CN110840882A - Composition for treating osteoporosis - Google Patents
Composition for treating osteoporosis Download PDFInfo
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- CN110840882A CN110840882A CN201910855234.7A CN201910855234A CN110840882A CN 110840882 A CN110840882 A CN 110840882A CN 201910855234 A CN201910855234 A CN 201910855234A CN 110840882 A CN110840882 A CN 110840882A
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- bone
- composition
- osteoporosis
- isoimperatorin
- dihydromyricetin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
- A61K31/37—Coumarins, e.g. psoralen
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
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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
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Abstract
The invention discloses a composition for treating osteoporosis, which consists of the following raw materials in parts by weight: 2-10 parts of isoimperatorin and 1 part of dihydromyricetin. In addition, the invention also discloses application of the composition. The composition provided by the invention takes isoimperatorin as a main active ingredient, is matched with dihydromyricetin, can inhibit osteoclast differentiation and inhibit bone absorption, can promote osteoblast differentiation and bone formation, can play a role in treating osteoporosis by regulating balance of osteoblasts and osteoclasts, can obviously increase bone density of an osteoporosis model animal and restore bone microstructure, has obvious improvement on bone related parameters and increased bone formation amount, can effectively treat osteoporosis, and has the potential of being developed into a medicament, a health-care product or food for treating osteoporosis.
Description
Technical Field
The invention belongs to the technical field of medicines, health-care products and foods, and particularly relates to a composition for treating osteoporosis.
Background
Osteoporosis is a degenerative bone disease characterized by a decrease in bone mass and a deterioration in the microstructure of bone, resulting in increased bone fragility and susceptibility to fracture, and is a systemic metabolic bone disease. With the improvement of the average life span of human beings and the aggravation of the aging of population, senile osteoporosis becomes a global public health problem and seriously harms the physical health and the life quality of people. In developed countries in europe and america, tens of millions of people over 50 have osteoporosis or low bone mass, more than 400 million people with new osteoporosis and fracture every year, and the annual expenditure exceeds $ 500 million. China has already stepped into an aging society, and by the end of 2018, the latest data of the population of the aged people in China are as follows: 2.49 hundred million people in 60 years old and over, accounting for 17.9% of the total population; 1.67 hundred million people in 65 years and above, accounting for 11.9% of the total population; the aging of the population is accelerated, and the population over 60 years of age in China accounts for more than 20% of the population by 2025. The serious consequences of senile osteoporosis and fracture thereof, such as human mouth quality and the development of the economic society, are increasingly shown, the incidence rate of hip fracture in Beijing area is increased by nearly 3 times within 15 years, and the annual medical expense is 110 hundred million yuan. China starts a Chinese skeleton and health ten years action plan in 2002, and the prevention and treatment of senile osteoporosis becomes one of important basic problems in the field of health scientific research in China.
Osteoporosis is a complex pathogenesis in which osteoblasts and osteoclasts are in balance and one of the keys to maintaining normal bone function. The process of bone metabolism and bone turnover is the process by which osteoblasts form new bone and osteoclasts resorb old bone. Normal bone metabolism depends on a dynamic equilibrium between bone formation by osteoblasts and bone resorption by osteoclasts. If bone formation is greater than bone resorption, an increase in bone mass is manifested and vice versa as a decrease in bone mass, and severe bone mass reduction can lead to osteoporosis. The underlying mechanism by which osteoporosis occurs is an imbalance in the remodeling of body bone, i.e., the removal of old bone by osteoclasts (bone resorption) and the formation of new bone by osteoblasts (bone formation).
The existing western medicines have certain advantages in the aspect of controlling osteoporosis, play the role of resisting osteoporosis mainly by inhibiting bone resorption or promoting bone formation, but have single treatment target spot, narrow range, expensive medicine price and long medicine taking time, and have certain adverse drug reactions, so that many patients are difficult to accept. Therefore, the search for new drugs/drug combinations with low price, good safety and significant efficacy is the key to solving these problems.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a composition for treating osteoporosis, which has the advantages of obvious curative effect, low cost and small toxic and side effects, aiming at the defects of the prior art. The composition takes isoimperatorin as a main active ingredient, is matched with dihydromyricetin, can inhibit osteoclast differentiation and bone absorption, can promote osteoblast differentiation and bone formation, can play a role in treating osteoporosis by regulating balance of osteoblasts and osteoclasts, can obviously increase bone density of osteoporosis model animals, recover bone microstructure, remarkably improve bone related parameters (bone mineral density, bone body integral number, trabecular bone thickness, number and the like), increase bone formation amount, and can effectively treat osteoporosis.
In order to solve the technical problems, the invention adopts the technical scheme that: the composition for treating osteoporosis is characterized by comprising the following raw materials in parts by weight: 2-10 parts of isoimperatorin and 1 part of dihydromyricetin.
The composition for treating osteoporosis is characterized by comprising the following raw materials in parts by weight: 4-8 parts of isoimperatorin and 1 part of dihydromyricetin.
The composition for treating osteoporosis is characterized by comprising the following raw materials in parts by weight: 5 parts of isoimperatorin and 1 part of dihydromyricetin.
In addition, the invention also provides application of the composition in preparing a medicament for treating osteoporosis.
Furthermore, the invention also provides application of the composition in preparing health-care products for treating osteoporosis.
In addition, the invention also provides application of the composition in preparing food for treating osteoporosis.
The weight portion can be measured in units of weight such as gram, two, jin, kg and ton.
Compared with the prior art, the invention has the following advantages:
1. the composition of the invention takes isoimperatorin as a main active ingredient, is matched with dihydromyricetin, can inhibit osteoclast differentiation and inhibit bone resorption, can promote osteoblast differentiation and promote bone formation, can play a role in treating osteoporosis by regulating balance of osteoblasts and osteoclasts, can obviously increase bone density of osteoporosis model animals, recover bone microstructure, obviously improve bone related parameters (bone mineral density, bone body integral number, trabecular bone thickness, number and the like), and increase bone formation.
2. The raw materials of the composition are from plants, are cheap and easily available, have small toxic and side effects and high safety, can obviously improve osteoporosis symptoms, and is a pharmaceutical composition with the advantages of low price, safety, high efficiency and the like.
3. The composition can effectively treat osteoporosis, and has potential to be developed into a medicament, a health-care product or food for treating osteoporosis.
The technical solution of the present invention is further described in detail with reference to the accompanying drawings and embodiments.
Drawings
FIG. 1 is a graph showing the results of alkaline phosphatase staining after MC3T3-E1 cells treated with the composition of example 1 of the present invention;
FIG. 2 is a graph showing the results of alizarin red staining after MC3T3-E1 cells were treated with the composition of example 1 of the present invention;
FIG. 3 is a graph showing the results of detection of osteogenic differentiation marker genes ALP, OCN, Col I and Runx2 using RT-PCR detection after MC3T3-E1 cells were treated with the composition of example 1 of the present invention;
FIG. 4 is a TRAP staining of osteoclasts after induction by primary bone marrow mononuclear macrophages treated with the composition of example 1 of the present invention;
FIG. 5 is a graph showing the results of femoral bone density scans after administration of the composition of example 1 of the present invention to an osteoporotic mouse;
FIG. 6 is a micropt scan of a femur of an osteoporotic mouse after drenching with the composition of example 1 of the present invention;
FIG. 7 is a schematic diagram of the results of software quantitative analysis after femoral MicroCT scanning of FIG. 6;
FIG. 8 is a schematic diagram showing the histological examination results of femoral slices after administration of the composition of example 1 of the present invention to an osteoporosis mouse;
fig. 9 is a schematic diagram of the detection result of the liver and kidney function indexes after the composition of the embodiment 1 of the invention is infused into the osteoporosis mouse.
Detailed Description
Example 1
The composition for treating osteoporosis is prepared by uniformly mixing the following raw materials in parts by weight: 5 parts of isoimperatorin and 1 part of dihydromyricetin. The weight portion can be measured in units of weight such as gram, two, jin, kg and ton.
Example 2
The composition for treating osteoporosis is prepared by uniformly mixing the following raw materials in parts by weight: 2 parts of isoimperatorin and 1 part of dihydromyricetin. The weight portion can be measured in units of weight such as gram, two, jin, kg and ton.
Example 3
The composition for treating osteoporosis is prepared by uniformly mixing the following raw materials in parts by weight: 4 parts of isoimperatorin and 1 part of dihydromyricetin. The weight portion can be measured in units of weight such as gram, two, jin, kg and ton.
Example 4
The composition for treating osteoporosis is prepared by uniformly mixing the following raw materials in parts by weight: 10 parts of isoimperatorin and 1 part of dihydromyricetin. The weight portion can be measured in units of weight such as gram, two, jin, kg and ton.
Example 5
The composition for treating osteoporosis is prepared by uniformly mixing the following raw materials in parts by weight: 8 parts of isoimperatorin and 1 part of dihydromyricetin. The weight portion can be measured in units of weight such as gram, two, jin, kg and ton.
The composition of isoimperatorin and dihydromyricetin of example 1 of the present invention was tested as follows:
(1) effect of combination of Isoimperatorin and Dihydromyricetin on osteoblast differentiation
a. Culture of mouse preosteoblastic cell line MC3T3-E1 cells
MC3T3-E1 cells were cultured in complete medium (α -MEM solution supplemented with 10% fetal bovine serum, 10mM L-glutamine, 100. mu.g/mL penicillin, 100. mu.g/mL streptomycin sulfate), and the cells were incubated at 37 ℃ with 5% CO2Cultured in an incubator.
Osteogenic differentiation Induction of MC3T3-E1 cells
MC3T3-E1 preosteoblasts were cultured to a cell density of 80%, and MC3T3-E1 cells were subjected to osteogenic differentiation induction culture using an osteogenic differentiation medium (α -MEM solution supplemented with 10% fetal bovine serum, 10mM L-glutamine, 100U/mL penicillin, 100. mu.g/mL streptomycin sulfate, 0.005% vitamin C, and 10mM β -sodium glycerophosphate) as a control group, wherein 10% fetal bovine serum, 10mM L-glutamine, 100U/mL penicillin, 100. mu.g/mL streptomycin sulfate, 0.005% vitamin C, 10mM β -sodium glycerophosphate, isoimperatorin, and dihydromyricetin were added to the osteogenic differentiation medium of example 1 as a test group, and the final concentration of the composition of example 1 in the medium for two days was 20. mu.M, and the medium was changed once during induction culture, and was prepared as it was used.
c. Alkaline phosphatase (ALP) staining detection method
After 2 days of osteogenic differentiation induction culture of the cells, the level of osteogenic differentiation of the cells is detected by alkaline phosphatase staining, and the specific operation is as follows:
the medium was decanted and washed 3 times with PBS, 5min each; adding 0.5mL of 4% paraformaldehyde solution, and fixing at room temperature for 20 min; washing with PBS for 5min for 3 times; staining the cells with BCIP/NBT alkaline phosphatase staining kit (Shanghai Biyuntian Biotechnology Co., Ltd.), and incubating at 37 deg.C for 30 min; washing with deionized water for 5min for 2 times; drying at normal temperature, and scanning and storing by using a scanner.
Results referring to fig. 1, the depth of staining represents the differentiation level of osteoblasts. ALP staining was significantly increased after treatment with the isoimperatorin and dihydromyricetin composition (ZHW in the figure), indicating that the composition can enhance ALP activity and promote osteoblast differentiation.
d. Alizarin red staining detection method
After the cells are cultured for 14 days in osteogenic differentiation induction, the level of osteogenic differentiation of the cells is detected by alizarin red staining, and the specific operations are as follows:
the medium was decanted and washed 3 times with PBS, 5min each; adding 0.5mL of 4% paraformaldehyde solution, and fixing at room temperature for 20 min; washing with PBS for 5min for 3 times; staining the cells with 0.5% alizarin red solution (pH 4.2) and incubating at room temperature for 2 h; washing with distilled water for 5 times, each for 10 min; drying at normal temperature, scanning with scanner and storing.
Results referring to fig. 2, alizarin red staining detects calcified nodules in a medium deposited by extracellular matrix secreted by cells during osteoblast differentiation, and when alizarin red is used to stain the cells, the calcified nodules can be stained red by alizarin red, while extracellular matrix without calcified nodules is not stained after washing. Therefore, the amount of red nodules on the staining result can reflect the amount of calcium deposited during osteoblast differentiation, and the more calcium deposited, the higher the osteoblast differentiation degree. The number of red calcified nodules was significantly increased after treatment with the isoimperatorin and dihydromyricetin composition (ZHW in the figure), indicating that the composition can promote osteoblast differentiation.
e. Osteogenic differentiation related gene RT-PCR detection method
Two days after the MC3T3-E1 preosteoblasts are subjected to osteogenic differentiation induction culture, total RNA is extracted by using a Trizol method, and concentration detection is carried out by using a trace ultraviolet spectrophotometer. Reverse transcription was performed using One step PrimeScript RT reagent Kit and RT-PCR detection was performed using a Thermal Cycler C-1000 fluorescent quantitative PCR instrument, three duplicate wells were set for each set of sample experiments, and all loading was done on ice.
TABLE 1 reverse transcription reaction System
The reverse transcription procedure was: reverse transcription at 37 deg.C for 15min, and reverse transcriptase inactivation at 85 deg.C for 5 s.
The expression levels of Alp, Runx2 and Col-I, OCN in the induced MC3T3-E1 osteoblasts are detected.
The PCR primer sequences are shown in table 2:
TABLE 2 PCR primer sequences
Referring to FIG. 3, after treatment with the combination of isoimperatorin and dihydromyricetin, the expression levels of the osteogenic differentiation marker genes ALP, OCN, Col I and Runx2 in MC3T3-E1 cells were increased compared to the control group (.: p < 0.01;. p < 0.001). It is demonstrated that the above composition can promote osteoblast differentiation.
(2) Effect of the combination of Isoimperatorin and Dihydromyricetin on osteoclast differentiation
a. Mouse primary bone marrow mononuclear macrophage (BMMs) extraction
Killing a 6-week-old mouse after removing the neck, placing the mouse in a beaker containing 75% ethanol, and soaking and disinfecting the whole body for 3 min; placing the mouse in
The skin and muscle of the mouse were carefully removed using sterile surgical scissors and forceps, the femur and tibia were packed and separated using sterile gauze and placed in a sterile petri dish, it was noted that, from the removal of skin and muscle to the separation of femur and tibia, the bone tissue was not accessible during the procedure, the scissors and forceps for dissecting the outer skin were used separately from the scissors and forceps for stripping the muscle and placed back in 75% ethanol in time for sterilization, and the separated femur and tibia were placed in 5mL of α -MEM medium containing diabodies
Taking out bone tissue, inoculating cells into a culture dish for culture, wherein the whole process cannot be more than 2h, removing epiphyses at the tail end of a marrow cavity by using a sterile surgical scissors, using a 5mL syringe together with a 1mL syringe needle to insert into the marrow cavity, sucking α -MEM culture medium containing double antibodies to flush out bone marrow to a 15mL sterile centrifuge tube, repeatedly flushing until the femur and the tibia become ivory, filtering flushed out bone marrow culture solution into the sterile 15mL centrifuge tube by using a 70 mu m cell filter screen, centrifuging at 800rpm for 6min, discarding supernatant, adding 2mL erythrocyte lysate into a bone marrow cell sediment extracted from each mouse, gently blowing the erythrocyte lysate to about 20 times by using a pipette, slowly mixing the erythrocyte lysate at room temperature for 5min, centrifuging at 800rpm for 6min, removing erythrocytes in the marrow cells, discarding supernatant, adding 1mL complete culture medium (α -MEM + 10% FBS + double antibody suspension) into the bone marrow cell sediment, and re-suspending the cells
The sterile culture dish of (1) was added with M-CSF to a final concentration of 10ng/mL, and the mixture was placed at 37 ℃ and 5% CO2Culturing in an incubator overnight; the next day, the suspension cells in the culture medium are collected in a 15mL centrifuge tube and centrifuged at 800rpm for 6 min; primary BMMs cells were obtained.
b. In vitro osteoclast differentiation induction by primary bone marrow mononuclear macrophage
Osteoclast differentiation was induced in vitro in BMMs cells using an osteoclast induction medium (complete medium supplemented with 10ng/mL M-CSF and 10ng/mL RANKL). The specific induction method is as follows: primary BMMs cells extracted from mouse bone marrow at 1X 105Osteoclast induction can be performed by inoculating cells/well in 48-well plates and observing that the adherent BMMs account for more than 20% of the total amount of the cells. The original culture medium supernatant was discarded, and the medium was replaced with an osteoclast induction medium to start induction. And continuously supplementing the osteoclast induction culture medium into the holes on the next day without discarding the culture medium, discarding the culture medium on the third day, replacing the fresh osteoclast induction culture medium, and continuously supplementing the osteoclast induction culture medium on the fourth day, namely circularly replacing the fresh osteoclast induction culture medium every two days. The experiment was divided into a control group in which the primary BMMs were normally induced by adding an osteoclast induction medium to the primary BMMs, and an administration group in which the primary BMMs were simultaneously added with an osteoclast induction medium and 5. mu.M of a composition of isoimperatorin and dihydromyricetin of example 1 of the present invention. Mature multinuclear osteoclast can be observed in the control group about 4 days after the osteoclast induction culture, and the sample collection and detection can be carried out.
c. Osteoclast TRAP staining
Mouse femoral bone tissue sections were TRAP stained using the Acid Phosphatase Leukocyte (TRAP) Kit manufactured by Sigma-Aldrich, USA.
The preparation method of the TRAP staining solution is as follows:
preparing sufficient deionized water, and heating in a water bath at 37 ℃; adding 100 mu LFast garnet GBC base Solution and 100 mu L Sodium nitrate Solution into a 1.5mL sterile centrifuge tube, gently mixing for 30s, and standing for 2min at room temperature; taking 10mL of a sterile centrifuge tube, and preparing TRAP staining solution according to the table 3;
TABLE 3 TRAP staining solution composition and dosage
Osteoclast TRAP staining
After the primary BMMs cells are cultured for 4 days by inducing an osteoclast induction medium, TRAP staining is carried out on mature osteoclasts. The method comprises the following specific steps:
discarding the culture medium, and gently rinsing with PBS (phosphate buffer solution) for 1-2 times, 3-5 min each time; adding 4% paraformaldehyde solution, and fixing for 20 min; discarding the stationary liquid, rinsing with PBS buffer solution for 2-3 times, 3-5 min each time; discarding PBS buffer solution, adding appropriate amount of TRAP staining solution (adding 200 μ L per well of 48-well plate), and incubating at 37 deg.C in dark for 1 h; after the incubation is finished, discarding the staining solution, and rinsing for 3 times by using PBS buffer solution to terminate the reaction; the results of cell staining were observed using an optical microscope and photographed.
Referring to fig. 4, after the combined action of isoimperatorin and dihydromyricetin, RANKL-induced TRAP enzyme activity was inhibited, staining positive cells were decreased, and the number of mature multinucleated osteoclasts was decreased, indicating that the combination of isoimperatorin and dihydromyricetin can inhibit osteoclast differentiation.
(3) Therapeutic effect of combination of isoimperatorin and dihydromyricetin on osteoporosis mice
A15-month-old C57BL/6 mouse was selected as an animal model for osteoporosis, and a 3-month-old C57BL/6 mouse was selected as a control, and the therapeutic effect of the composition of isoimperatorin and dihydromyricetin of example 1 of the present invention on osteoporosis was examined. The combination of isoimperatorin and dihydromyricetin was administered to mice by gavage once daily for 7 weeks at a dose of 50 mg/kg.
a. Bone density detection in mice
After anesthetizing the mice with sodium pentobarbital, the bone density of the mice after administration was measured using a high resolution dual energy X-ray imaging system (MEDIKORS, InAlyzer, 55KeV/80 KeV).
Referring to fig. 5, the femoral bone density of 15-month-old osteoporosis mice was significantly lower than that of young control mice; after the composition of isoimperatorin and dihydromyricetin is irrigated, the femoral bone density of an osteoporosis mouse is obviously increased, which shows that the composition can improve osteoporosis symptoms.
b. Micro CT detection of femur morphology of mouse
The mouse is killed by taking off the neck, the femur is taken out by stripping, the soft tissue on the surface of the bone is removed, after the fixation with 70% ethanol, a micro CT scanner is adopted for scanning, and after data is obtained, the following parameters are analyzed by special bone analysis software: bone density (BMD), bone volume fraction (BV/TV%), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular resolution (Tb.Sp), etc.
Referring to fig. 6 and 7, the bone density, the bone volume fraction, the trabecular bone thickness and the trabecular bone number of 15-month-old osteoporosis mice were all significantly reduced, and the trabecular bone separation was increased; after administration of isoimperatorin and dihydromyricetin combination, there was a significant recovery in bone density, bone mass fraction, trabecular thickness, trabecular number and trabecular separation ([ p ] 0.05; [ p ] 0.01; [ p ] 0.001), indicating that aging decreased femoral bone density and bone microstructure in mice, whereas treatment with isoimperatorin and dihydromyricetin combination increased femoral bone density and bone microstructure in mice. The composition of isoimperatorin and dihydromyricetin can cure osteoporosis.
c. Mouse femoral histology detection
The mice were sacrificed by cervical dislocation, the femurs were removed and fixed with 4% paraformaldehyde, and then decalcified with 14% EDTA solution for 28 days. Paraffin sections were stained with H & E and bone tissue morphology was observed under a microscope.
Referring to fig. 8, femoral sections show a significant reduction in trabecular bone number in 15-month-old osteoporotic mice compared to 3-month-old control mice. The bone is sparsely fractured, most of the bone cannot be connected into a net, a large blank area appears in a bone trabecular structure, and a large amount of fat cells are visible. After the treatment of the isoimperatorin and dihydromyricetin composition (ZHW in the figure), the trabecular bone of the femur is obviously wide and thick, the number of the trabecular bone is also obviously increased, and the fracture of the trabecular bone and the occurrence of fat cells are rare.
(4) Detection of liver and kidney toxicity of mouse by using isoimperatorin and dihydromyricetin composition
Blood was drawn from the mice after the administration by cardiac puncture, serum was separated, and ALT, AST, r-GT and BUN levels in the serum were measured.
Referring to FIG. 9, there was no significant change in the mean levels of liver and kidney function indices ALT, AST, r-GT, and BUN in mice treated with the isoimperatorin and dihydromyricetin composition compared to the control group. It is demonstrated that the application of the above composition for a prolonged period of time (7 weeks) has no significant hepatorenal toxicity and is safer.
The above tests were carried out on the compositions of isoimperatorin and dihydromyricetin of examples 2 to 5 of the present invention, and the results were similar.
In conclusion, the composition has the advantages of remarkable curative effect, low cost and small toxic and side effects, can be used for treating osteoporosis, and can be used for preparing medicines, health-care products and foods for treating osteoporosis.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, changes and equivalent structural changes made to the above embodiment according to the technical spirit of the present invention still fall within the protection scope of the technical solution of the present invention.
Claims (6)
1. The composition for treating osteoporosis is characterized by comprising the following raw materials in parts by weight: 2-10 parts of isoimperatorin and 1 part of dihydromyricetin.
2. The composition for treating osteoporosis of claim 1, which is prepared from the following raw materials in parts by weight: 4-8 parts of isoimperatorin and 1 part of dihydromyricetin.
3. The composition for treating osteoporosis of claim 2, which is prepared from the following raw materials in parts by weight: 5 parts of isoimperatorin and 1 part of dihydromyricetin.
4. Use of a composition according to any one of claims 1 to 3 in the manufacture of a medicament for the treatment of osteoporosis.
5. Use of a composition according to any one of claims 1 to 3 for the preparation of a health product for the treatment of osteoporosis.
6. Use of a composition according to any one of claims 1 to 3 for the preparation of a food product for the treatment of osteoporosis.
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| CN113648306A (en) * | 2021-08-16 | 2021-11-16 | 西北工业大学 | Application of bergamottin in preventing or treating osteoporosis and/or bone loss |
| CN115671096A (en) * | 2022-11-07 | 2023-02-03 | 西北工业大学 | Application of 6-methoxy angelicin and structural analogue thereof |
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| LIBO ZHAO等: "Dihydromyricetin Protects against Bone Loss in Ovariectomized Mice by Suppressing Osteoclast Activity", 《FRONTIERS IN PHARMACOLOGY》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113648306A (en) * | 2021-08-16 | 2021-11-16 | 西北工业大学 | Application of bergamottin in preventing or treating osteoporosis and/or bone loss |
| CN115671096A (en) * | 2022-11-07 | 2023-02-03 | 西北工业大学 | Application of 6-methoxy angelicin and structural analogue thereof |
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