CN111686147A - Eucommia ulmoides extract and application thereof in treating osteoporosis - Google Patents
Eucommia ulmoides extract and application thereof in treating osteoporosis Download PDFInfo
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- CN111686147A CN111686147A CN202010598494.3A CN202010598494A CN111686147A CN 111686147 A CN111686147 A CN 111686147A CN 202010598494 A CN202010598494 A CN 202010598494A CN 111686147 A CN111686147 A CN 111686147A
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- 241000208689 Eucommia ulmoides Species 0.000 title claims abstract description 82
- 239000000284 extract Substances 0.000 title claims abstract description 50
- 208000001132 Osteoporosis Diseases 0.000 title claims abstract description 22
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Classifications
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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)
- A61K36/46—Eucommiaceae (Eucommia family), e.g. hardy rubber tree
-
- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- Orthopedic Medicine & Surgery (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
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- Bioinformatics & Cheminformatics (AREA)
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Abstract
The invention provides an eucommia ulmoides extract and application thereof in treating osteoporosis, and belongs to the technical field of traditional Chinese medicine extraction. The eucommia ulmoides extract comprises the following components in parts by weight: 20-50 parts of eucommia ulmoides petroleum ether extract and 10-30 parts of eucommia ulmoides ethanol extract, wherein the weight ratio of the eucommia ulmoides petroleum ether extract to the eucommia ulmoides ethanol extract is controlled to be 1-5: the invention also provides an extraction method of the eucommia petroleum ether extract and the eucommia ethanol extract, because different active ingredients have different solubilities in different solvents, the content of the active ingredients can be effectively adjusted by adopting different solvents for extraction, the extraction and separation method is simple and easy to implement, and the effect of treating osteoporosis is good.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine extraction, and particularly relates to an eucommia ulmoides extract and application thereof in treating osteoporosis.
Background
Osteoporosis is a systemic skeletal disease characterized by decreased bone mass, destruction of the microstructure of bone tissue, increased bone fragility and increased fracture risk, and its clinical symptoms are mostly bone pain and fracture, which is a chronic disease caused by multiple factors and has a long treatment period. Although the drugs clinically used for treating osteoporosis at present, such as calcitonin, bisphosphonates, parathyroid hormone, estrogen, selective estrogen receptor modulators and the like, have good curative effects, the drugs used for a long time have negative effects, and researches show that the existing drugs can only thicken thinned trabeculae but can not connect broken trabeculae, so that the broken bone tissue microstructures can not be completely repaired.
The traditional Chinese medicine therapy is a traditional treatment method in China, the traditional Chinese medicine for treating the osteoporosis emphasizes the integral prevention and treatment, has small side effect, and has unique advantage on treating the chronic disease of the osteoporosis. The traditional Chinese medicine considers that the occurrence of osteoporosis is mainly related to three factors of kidney deficiency, spleen deficiency and blood nodule, wherein the kidney deficiency is the main cause of the disease.
Eucommia ulmoides (Eucommia ulmoides Oliv.) is a perennial deciduous tree belonging to Eucommia of Eucommiaceae, is a unique medicinal plant in China, is a rare nourishing medicinal material, has sweet and slightly pungent taste and warm property, and enters liver and kidney meridians. Eucommia ulmoides has been used as a medicine for more than 2000 years. It is listed as the top grade in Shen nong Bai Cao Jing (Shen nong's herbal Jing), and is indicated for lumbago, tonify middle energizer, replenish vital essence and energy, strengthen tendons and bones, remove yin, relieve itching and dampness, and dribbling urine. It is suitable for long-term administration, and has effects of reducing weight and resisting aging. The functions and indications of eucommia ulmoides recorded in the pharmacopoeia of the people's republic of China (2010 edition) are as follows: tonify liver and kidney, strengthen tendons and bones, prevent abortion. Can be used for treating deficiency of liver and kidney, soreness of waist and knees, weakness of tendons and bones, dizziness, blood drip during pregnancy, and threatened abortion. Eucommia ulmoides is widely used as a traditional good medicine for tonifying kidney and treating osteoporosis.
For example, chinese patent application 201310027831.3 discloses an extract of eucommia ulmoides oliv, which is prepared by the following steps: (1) extracting eucommia ulmoides serving as an extraction raw material with a mixed solution of alcohol and water; (2) removing pigment from the extracting solution in the step (1) through a neutral alumina chromatographic column, collecting eluent and concentrating; (3) the eucommia ulmoides extract is obtained by extracting the eluent obtained in the step (2) by using an organic solvent, and the application of the eucommia ulmoides extract in preparing a pharmaceutical composition for preventing and treating bone metabolism disorder diseases is provided, but the treatment effect is not ideal.
In order to achieve better treatment effect, in chinese patent application 201210250590.4, a composition comprising a combination of the traditional Chinese medicinal materials teasel root and eucommia bark, including a mixture of extracts obtained by separately extracting teasel root and eucommia bark, or an extract obtained by jointly extracting teasel root and eucommia bark, is selected from eucommia bark. The composition has good effect of treating osteoporosis, but the content of effective components in the mixed extract is relatively complex, and the content of various effective components is not easy to control.
Therefore, there is a need to develop an eucommia ulmoides extract with good and fast curative effect for better and more fundamentally treating osteoporosis.
Disclosure of Invention
The invention aims to provide an eucommia ulmoides extract and application of the eucommia ulmoides extract in preparing a pharmaceutical composition for treating and preventing osteoporosis.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
in one aspect, the invention provides an eucommia ulmoides extract, which comprises an eucommia ulmoides petroleum ether extract and an eucommia ulmoides ethanol extract.
The eucommia ulmoides extract comprises the following components in parts by weight: 20-50 parts of eucommia petroleum ether extract and 10-30 parts of eucommia ethanol extract.
Preferably, the eucommia ulmoides extract comprises the following components in parts by weight: 25-40 parts of eucommia petroleum ether extract and 13-25 parts of eucommia ethanol extract.
Still preferably, the eucommia ulmoides extract comprises, by weight: 30-48 parts of eucommia petroleum ether extract and 15-20 parts of eucommia ethanol extract.
Further preferably, the eucommia ulmoides extract comprises the following components in parts by weight: 39-46 parts of eucommia petroleum ether extract and 16-20 parts of eucommia ethanol extract.
In some preferred embodiments, the ratio of the parts by weight of the eucommia ulmoides petroleum ether extract to the parts by weight of the eucommia ulmoides ethanol extract is 1-5: 1;
preferably, the ratio of the eucommia ulmoides petroleum ether extract to the eucommia ulmoides ethanol extract in parts by weight is 2-4: 1;
further preferably, the ratio of the eucommia ulmoides petroleum ether extract to the eucommia ulmoides ethanol extract in parts by weight is 3: 1.
the eucommia ulmoides extract comprises petroleum ether or ethanol extract of eucommia ulmoides bark, eucommia ulmoides leaf and/or eucommia ulmoides whole plant.
The preparation method of the eucommia ulmoides petroleum ether extract comprises the following steps:
(1) firstly, cleaning eucommia bark, eucommia leaves or whole eucommia plants, drying, ultrasonically soaking for 12-24 hours by using petroleum ether with the volume of 8-10 times, filtering, and collecting filtrate A1 and filter residue a 1;
(2) adding petroleum ether with the volume of 5-6 times that of the filter residue a1 obtained in the step (1) to reflux and extract for 10-15 hours; filtering, collecting filtrate A2, and discarding residue a 2;
(3) and (3) combining the filtrate A1 in the step (1) and the filtrate A2 in the step (2), and carrying out reduced pressure distillation to obtain the eucommia petroleum ether extract.
The volume percentage of the petroleum ether is 60-80%; preferably 70 to 80%, and more preferably 80%.
The temperature of the reflux extraction is 65-80 ℃; preferably 75 deg.c.
The preparation method of the eucommia ethanol extract comprises the following steps:
(1) firstly, cleaning eucommia bark, eucommia leaves or whole eucommia plants, drying, ultrasonically soaking for 12-24 hours by using ethanol with the volume of 8-10 times of that of the eucommia bark, filtering, and collecting filtrate B1 and filter residue B1;
(2) adding 5-6 times of ethanol into the filter residue a1 obtained in the step (1) for reflux extraction for 10-15 hours; filtering, collecting filtrate B2, and discarding residue B2;
(3) and (3) combining the filtrate B1 in the step (1) and the filtrate B2 in the step (2), and distilling under reduced pressure to obtain the eucommia ethanol extract.
The volume percentage of the ethanol is 80-90%; preferably 90%.
The temperature of the reflux extraction is 80-100 ℃; preferably 90 deg.c.
On the other hand, the invention also provides application of the eucommia ulmoides extract in preparing a pharmaceutical composition for treating and preventing osteoporosis.
The pharmaceutical composition comprises the eucommia ulmoides extract and one or more medicinal diluents or carriers.
The eucommia ulmoides extract of the present invention can be used alone or in the form of a pharmaceutical composition.
The pharmaceutical composition can be prepared into raw materials for treating and preventing osteoporosis and conventional pharmaceutical dosage forms, such as tablets, capsules, granules, oral liquid, injection or any other pharmaceutically acceptable dosage forms; can also be used as additive to be added into various beverages and foods to prepare drinks, foods or health products for treating and preventing osteoporosis.
Compared with the prior art, the invention has the beneficial effects that:
(1) in the implementation process, different extraction methods are adopted to extract the effective ingredients of the eucommia ulmoides, so that the effective ingredients of the medicine can be improved to different degrees, and the content of the effective ingredients can be effectively adjusted due to different solubilities of the different effective ingredients in different solvents;
(2) according to the invention, the effect of treating osteoporosis can be effectively improved by controlling the weight part ratio of the eucommia ulmoides petroleum ether extract to the eucommia ulmoides ethanol extract, and the osteoporosis can be treated from the root;
(3) the method for extracting eucommia ulmoides provided by the invention is simple and easy to operate, is beneficial to comprehensive utilization of traditional Chinese medicine components, and reduces medicine waste.
Detailed Description
The present invention is further illustrated by the following specific examples, which are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.
The reagents used in the experimental process are all analytically pure, and the used traditional Chinese medicines are purchased in a drugstore.
Examples 1 to 4 an extract of eucommia ulmoides
Examples | Petroleum ether extract of eucommia ulmoides | Ethanol extract of eucommia ulmoides |
Example 1 | 20 portions of | 10 portions of |
Example 2 | 50 portions of | 30 portions of |
Example 3 | 50 portions of | 10 portions of |
Example 4 | 30 portions of | 30 portions of |
The preparation method comprises the following steps:
the preparation method of the eucommia ulmoides petroleum ether extract comprises the following steps:
(1) firstly, cleaning eucommia bark, eucommia leaves or whole eucommia plants, drying, ultrasonically soaking for 24 hours by using 60 percent petroleum ether with 8 times volume, filtering, and collecting filtrate A1 and filter residue a 1;
(2) adding 60 percent petroleum ether with 5 times volume of the filter residue a1 obtained in the step (1) to perform reflux extraction for 10 hours at the temperature of 80 ℃; filtering, collecting filtrate A2, and discarding residue a 2;
(3) and (3) combining the filtrate A1 in the step (1) and the filtrate A2 in the step (2), and carrying out reduced pressure distillation to obtain the eucommia petroleum ether extract.
The preparation method of the eucommia ethanol extract comprises the following steps:
(1) firstly, cleaning eucommia bark, eucommia leaves or whole eucommia plants, drying, ultrasonically soaking for 24 hours by using 8 times of 80% ethanol, filtering, and collecting filtrate B1 and filter residue B1;
(2) adding 6 times volume of 80% ethanol into the filter residue a1 obtained in the step (1), and performing reflux extraction for 10 hours at 100 ℃; filtering, collecting filtrate B2, and discarding residue B2;
(3) and (3) combining the filtrate B1 in the step (1) and the filtrate B2 in the step (2), and distilling under reduced pressure to obtain the eucommia ethanol extract.
Examples 5 to 8 an extract of eucommia ulmoides
Examples | Petroleum ether extract of eucommia ulmoides | Ethanol extract of eucommia ulmoides |
Example 5 | 30 portions of | 10 portions of |
Example 6 | 39 portions of | 13 portions of |
Example 7 | 45 portions of | 15 portions of |
Example 8 | 48 portions of | 16 portions of |
The preparation method comprises the following steps:
the preparation method of the eucommia ulmoides petroleum ether extract comprises the following steps:
(1) firstly, cleaning eucommia bark, eucommia leaves or whole eucommia plants, drying, ultrasonically soaking for 24 hours by using 10 times of 80% petroleum ether by volume, filtering, and collecting filtrate A1 and filter residue a 1;
(2) adding 6 times of 80% petroleum ether into the filter residue a1 obtained in the step (1) for reflux extraction for 10 hours at 75 ℃; filtering, collecting filtrate A2, and discarding residue a 2;
(3) and (3) combining the filtrate A1 in the step (1) and the filtrate A2 in the step (2), and carrying out reduced pressure distillation to obtain the eucommia petroleum ether extract.
The preparation method of the eucommia ethanol extract comprises the following steps:
(1) firstly, cleaning eucommia bark, eucommia leaves or whole eucommia plants, drying, ultrasonically soaking for 24 hours by using 90% ethanol with 10 times volume, filtering, and collecting filtrate B1 and filter residue B1;
(2) adding 90% ethanol with 6 times volume of the residue a1 obtained in the step (1) to perform reflux extraction for 10 hours at 90 ℃; filtering, collecting filtrate B2, and discarding residue B2;
(3) and (3) combining the filtrate B1 in the step (1) and the filtrate B2 in the step (2), and distilling under reduced pressure to obtain the eucommia ethanol extract.
Comparative example 1
The difference from example 6 is that: the weight ratio of the eucommia petroleum ether extract to the eucommia ethanol extract is 9: 1, 36 parts of eucommia ulmoides petroleum ether extract and 4 parts of eucommia ulmoides ethanol extract, and the other steps and operations are the same as those in example 6.
Comparative example 2
The difference from example 6 is that: the ratio of the eucommia petroleum ether extract to the eucommia ethanol extract in parts by weight is 1: 9, 4 parts of eucommia ulmoides petroleum ether extract and 36 parts of eucommia ulmoides ethanol extract, and the other steps and operations are the same as those in example 6.
Comparative example 3
The difference from example 8 is that: the same procedure as in example 8 was repeated except that 40 parts of petroleum ether extract of eucommia ulmoides was used.
Comparative example 4
The difference from example 8 is that: the same procedure as in example 8 was repeated except that 40 parts of the ethanol extract of eucommia ulmoides was used.
Comparative example 5
The difference from example 8 is that: the petroleum ether content was 50% by volume, and the other steps and operations were the same as in example 8.
Comparative example 6
The difference from example 8 is that: the petroleum ether content was 90% by volume, and the other steps and operations were the same as in example 8.
Comparative example 7
The difference from example 8 is that: the volume percent of ethanol was 70%, and the other steps and operations were the same as in example 8.
Animal experiments
First, the influence of eucommia ulmoides extract on retinoic acid to rat osteoporosis model
Medicine preparation: tretinoin, available from california, guangzhou, biotechnology ltd.
Animals: 170 SD rats, half male and female, were provided by Guangzhou university of traditional Chinese medicine.
Preparing the medicine:
the extracts obtained in the examples 1 to 8 and the comparative examples 1 to 7 are respectively added with 0.5 percent sodium carboxymethyl cellulose solution to be ground into suspensions with certain concentration;
the tretinoin original drug is taken and added with 0.5 percent sodium carboxymethyl cellulose solution to be ground into 1.5g/100mL suspension.
Grouping experiments:
the rats were randomly divided into 17 groups of 10 animals each, a negative control group, a model group, examples 1 to 8 groups and comparative examples 1 to 7 groups.
Animal treatment:
except for the negative control group, rats of other groups are inoculated with intragastric lavage 70mg/kg.d in the morning-1Tretinoin, negative control group, gastric perfusion isovolumetric normal saline, and eucommia ulmoides extracts (10 g/kg. d. calculated according to crude drugs) in the groups of examples 1-8 and comparative examples 1-7-1) Once a day, after 14 days of continuous administration, tretinoin was discontinued, the eucommia ulmoides extracts of examples 1-8 and comparative examples 1-4 were continuously gavaged for 14 days, the rats were subjected to orbital bleeding, and serum phosphorus P, calcium Ca, serum alkaline phosphatase ALP, osteocalcin BGP and rat bone density BMD were measured using the kit, with specific data as shown in table 1 below.
TABLE 1 Effect of eucommia ulmoides extracts obtained in examples 1 to 8 and comparative examples 1 to 7 on femoral Density and biochemical index of blood in rat (mean. + -. standard deviation)
Note: p <0.05, P <0.01
According to the detection data in the table 1, compared with the negative control group, the BMD of the rat in the model group is obviously reduced, the ALP is obviously increased, and both the BMD and the ALP have statistical significance, which indicates that the model making of the rat osteoporosis model is successful. The experimental examples 1 to 8 and comparative examples 1 to 7 showed a certain increase in bone density compared to the model group, the experimental examples 1 to 8 and comparative examples 1 to 7 showed a significant increase in bone density, the examples 1 to 8 showed a significant increase in bone density with statistical significance (P <0.05), the experimental examples 1 to 8 and comparative examples 1 to 7 showed no significant statistical significance in serum phosphorus and serum calcium content compared to the model group, the experimental examples 1 to 8 and comparative examples 1 to 7 showed a significant decrease in ALP compared to the model group, the experimental examples 1 to 8 showed a greater decrease in the level with statistical significance (P <0.05), and the experimental examples 1 to 8 and comparative examples 1 to 7 showed a certain increase in BGP compared to the model group, but showed statistical significance.
Second, the influence of eucommia ulmoides extracts on yang deficiency mouse animal models
Medicine preparation: hydrocortisone, available from Jiuzhou kang Dart Biotech, Inc. of Hubei.
Animals: 340 Kunming mice, male mice, provided by Guangzhou university of traditional Chinese medicine.
Preparing the medicine:
respectively adding 2% Tween-80 solution into the extracts obtained in examples 1-8 and comparative examples 1-7, and grinding to obtain suspension with certain concentration;
grouping experiments:
the rats were randomly divided into 17 groups of 20 animals (10 animals were used for organ index examination and 10 animals were used for swimming time measurement), which were negative control group, model group, examples 1 to 8 groups and comparative examples 1 to 7 groups.
Animal treatment:
(1) influence of eucommia ulmoides extract on immune organ index of yang-deficiency mice
After two days of mouse feeding, except for the negative control group, each group was injected with hydrocortisone at a dose of 25mg/kg for 8 days continuously, and half an hour after each injection, each group of mice was administered with the corresponding drug (15 g/kg/d of the crude drug in examples 1-8 and comparative examples 1-7)-1Administration), the negative control group and the model group were given physiological saline of equal volume, and after administration for half an hour on the ninth day (12 hours without water prohibition),mouse adrenal gland, thymus and spleen were weighed and visceral index calculation was performed, and the results are shown in table 2 below.
(2) Influence of eucommia ulmoides extract on swimming time of yang-deficiency mice
After two days of mouse feeding, except for the negative control group, each group was injected with hydrocortisone at a dose of 25mg/kg for 8 days continuously, and half an hour after each injection, each group of mice was administered with the corresponding drug (15 g/kg/d of the crude drug in examples 1-8 and comparative examples 1-7)-1Administration), the negative control group and the model group were given physiological saline of equal volume, and after administration for half an hour on the ninth day (fasting without water prohibition for 12 hours), the mice were subjected to a swimming test with a load, and the results are shown in table 3 below.
TABLE 2 influence of eucommia ulmoides extracts on adrenal, thymus and spleen indices (mean. + -. standard deviation) of yang-deficient mice
Group of | Adrenal gland index | Index of thymus | Spleen index |
Negative control group | 0.035±0.025 | 0.298±0.042 | 0.386±0.102 |
Model set | 0.013±0.032 | 0.095±0.035 | 0.208±0.105 |
EXAMPLE 1 group | 0.025±0.030** | 0.125±0.026*8 | 0.245±0.095** |
EXAMPLE 2 group | 0.024±0.023** | 0.128±0.305** | 0.236±0.036** |
EXAMPLE 3 group | 0.026±0.045** | 0.128±0.021** | 0.258±0.025** |
EXAMPLE 4 group | 0.025±0.033** | 0.126±0.020** | 0.246±0.056** |
EXAMPLE 5 group | 0.028±0.023** | 0.152±0.014** | 0.279±0.102** |
EXAMPLE 6 group | 0.029±0.052** | 0.156±0.018** | 0.276±0.056** |
EXAMPLE 7 group | 0.029±0.022** | 0.155±0.023** | 0.275±0.045** |
EXAMPLE 8 group | 0.030±0.033** | 0.158±0.024** | 0.280±0.025** |
Comparative example 1 group | 0.022±0.041* | 0.102±0.018* | 0.228±0.046* |
Comparative example 2 group | 0.021±0.055* | 0.105±0.016* | 0.230±0.048* |
Comparative example 3 group | 0.018±0.034 | 0.098±0.021 | 0.220±0.056* |
Comparative example 4 group | 0.019±0.016 | 0.095±0.023 | 0.218±0.064* |
Comparative example 5 group | 0.018±0.022 | 0.115±0.020 | 0.235±0.040 |
Comparative example 6 group | 0.019±0.016 | 0.112±0.022 | 0.238±0.045 |
Comparative example 7 group | 0.022±0.020 | 0.105±0.016 | 0.232±0.048 |
Note: p <0.05, P <0.01
According to the detection data in the table 2, the adrenal gland index, the thymus index and the spleen index of the model group are obviously changed compared with those of the negative control group, which indicates that the model modeling is successful; compared with the model group, the adrenal indexes of the experimental examples 1-8 and the comparative examples 1-7 are increased to a certain extent, the adrenal indexes of the experimental examples 1-8 are obviously increased and have statistical significance (P is less than 0.01), and the adrenal indexes of the comparative examples 1-7 have an increasing trend but have no statistical significance; the thymus index of the experimental examples 1-8 is obviously increased compared with that of the model group and has statistical significance (P is less than 0.01), and the thymus index of the comparative examples 1-7 has a certain increasing trend compared with that of the model group but has no statistical significance; the spleen index and model group were increased for the experimental examples 1-8 and comparative examples 1-7, statistically significant (P <0.05 or P < 0.01).
TABLE 3 influence of eucommia ulmoides extracts on swimming time of yang-deficient mice (mean. + -. standard deviation)
Group of | Swimming time (min) |
Negative control group | 11.25±1.02 |
Model set | 6.45±0.95 |
EXAMPLE 1 group | 9.56±1.03* |
EXAMPLE 2 group | 9.85±0.99* |
EXAMPLE 3 group | 9.46±0.85* |
EXAMPLE 4 group | 8.98±0.96* |
EXAMPLE 5 group | 10.84±1.01** |
EXAMPLE 6 group | 10.69±0.95** |
EXAMPLE 7 group | 10.85±1.05** |
EXAMPLE 8 group | 10.72±0.96** |
Comparative example 1 group | 9.25±1.02* |
Comparative example 2 group | 9.35±0.94* |
Comparative example 3 group | 8.68±1.06* |
Comparative example 4 group | 8.72±0.98* |
Comparative example 5 group | 9.45±0.92* |
Comparative example 6 group | 9.52±0.88* |
Comparative example 7 group | 9.43±0.89* |
Note: p <0.05, P <0.01
As can be seen from the test data in Table 3, the swimming time of the groups of examples 5-8 is significantly higher than that of the model group, and has statistical significance (P <0.01), the swimming time of the groups of examples 1-4 and comparative examples 1-7 is increased (P <0.05) compared with that of the model group, but the useful time is not as long as that of the groups of examples 5-8, which indicates that the pharmaceutical composition provided by the invention can improve the weight bearing swimming time of mice to different degrees.
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 (10)
1. An eucommia ulmoides extract characterized by: the eucommia ulmoides extract comprises the following components in parts by weight: 20-50 parts of eucommia petroleum ether extract and 10-30 parts of eucommia ethanol extract.
2. The eucommia ulmoides extract according to claim 1, wherein: the eucommia ulmoides extract comprises the following components in parts by weight: 30-48 parts of eucommia petroleum ether extract and 15-20 parts of eucommia ethanol extract.
3. The eucommia ulmoides extract according to claim 1, wherein: the weight ratio of the eucommia petroleum ether extract to the eucommia ethanol extract is 1-5: 1.
4. the eucommia ulmoides extract according to claim 1, wherein: the weight ratio of the eucommia petroleum ether extract to the eucommia ethanol extract is 3: 1.
5. the eucommia ulmoides extract according to any one of claims 1 to 4, wherein: the preparation method of the eucommia ulmoides petroleum ether extract comprises the following steps:
(1) firstly, cleaning eucommia bark, eucommia leaves or whole eucommia plants, drying, ultrasonically soaking for 12-24 hours by using petroleum ether with the volume of 8-10 times, filtering, and collecting filtrate A1 and filter residue a 1;
(2) adding petroleum ether with the volume of 5-6 times that of the filter residue a1 obtained in the step (1) to reflux and extract for 10-15 hours; filtering, collecting filtrate A2, and discarding residue a 2;
(3) and (3) combining the filtrate A1 in the step (1) and the filtrate A2 in the step (2), and carrying out reduced pressure distillation to obtain the eucommia petroleum ether extract.
6. The eucommia ulmoides extract according to claim 5, wherein: the volume percentage of the petroleum ether is 60-80%; the temperature of reflux extraction is 65-80 ℃.
7. The eucommia ulmoides extract according to any one of claims 1 to 4, wherein: the preparation method of the eucommia ethanol extract comprises the following steps:
(1) firstly, cleaning eucommia bark, eucommia leaves or whole eucommia plants, drying, ultrasonically soaking for 12-24 hours by using ethanol with the volume of 8-10 times of that of the eucommia bark, filtering, and collecting filtrate B1 and filter residue B1;
(2) adding 5-6 times of ethanol into the filter residue a1 obtained in the step (1) for reflux extraction for 10-15 hours; filtering, collecting filtrate B2, and discarding residue B2;
(3) and (3) combining the filtrate B1 in the step (1) and the filtrate B2 in the step (2), and distilling under reduced pressure to obtain the eucommia ethanol extract.
8. The eucommia ulmoides extract according to claim 7, wherein: the volume percentage of the ethanol is 80-90%; the temperature of reflux extraction is 80-100 ℃.
9. Use of the eucommia ulmoides extract as set forth in any one of claims 1 to 8 for the preparation of a pharmaceutical composition for the treatment and prevention of osteoporosis.
10. The use of claim 9, wherein: the pharmaceutical composition comprising the eucommia ulmoides extract of any one of claims 1 to 8 and one or more pharmaceutically acceptable diluents or carriers.
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