CN111773265A - Application of dalbergia odorifera leaf extract in preparation of medicines for preventing or treating osteoporosis - Google Patents
Application of dalbergia odorifera leaf extract in preparation of medicines for preventing or treating osteoporosis Download PDFInfo
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- CN111773265A CN111773265A CN202010803663.2A CN202010803663A CN111773265A CN 111773265 A CN111773265 A CN 111773265A CN 202010803663 A CN202010803663 A CN 202010803663A CN 111773265 A CN111773265 A CN 111773265A
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- 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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Abstract
The invention provides an application of a dalbergia odorifera leaf extract in preparation of a medicine for preventing or treating osteoporosis. The dalbergia odorifera leaf extract can be obtained by the following method: extracting chemical components of dalbergia odorifera leaves with water, ethanol or methanol solvent, filtering the extracting solution to obtain filtrate, and concentrating the filtrate under reduced pressure to obtain the dalbergia odorifera leaf extract. Pharmacological test researches of the invention show that the dalbergia odorifera leaf extract has good anti-osteoporosis effect and can be used for preparing medicines for preventing or treating osteoporosis.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to application of a dalbergia odorifera leaf extract in preparation of a medicine for preventing or treating osteoporosis.
Background
Osteoporosis is a systemic metabolic bone disease with the clinical manifestations of decreased bone density, destruction of bone microarchitecture, increased bone fragility and frequent fractures. In 2010, world health organization statistics have shown that there are more than 2 billion osteoporosis patients worldwide, with one third of postmenopausal women. The rapid decline in estrogen levels in the body, leading to increased bone metabolism disorders, is a major cause of susceptibility to osteoporosis in postmenopausal women. The traditional estrogen replacement therapy can cause the patients to easily generate side effects such as high blood coagulation, hypertension, edema and the like, and greatly increase the risk of malignant tumors such as endometrial cancer, breast cancer and the like. In addition, the adoption of western medicines such as bisphosphate, calcitonin and fluoride to treat postmenopausal osteoporosis can also cause serious side effects including irritative esophagitis and generation of antigen reaction. Therefore, the search for safer and more reliable drugs for treating postmenopausal osteoporosis is the focus of long-term attention in the field of medicine research in the world.
The phytoestrogen is a non-steroidal compound with estrogen-like effect, the biological activity of the phytoestrogen is 1/500-1/1000 of estrogen, the phytoestrogen has mild effect and small toxic and side effect, and the phytoestrogen has wide application prospect in treating osteoporosis. Early literature suggests that: dalbergia odorifera (f) Dalbergia odoriferaDalbergia odorifera) The Hainan Huanghua pear is an important source of non-steroidal phytoestrogen and is also a basic source plant of rare medicinal materials dalbergia wood. However, Dalbergia odorifera leaves, especially deciduous leaves, have not been developed and utilized. The study of the subject group in the earlier stage finds that: the dalbergia odorifera leaf is rich in flavonoid components, and the extract of the dalbergia odorifera leaf has good anti-osteoporosis activity. The invention can provide reference for developing safe and effective anti-osteoporosis medicines in the future.
Disclosure of Invention
The invention provides the application of the dalbergia odorifera leaf extract in preparing the medicine for preventing or treating osteoporosis, overcomes the defects of the prior art, solves the problem that the dalbergia odorifera leaf, especially the dalbergia odorifera leaf, is not developed and utilized, improves the utilization rate of the dalbergia odorifera leaf resource, and especially relates to the application of the dalbergia odorifera leaf extract in preparing the medicine for preventing and treating osteoporosis.
One of the technical schemes of the invention is realized by the following modes: application of lignum Dalbergiae Odoriferae leaf extract in preparing medicine for preventing or treating osteoporosis is provided.
The second technical scheme of the invention is realized by the following mode: the preparation method of the dalbergia odorifera leaf extract comprises the following steps: extracting Dalbergia odorifera leaf with water, ethanol or methanol to obtain filtrate, and concentrating under reduced pressure to obtain Dalbergia odorifera leaf extract; wherein, water extraction: adding 1-10 times of water into Dalbergia odorifera leaf, extracting at 60-100 deg.C for 1-3 times (each for 1-3 hr), filtering, and mixing filtrates; ethanol extraction: adding 1-10 times of 20-90% ethanol into dalbergia odorifera leaves, extracting at 60-100 deg.C for 1-3 times (each for 1-3 hr), filtering, and mixing filtrates; methanol extraction: adding 1-10 times of methanol into dalbergia odorifera leaves, performing ultrasonic extraction for 1-3 hours to obtain an extracting solution, and filtering to obtain a filtrate.
The dalbergia odorifera leaf extract has a good anti-osteoporosis effect, can be further applied to medicines for preventing or treating osteoporosis, solves the problem that dalbergia odorifera leaves, particularly dalbergia odorifera fallen leaves are not developed and utilized, and improves the utilization rate of dalbergia odorifera leaf resources.
Detailed Description
The present invention is not limited by the following examples, and specific embodiments can be determined according to the technical solutions and practical situations of the present invention described above.
The invention is further discussed below with reference to examples. The use of the dalbergia odorifera leaf extract for preparing a medicament for preventing or treating osteoporosis of the present invention is explained in detail by the following examples:
example 1
The dalbergia odorifera leaf extract is applied to the preparation of medicines for preventing or treating osteoporosis, and is obtained by the following method:
the dalbergia odorifera leaf extract is prepared by using water as an extraction solvent, performing warm-immersion extraction at 100 ℃ according to the material-liquid ratio of 1:10 for 3 times and 2 hours each time, filtering, combining filtrates, and concentrating under reduced pressure.
Example 2
The dalbergia odorifera leaf extract is applied to the preparation of medicines for preventing or treating osteoporosis, and is obtained by the following method:
the dalbergia odorifera leaf extract is prepared by using 70% ethanol as an extraction solvent, performing warm-immersion extraction at 80 ℃ according to the material-liquid ratio of 1:5 for 3 times, filtering each time for 2 hours, combining filtrates, and concentrating under reduced pressure.
Example 3
The dalbergia odorifera leaf extract is applied to the preparation of medicines for preventing or treating osteoporosis, and is obtained by the following method:
carrying out ultrasonic extraction on dalbergia odorifera leaves for 2 hours according to the material-liquid ratio of 1:8 by taking methanol as an extraction solvent to obtain an extracting solution, filtering out a filtrate, and carrying out reduced pressure concentration to obtain the dalbergia odorifera leaf extract.
Example 4
The dalbergia odorifera leaf extract is applied to the preparation of medicines for preventing or treating osteoporosis, and is obtained by the following method:
the dalbergia odorifera leaf extract is prepared by using water as an extraction solvent, performing warm-immersion extraction at 60 ℃ according to the material-liquid ratio of 1:1 for 1 time and 3 hours each time, filtering out a filtrate, and performing reduced pressure concentration.
Example 5
The dalbergia odorifera leaf extract is applied to the preparation of medicines for preventing or treating osteoporosis, and is obtained by the following method:
the dalbergia odorifera leaf extract is prepared by using water as an extraction solvent, performing warm-immersion extraction at 80 ℃ according to a material-liquid ratio of 1:6 for 2 times, each time for 1 hour, filtering, combining filtrates, and concentrating under reduced pressure.
Example 6
The dalbergia odorifera leaf extract is applied to the preparation of medicines for preventing or treating osteoporosis, and is obtained by the following method:
the dalbergia odorifera leaf extract is prepared by using 20% ethanol as an extraction solvent, performing warm-immersion extraction at 60 ℃ according to the material-liquid ratio of 1:2 for 2 times, each time for 1 hour, filtering, combining filtrates, and concentrating under reduced pressure.
Example 7
The dalbergia odorifera leaf extract is applied to the preparation of medicines for preventing or treating osteoporosis, and is obtained by the following method:
the dalbergia odorifera leaf extract is prepared by using 90% ethanol as an extraction solvent, performing warm-immersion extraction at 100 ℃ according to the material-liquid ratio of 1:10 for 1 time and 3 hours each time, filtering out filtrate, and concentrating under reduced pressure.
Example 8
The dalbergia odorifera leaf extract is applied to the preparation of medicines for preventing or treating osteoporosis, and is obtained by the following method:
carrying out ultrasonic extraction on dalbergia odorifera leaves for 1 hour according to the material-liquid ratio of 1:3 by using methanol as an extraction solvent to obtain an extracting solution, filtering out a filtrate, and carrying out reduced pressure concentration to obtain the dalbergia odorifera leaf extract.
Example 9
The dalbergia odorifera leaf extract is applied to the preparation of medicines for preventing or treating osteoporosis, and is obtained by the following method:
carrying out ultrasonic extraction on dalbergia odorifera leaves by taking methanol as an extraction solvent according to the material-liquid ratio of 1:10 for 3 hours to obtain an extracting solution, filtering out a filtrate, and carrying out reduced pressure concentration to obtain the dalbergia odorifera leaf extract.
The following specific pharmacological tests of the use of the dalbergia odorifera leaf extract in the preparation of a medicament for preventing or treating osteoporosis according to the above example:
1. MTT method for measuring cell proliferation rate of mouse osteoblast MC3T3-E1 (subclone 14):
(1) the MC3T3-E1 cells in the dish were placed at 37 ℃ in 5% CO2The incubator is used for culture. Cell cultureThe nutrient solution comprises a DMEM culture medium, 10% heat-inactivated Fetal Bovine Serum (FBS), 100U/mL penicillin and 100 mug/mL streptomycin, and the nutrient solution is changed for 1 time in 2-3 days and is passaged for 1 time in 3-4 days.
(2) Collecting MC3T3-E1 cell 2 × 104Each well was inoculated into a 96-well plate to perform cell proliferation assay, and 200. mu.L of culture medium was placed in each well. The experimental group was added with dalbergia odorifera leaf extract at different final concentrations, the control group was added with DMEM of equal volume, and each group had 6 duplicate wells. The incubation was continued for 24, 48, 72, 96 h, 4h before the end of the incubation, 5 mg/mL (0.5%) of MTT 20. mu.L was added to each well. And (3) continuously culturing for 4h in the incubator, taking out, absorbing and discarding the culture solution, rinsing with PBS once, adding 150 mu L of DMSO into each hole, shaking and dissolving for 10-30 min by Minisaker, incubating for 15 min at room temperature in a dark place, reading the light absorption value of each hole at the wavelength of 570 nm by using an enzyme labeling instrument after crystals are fully dissolved, and calculating the cell proliferation rate.
2. Alkaline phosphatase (ALP) activity assay:
allowing lignum Dalbergiae Odoriferae leaf extract with different concentrations to act on cells for 24, 48, 72, and 96 h, discarding cell culture solution, gently washing with PBS for 2 times, treating with 0.2% TritonX-100 lysate 100 μ L, and lysing cells on ice for 30min, and collecting the treated sample at 4 deg.C and 12000 ×gCentrifuging for 15 min under the centrifugal condition. ALP values and protein contents were measured by ALP detection kit and BCA method, respectively, and intracellular ALP activity was calculated and expressed as U/(g. port). The supernatant was taken to determine ALP activity and protein concentration. Adding 1.0 mg/mL protein supernatant into 100 μ L−1pNPP、1mol·L−1Diethanolamine and 0.5 mmol. L−1MgCl2The buffer solution (2) is 100 mu L, incubated for 30min at 37 ℃ in the dark, and 3 mol. L is added into each well−1The reaction was stopped with 50. mu.L NaOH and the light absorption was measured at a wavelength of 405 nm. The BCA method measures the sample protein concentration for correcting ALP activity.
Through carrying out research on bone formation promoting activity on the dalbergia odorifera leaf extract, as shown in table 1, the results show that: compared with a control group, the Dalbergia odorifera leaf extract can significantly improve the OD value under different administration concentrations; the leaf extract of Dalbergia odorifera was found to be at 10 deg.C compared to the control group-6、10-7And 10-8Concentration of drug administrationNext, ALP activity can be significantly improved. Therefore, the dalbergia odorifera leaf extract has strong bone formation promoting activity.
TABLE 1 Dalbergia odorifera leaf extract on osteoblast Activity
Note: compared with the control groupP<0.05，**P<0.01。
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.
Claims (2)
1. An application of Dalbergia odorifera leaf extract in preparing medicine for preventing or treating osteoporosis is provided.
2. The use as claimed in claim 1, wherein the preparation method of the dalbergia odorifera leaf extract comprises: extracting Dalbergia odorifera leaf with water, ethanol or methanol to obtain filtrate, and concentrating under reduced pressure to obtain Dalbergia odorifera leaf extract; wherein, water extraction: adding 1-10 times of water into Dalbergia odorifera leaf, extracting at 60-100 deg.C for 1-3 times (each for 1-3 hr), filtering, and mixing filtrates; ethanol extraction: adding 1-10 times of 20-90% ethanol into dalbergia odorifera leaves, extracting at 60-100 deg.C for 1-3 times (each for 1-3 hr), filtering, and mixing filtrates; methanol extraction: adding 1-10 times of methanol into dalbergia odorifera leaves, performing ultrasonic extraction for 1-3 hours to obtain an extracting solution, and filtering to obtain a filtrate.
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| CN202010803663.2A CN111773265A (en) | 2020-08-11 | 2020-08-11 | Application of dalbergia odorifera leaf extract in preparation of medicines for preventing or treating osteoporosis |
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Citations (6)
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|---|---|---|---|---|
| CN102119957A (en) * | 2011-03-11 | 2011-07-13 | 浙江大学 | Preparation method and application of effective ingredient of rosewood heart wood |
| CN102204949A (en) * | 2011-05-09 | 2011-10-05 | 江南大学 | Method for preparing dalbergia odorifera seed extract and quickly screening anticoagulant activity of dalbergia odorifera seed extract |
| CN103773820A (en) * | 2012-10-22 | 2014-05-07 | 东北林业大学 | Method for extracting, separating and purifying isoflavone active components biochanin A and genistein from dalbergia odorifera T.Chen leaves |
| CN110776409A (en) * | 2019-11-07 | 2020-02-11 | 岳阳市中医医院 | Method for extracting pterocarpus indicus and application of extract in antitumor drugs |
| CN110845319A (en) * | 2019-11-28 | 2020-02-28 | 江西中医药大学 | Preparation method of neoflavonoid compound and application of neoflavonoid compound in preparation of medicine for treating osteoporosis |
| CN112755063A (en) * | 2021-01-19 | 2021-05-07 | 江西中医药大学 | Application of dalbergia odorifera leaf extract in preparation of hypolipidemic drugs and effervescent tablets thereof |
-
2020
- 2020-08-11 CN CN202010803663.2A patent/CN111773265A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102119957A (en) * | 2011-03-11 | 2011-07-13 | 浙江大学 | Preparation method and application of effective ingredient of rosewood heart wood |
| CN102204949A (en) * | 2011-05-09 | 2011-10-05 | 江南大学 | Method for preparing dalbergia odorifera seed extract and quickly screening anticoagulant activity of dalbergia odorifera seed extract |
| CN103773820A (en) * | 2012-10-22 | 2014-05-07 | 东北林业大学 | Method for extracting, separating and purifying isoflavone active components biochanin A and genistein from dalbergia odorifera T.Chen leaves |
| CN110776409A (en) * | 2019-11-07 | 2020-02-11 | 岳阳市中医医院 | Method for extracting pterocarpus indicus and application of extract in antitumor drugs |
| CN110845319A (en) * | 2019-11-28 | 2020-02-28 | 江西中医药大学 | Preparation method of neoflavonoid compound and application of neoflavonoid compound in preparation of medicine for treating osteoporosis |
| CN112755063A (en) * | 2021-01-19 | 2021-05-07 | 江西中医药大学 | Application of dalbergia odorifera leaf extract in preparation of hypolipidemic drugs and effervescent tablets thereof |
Non-Patent Citations (1)
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| 范竹鸣，等: ""降香化学成分和药理作用研究进展"", 《时珍国医国药》 * |
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