.
4. The use according to claim 3, wherein the rat fracture model is constructed by a method comprising the following steps:
after the rat is bred adaptively for 5 days, the model is made under the aseptic condition, 1cm periosteum is stripped in a sharp and annular mode at the middle section of the femur, the transverse fracture is caused by a osteotome, an 8-gauge needle head is penetrated in the patellar ligament of the knee joint in an antegrade mode, the transverse fracture is cut and buried in a marrow cavity, and then the rat fracture model is obtained by suturing layer by layer.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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2021
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