CN111643540A - Eucommia ulmoides extract for treating lumbar muscle strain and preparation method thereof - Google Patents
Eucommia ulmoides extract for treating lumbar muscle strain and preparation method thereof Download PDFInfo
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- CN111643540A CN111643540A CN202010598459.1A CN202010598459A CN111643540A CN 111643540 A CN111643540 A CN 111643540A CN 202010598459 A CN202010598459 A CN 202010598459A CN 111643540 A CN111643540 A CN 111643540A
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- eucommia
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- eucommia ulmoides
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Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- 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
-
- 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
- A61K2236/33—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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2601/00—Systems containing only non-condensed rings
- C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
- C07C2601/14—The ring being saturated
Abstract
The invention provides an eucommia ulmoides extract for treating lumbar muscle strain and a preparation method thereof, and belongs to the technical field of traditional Chinese medicine extraction. The eucommia ulmoides extract comprises the following components in parts by weight: 10-20 parts of eucommia ulmoides chlorogenic acid, 15-40 parts of eucommia ulmoides flavone and 20-50 parts of eucommia ulmoides polysaccharide, wherein the extraction method is controlled to respectively extract the active ingredients of the chlorogenic acid, the flavone and the polysaccharide in eucommia ulmoides, so that the active ingredients of the medicine can be released to a greater extent, the waste of the traditional Chinese medicine is reduced, and the weight ratio of the chlorogenic acid to the polysaccharide is controlled to be 1: 1-5, so that the obtained medicinal composition can effectively treat lumbar muscle strain and has a good treatment effect.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine extraction, and particularly relates to an eucommia ulmoides extract for treating lumbar muscle strain and a preparation method thereof.
Background
With the continuous improvement of living standard of people, the mental labor time is greatly increased, and certain civilization diseases such as lumbar muscle strain and the like are natural, thereby seriously affecting the normal life, work and study of people. In daily life, lumbar muscle strain is easily caused by long-term fixed body positions or poor postures. Lumbar muscle strain is mainly caused by the cumulated chronic injury of the soft tissues of the waist, light people cause myofibrosis, heavy people can tear gradually, edema and hemorrhage appear, then fibrosis occurs, and the strain can be buckled into a cord shape or a nearly circular hard knot during local palpation, if the lumbar muscle strain is not treated in time, the strain can be repeatedly attacked, the degeneration of the lumbar vertebra is easily accelerated, ankylosing spondylitis is caused, and the violent symptoms such as the more serious lumbar disc herniation, lumbar spinal stenosis, lumbar spondylolisthesis and the like are caused or aggravated, so that the pain is aggravated, and the treatment difficulty is increased.
Due to the pathological characteristics and treatment method of lumbar muscle strain, the disease is easy to recur and the treatment is troublesome. The Western medicine generally gives anti-inflammatory analgesics, but there are gastrointestinal side effects such as bleeding, etc., and the therapeutic action is not lasting. Simple massage and physical therapy also have the problems of short action time, easy repetition and the like. The traditional Chinese medicine considers that chronic lumbar muscle strain belongs to the category of arthralgia, and is caused by kidney essence loss, malnutrition of tendons and vessels and cold-dampness coagulation collaterals due to overstrain, cold-dampness feeling and senile decline of a long disease. The deficiency of healthy qi and the insecurity of defensive qi are the intrinsic basis of the occurrence of arthralgia syndrome, and the feeling of exogenous pathogenic factors is the external condition of the occurrence of arthralgia syndrome.
At present, various methods for lumbar muscle strain exist, and no specific therapy exists in the day before. Acupuncture is the most traditional and commonly used method, but needs to be performed on acupoints, and some patients are difficult to accept the acupuncture. The electric needle can relieve pain but is only a palliative method. The depth of needle insertion is required to be well controlled by the acupoint injection and the small needle knife, the patient is injured due to the too deep needle insertion of the acupoint injection and the small needle knife, the technical requirements on doctors are high, the effect of monotherapy is not good, and the oral administration and the external application of the traditional Chinese medicine have certain curative effect although the treatment time is long.
Eucommia ulmoides (Eucommiauli Oliv.) is a perennial deciduous tree of eucommia of Eucommiaceae, a unique medicinal plant in China, and is a famous and precious nourishing medicinal material, sweet and slightly pungent in nature and warm in nature and enters liver and kidney meridians. Eucommia bark has been used as a medicine for more than 2000 years, can tonify liver and kidney, treat various chronic lumbar diseases, has the effects of relieving and treating lumbar muscle strain, lumbar hyperosteogeny and soft tissue contusion, and improves waist soreness and lumbago.
For example, chinese patent application 201811018441.9 discloses a method for preparing a capsule medicine for treating lumbar muscle strain, which comprises the following steps: (1) weighing raw materials; (2) mixing pig kidney, fresh radix et caulis Opuntiae Dillenii and fresh fructus Lycii, pulping, adding green salt, mixing, stirring, fermenting, and taking out to obtain fermented pulp; (3) pulverizing Eucommiae cortex, parching, adding fructus Foeniculi, parching, adding into water, boiling with strong fire, adding fermented pulp, corn pollen and pulverized pericarpium Citri Tangerinae, stirring, boiling with slow fire, adding green salt, stirring, and decocting with slow fire to obtain decoction; (4) magnetizing the decoction, ultrasonic treating, filtering, collecting filtrate, spray drying, adding corm Eleocharitis extract and caulis Sargentodoxae extract, stirring, and making into capsule; has the advantages of quick response, high cure rate, no toxic or side effect, safety and convenience for treating lumbar muscle strain. The capsule medicine is added with eucommia bark, but the capsule medicine uses the original medicine as the medicine component, has long treatment time and is easy to relapse.
As another example, chinese patent application 201310250407.5 discloses a pharmaceutical composition for treating muscle strain, which is a preparation prepared from the following raw materials by weight: 26-34 g of parasitic loranthus, 12-18 g of teasel root, 12-18 g of radix angelicae pubescentis, 12-18 g of eucommia ulmoides, 12-18 g of rhizoma cibotii, 40-50 g of radix paeoniae alba, 15-20 g of achyranthes bidentata, 15-18 g of radix clematidis, 7-13 g of cortex acanthopanacis, 7-13 g of radix saposhnikoviae, 15-20 g of radix puerariae, 15-20 g of pawpaw, 24-30 g of caulis spatholobi, 10-15 g of rhizoma corydalis, 3-7 g of pollen typhae, 7-13 g of frankincense, 7-13 g of myrrh, 10-15 g of rhizoma cyperi and 10. The pharmaceutical composition also uses crude drugs as pharmaceutical components, has long treatment time and is easy to relapse.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims at an eucommia ulmoides extract for treating lumbar muscle strain and a preparation method thereof.
In one aspect, the invention provides an eucommia ulmoides extract, which comprises the following components in parts by weight: 10-20 parts of eucommia chlorogenic acid, 15-40 parts of eucommia flavone and 20-50 parts of eucommia polysaccharide.
Preferably, the eucommia ulmoides extract comprises the following components in parts by weight: 12-18 parts of eucommia chlorogenic acid, 20-35 parts of eucommia flavone and 30-45 parts of eucommia polysaccharide.
Still preferably, the eucommia ulmoides extract comprises, by weight: 14-16 parts of eucommia chlorogenic acid, 25-30 parts of eucommia flavone and 35-40 parts of eucommia polysaccharide.
In some preferred embodiments, the weight fraction ratio of eucommia chlorogenic acid to eucommia polysaccharide is 1: 1-5; preferably 1: 1.8-3.75; more preferably 1: 2.2-2.8; more preferably 1: 2.5.
on the other hand, the invention also provides a preparation method of the extract, which comprises the following steps:
method for extracting eucommia ulmoides chlorogenic acid
S1-1, drying folium cortex eucommiae, crushing, adding petroleum ether, performing ultrasonic extraction, repeating the operation for 1-2 times, and filtering and separating filtrate and filter residue;
s1-2, soaking the filter residue obtained in the step S1-1 in ethanol, performing reflux extraction, filtering, and performing reduced pressure distillation on the filtrate to obtain a eucommia crude extract;
s1-3, dissolving the eucommia crude extract obtained in the S1-2 in ethanol, stirring, taking supernatant, concentrating the supernatant under reduced pressure to obtain extract, adding water to the extract to dissolve the extract, passing through macroporous resin, eluting with methanol to obtain eluent, and spray-drying the eluent to obtain the eucommia chlorogenic acid.
The ultrasonic temperature in the step S1-1 is 40-45 ℃, 100-120Hz, and the ultrasonic time is 1-2 hours.
The volume fraction of the ethanol in the step S1-2 is 80-90%, preferably 90%, and the reflux temperature is 70-90 ℃, preferably 80 ℃; the reflux time is 1-2 hours.
The macroporous resin used in the S1-3 is S-8 resin, the pH is 2-3, and the volume fraction of the methanol is 70-80%, preferably 70%.
The extraction method of the eucommia ulmoides flavonoids comprises the following steps:
s2-1, drying folium Eucommiae, pulverizing, extracting with ethanol under reflux for 1-3 times, and mixing extractive solutions;
s2-2, concentrating the extracting solution obtained in the step S2-1 under reduced pressure to obtain a concentrated solution, passing the concentrated solution through macroporous resin, and eluting with ethanol to obtain the eucommia ulmoides flavone extract.
The volume fraction of hexanol described in the above step S2-1 is 70 to 80%, preferably 80%; the reflux temperature is 70-80 deg.C, preferably 80 deg.C, and the reflux time is 2-3 hr.
The macroporous resin used in the step S2-2 is a macroporous resin prepared from the following raw materials in a volume ratio of 1: 1, and X-5 resin, the volume fraction of ethanol used being 70-80%, preferably 75%.
The extraction method of the eucommia polysaccharide comprises the following steps:
s3-1, drying folium Eucommiae, pulverizing, soaking in ethanol, reflux-extracting for 1-3 times, and mixing extractive solutions;
s3-2, concentrating the extracting solution obtained in the step S3-1 under reduced pressure to obtain a concentrated solution, passing the concentrated solution through macroporous resin, and eluting with ethanol to obtain the eucommia polysaccharide extract.
The volume fraction of hexanol described in the above step S3-1 is 60 to 70%, preferably 70%; the reflux temperature is 80-90 deg.C, preferably 90 deg.C, and the reflux time is 2-3 hr.
The macroporous resin used in the step S3-2 is a macroporous resin prepared from the following raw materials in a volume ratio of 1: 1 XDA-1 and X-5 resin, with a volume fraction of ethanol used of 30 to 40%, preferably 40%.
On the other hand, the invention also provides application of the eucommia ulmoides extract in preparing a pharmaceutical composition for treating and preventing lumbar muscle strain.
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 lumbar muscle strain 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 beverages, foods or health products for treating and preventing lumbar muscle strain.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention respectively extracts the effective components of chlorogenic acid, flavone and polysaccharide in eucommia bark by controlling an extraction method, and controls the weight fraction ratio of chlorogenic acid polysaccharide to be 1: 1-5, preparing a pharmaceutical composition which can effectively treat lumbar muscle strain and has good treatment effect;
(2) according to the invention, by controlling and respectively extracting the effective components in the eucommia ulmoides, the effective components of the medicine can be released to a greater extent, and the waste of the traditional Chinese medicine is reduced, so that the problem of low utilization rate of the traditional Chinese medicine components is solved to a certain extent;
(3) the extracted traditional Chinese medicine components can be continuously recycled through subsequent treatment, so that multiple utilization of the traditional Chinese medicine components is realized, and medicine waste is avoided.
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 for the treatment of lumbar muscle strain
| Examples of the invention | Eucommia ulmoides chlorogenic acid | Eucommia ulmoides flavone | Eucommia ulmoides polysaccharide |
| Example 1 | 10 portions of | 15 portions of | 20 portions of |
| Example 2 | 18 portions of | 35 portions of | 45 portions of |
| Example 3 | 16 portions of | 30 portions of | 40 portions of |
| Example 4 | 20 portions of | 40 portions of | 50 portions of |
The preparation method of the extract comprises the following steps:
method for extracting eucommia ulmoides chlorogenic acid
S1-1, drying folium Eucommiae, pulverizing, adding petroleum ether, performing ultrasonic extraction at 40 deg.C under 100Hz, repeating the operation for 2 times, filtering, and separating filtrate and residue;
s1-2, soaking the filter residue obtained in the step S1-1 in 80% ethanol, refluxing at 70 ℃ for 2 hours, filtering, and distilling the filtrate under reduced pressure to obtain a eucommia crude extract;
s1-3, dissolving the eucommia crude extract obtained in the S1-2 in 80% ethanol, stirring, taking supernatant, concentrating the supernatant under reduced pressure to obtain extract, adding water to the obtained extract to dissolve the extract, passing the dissolved extract through S-8 macroporous resin, controlling the pH to be 2, eluting the extract with 70% methanol to obtain eluent, and performing spray drying on the eluent to obtain eucommia chlorogenic acid.
The extraction method of the eucommia ulmoides flavonoids comprises the following steps:
s2-1, drying folium Eucommiae, pulverizing, adding 70% ethanol, refluxing at 70 deg.C for 3 hr, reflux extracting for 3 times, and mixing extractive solutions;
s2-2, concentrating the extracting solution obtained in the step S2-1 under reduced pressure to obtain a concentrated solution, and treating the concentrated solution with a solvent of 1: 1 XDA-1 and X-5 macroporous resin, eluting with 70% ethanol to obtain Eucommiae cortex flavone extract.
The extraction method of the eucommia polysaccharide comprises the following steps:
s3-1, drying folium Eucommiae, pulverizing, soaking in 60% ethanol, reflux-extracting at 80 deg.C for 2 hr for 3 times, and mixing extractive solutions;
s3-2, concentrating the extracting solution obtained in the step S3-1 under reduced pressure to obtain a concentrated solution, and treating the concentrated solution with a solvent of 1: 1 XDA-1 and X-5 macroporous resin, eluting with 30% ethanol to obtain Eucommiae cortex polysaccharide extract.
The preparation method of the pharmaceutical composition comprises the following steps:
mixing the prepared chlorogenic acid, flavone and polysaccharide extracts according to the content of the examples 1-4 to obtain the pharmaceutical compositions A-D.
Examples 5 to 8 an extract of eucommia ulmoides for the treatment of lumbar muscle strain
| Examples of the invention | Eucommia ulmoides chlorogenic acid | Eucommia ulmoides flavone | Eucommia ulmoides polysaccharide |
| Example 5 | 20 portions of | 30 portions of | 20 portions of |
| Example 6 | 10 portions of | 20 portions of | 50 portions of |
| Example 7 | 15 portions of | 35 portions of | 27 portions of |
| Example 8 | 18 portions of | 40 portions of | 45 portions of |
The preparation method of the extract comprises the following steps:
method for extracting eucommia ulmoides chlorogenic acid
S1-1, drying folium Eucommiae, pulverizing, adding petroleum ether, ultrasonic extracting at 45 deg.C under 120Hz for 2 times, filtering, and separating filtrate and residue;
s1-2, adding 90% ethanol into the filter residue obtained in the step S1-1 for soaking, refluxing for 2 hours at 80 ℃, filtering, and distilling the filtrate under reduced pressure to obtain a eucommia crude extract;
s1-3, dissolving the eucommia crude extract obtained in the S1-2 in 90% ethanol, stirring, taking supernatant, concentrating the supernatant under reduced pressure to obtain extract, adding water to the obtained extract to dissolve the extract, passing the dissolved extract through S-8 macroporous resin, controlling the pH to be 2, eluting the extract with 70% methanol to obtain eluent, and performing spray drying on the eluent to obtain eucommia chlorogenic acid.
The extraction method of the eucommia ulmoides flavonoids comprises the following steps:
s2-1, drying folium Eucommiae, pulverizing, adding 80% ethanol, refluxing at 80 deg.C for 3 hr, reflux extracting for 3 times, and mixing extractive solutions;
s2-2, concentrating the extracting solution obtained in the step S2-1 under reduced pressure to obtain a concentrated solution, and treating the concentrated solution with a solvent of 1: 1 XDA-1 and X-5 macroporous resin, eluting with 75% ethanol to obtain Eucommiae cortex flavone extract.
The extraction method of the eucommia polysaccharide comprises the following steps:
s3-1, drying folium Eucommiae, pulverizing, soaking in 70% ethanol, refluxing at 90 deg.C for 2 hr, reflux-extracting for 3 times, and mixing extractive solutions;
s3-2, concentrating the extracting solution obtained in the step S3-1 under reduced pressure to obtain a concentrated solution, and treating the concentrated solution with a solvent of 1: 1 XDA-1 and X-5 macroporous resin, eluting with 40% ethanol to obtain Eucommiae cortex polysaccharide extract.
The preparation method of the pharmaceutical composition comprises the following steps:
mixing the prepared chlorogenic acid, flavone and polysaccharide extracts according to the content of the examples 5-8 to obtain the pharmaceutical composition E-H.
Comparative examples 1 to 5 eucommia ulmoides extract for treating lumbar muscle strain
| Examples of the invention | Eucommia ulmoides chlorogenic acid | Eucommia ulmoides flavone | Eucommia ulmoides polysaccharide |
| Comparative example 1 | - | 40 portions of | 45 portions of |
| Comparative example 2 | 18 portions of | - | 45 portions of |
| Comparative example 3 | 18 portions of | 40 portions of | - |
| Comparative example 4 | 52.5 parts | 40 portions of | 10.5 portions |
| Comparative example 5 | 7 portions of | 40 portions of | 56 portions of |
The extracts described in comparative examples 1 to 5 were prepared in the same manner as described in examples 5 to 8.
Comparative example 6
The difference from example 7 is that: in steps S2-2 and S3-2, the concentrated solution was passed through XDA-1 macroporous resin, and the other operations were the same as in example 7.
Comparative example 7
The difference from example 7 is that: in steps S2-2 and S3-2, concentrate 3: 1 XDA-1 and X-5 macroporous resin, the other operations were the same as in example 7.
Comparative example 8
The difference from example 7 is that: in steps S2-2 and S3-2, concentrate 1: 3 XDA-1 and X-5 macroporous resin, the other operations were the same as in example 7.
The preparation method of the pharmaceutical composition comprises the following steps:
mixing the prepared chlorogenic acid, flavone and polysaccharide extracts according to the content of comparative examples 1-8 to obtain the pharmaceutical composition I-P.
Animal experiments
Toxicity test
1. Acute toxicity test
Experimental animals: the Kunming mouse with the weight of 22-25g and half of the male and female, the drug extract prepared by the embodiment and the comparative example of the invention is prepared into the required test solution by distilled water;
the experimental conditions are as follows: the temperature is 26-28 ℃, and the humidity is 42-46%;
the experimental method comprises the following steps:
animal grouping: each example and comparative example set two dose groups of 10 mice each, half a female and half a male;
dose and mode of administration: two dosage groups of 150mg/kg and 350mg/kg are set for experiments, oral gavage is adopted for administration, the gavage is carried out 3 times every day, 10 days are continuously observed, and the experimental results are shown in table 1:
TABLE 1 toxicity test results in mice
As can be seen from Table 1, no toxic symptoms and death of experimental mice appear in the observation period, the eucommia ulmoides extract for treating lumbar muscle strain has no toxic effect on the mice, and the acute toxicity of the eucommia ulmoides extract belongs to the actual nontoxic class.
2. Long term toxicity test
After the pharmaceutical compositions E-H prepared in examples 5-8 of the present invention were continuously administered to mice for 16 weeks (300mg/kg body weight, 2 times a day) and stopped for 4 weeks, the results showed that: the traditional Chinese medicine composition has no obvious influence on indexes such as hair, behaviors, excrement and urine, body weight, weight of organs, hemogram, liver and kidney functions, blood sugar, blood fat and the like of a mouse, and the macroscopic change of the organs and histological examination results show that after the traditional Chinese medicine composition is taken for 16 weeks and stopped for 4 weeks, all organs of the mouse have no obvious change. The pharmaceutical composition has low toxicity after long-term administration to mice, has no abnormal reaction after drug withdrawal, and is safe to apply.
3. Influence on autophagy-related protein Beclin1 in rat lumbar multifidus muscle injury repair process
Experimental animals: 300 male SD rats with weight of 280-310g are randomly bred in cages with light and shade period of 12 hours, and are bred adaptively for 7 days with free diet, room temperature of 24 ℃ and humidity of 40-50%.
Animal grouping: 216 rats were divided into a blank group, a model group, examples 1 to 8 groups and comparative examples 1 to 8 groups by a random method, 12 rats were collected in each group, and 4 rats were collected at the same time point (8 am) on days 1, 3, 5, and 7.
Animal molding: 10% chloral hydrate (350mg/kg) is used for abdominal injection anesthesia, skin preparation is carried out on the back, 4 points on the two sides of the 4 th and 5 th lumbar vertebra levels of the spine are selected, 0.5% bupivacaine is extracted and intramuscular injected into the selected 4 points on the two sides of the 4 th and 5 th lumbar vertebra levels, 100 mu L of injection is carried out on each point, and a needle head is ensured to reach the multifidus muscle, so that the complete molding is carried out. The blank groups were not processed.
The treatment method comprises the following steps:
blank group: the rats are fed together under the same conditions as the rats in other groups, without molding treatment and drug feeding, and are synchronously taken at the same time points (8 am) on the 1 st, 3 rd, 5 th and 7 th days after molding.
Model group: no treatment was performed, and the material was taken simultaneously with other rats at the same time points (8 am) on days 1, 3, 5 and 7 after the model was created.
Examples 1-8 groups: the pharmaceutical compositions A-H were administered 3 times a day (300mg/kg each time) from day 2 after molding, and were obtained at the same time (8 am) on days 1, 3, 5, and 7 after treatment.
Comparative examples 1-5 groups: the pharmaceutical composition I-M is administered 3 times a day (300mg/kg each time) from day 2 after molding, and is obtained at the same time (8 am) on days 1, 3, 5, and 7 after treatment.
Statistical analysis was performed using SPSS 20.0 software, and statistical data are expressed as mean ± standard deviation.
TABLE 2 comparison of MOD values expressed by the multifidus muscle Beclin1 at different periods in the rats (optical density values, mean. + -. standard deviation)
Note: in comparison to the blank set, the data is,ap is less than or equal to 0.01; in comparison to the set of models,bP≤0.01
according to the detection data in the table 2, the blank group is not treated, the expression level of Beclin1 is lower, the expression level of Beclin1 in the model group, the groups of examples 1-8 and the groups of comparative examples 1-8 is increased (P is less than or equal to 0.01) compared with the blank group after 1 day of treatment, the expression level of Beclin1 in the groups of examples 1-8 is slightly increased compared with the model group, and the expression level of Beclin1 in the groups of comparative examples 1-8 is equivalent to that of the model group, so that the statistical significance is not achieved; after 3 days of treatment, the expression level of Beclin1 in the model group and the groups of examples 1-8 was increased (P.ltoreq.0.01), the expression level of Beclin1 in the groups of comparative examples 1-8 was increased less than that in the groups of examples 1-8, the expression level of Beclin1 in the groups of examples 1-8 was increased (P.ltoreq.0.01), and the expression level of Beclin1 in the groups of comparative examples 1-8 was increased less than that in the groups of examples 1-8; after 5 days of treatment, the expression level of Beclin1 in the model group and the groups of examples 1-8 was increased (P.ltoreq.0.01), the expression level of Beclin1 in the groups of comparative examples 1-8 was increased less than that in the groups of examples 1-8, the expression level of Beclin1 in the groups of examples 1-8 was increased (P.ltoreq.0.01), and the expression level of Beclin1 in the groups of comparative examples 1-8 was increased less than that in the groups of examples 1-8; after 7 days of treatment, the expression level of Beclin1 in the model group and the groups of examples 1-8 is increased (P is less than or equal to 0.01) compared with that in the blank group, the expression level of Beclin1 in the groups of comparative examples 1-5 is increased less than that in the groups of examples 1-8, the expression level of Beclin1 in the groups of examples 1-8 is increased (P is less than or equal to 0.01) compared with that in the model group, and the expression level of Beclin1 in the groups of comparative examples 1-8 is increased less than that in the groups of examples 1-8, which shows that the pharmaceutical compositions prepared in the groups of examples 1-8 can better repair rat lumbar multifidus muscle injury.
4. Clinical trial
390 patients were randomly selected, 200 women and 220 men. 10 days before the experiment, the population was continuously observed, and symptoms and changes were recorded and randomly divided into 13 groups, examples 5-8 groups, comparative examples 1-8 groups and a control group, examples 5-8 groups were administered with the pharmaceutical compositions E-H, comparative examples 1-8 groups were administered with the pharmaceutical compositions I-P, and the control group was administered with the Yaotongning capsules. The taking method comprises the following steps: the preparation is taken orally 3 times a day, 15g each time, and 10 days as a treatment course, wherein the preparation is taken once in the morning, in the middle of the day and at night.
The judgment standard of the curative effect condition is as follows: and (3) curing: after the product is taken for one treatment course, the waist of a patient can move freely, the stabbing pain of the waist can be completely eliminated, and the uncomfortable symptoms can be eliminated; the method has the following advantages: after the product is taken for a course of treatment, discomfort symptoms are relieved or partial discomfort symptoms disappear, and pain is relieved; and (4) invalidation: after the product is taken for a course of treatment, discomfort symptoms are unchanged. The results are shown in Table 2.
TABLE 3 verification of Effect after taking pharmaceutical compositions
As can be seen from table 3 above, the therapeutic effect of the pharmaceutical composition provided by the present invention is significantly higher than that of the control group, the total effective rate of the pharmaceutical composition H obtained in example 8 of the present invention is as high as 96.7%, the reduction of a certain component in comparative examples 1 to 3 significantly reduces the pharmaceutical effect and the cure rate, the change of the quality of the pharmaceutical component in comparative examples 4 to 5 also has a certain effect on the pharmaceutical effect, but the therapeutic effect is inferior to that of the pharmaceutical composition prepared in examples 5 to 8, and the change of the kind of column in the preparation process in comparative examples 6 to 8 also has an effect on the component of the pharmaceutical extract and has a certain effect on the pharmaceutical effect.
It should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and the present invention is not limited thereto, and although the present invention is described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications and equivalents can be made in the technical solutions described in the foregoing embodiments, or some technical features of the present invention may be substituted. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in 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: 10-20 parts of eucommia chlorogenic acid, 15-40 parts of eucommia flavone and 20-50 parts of eucommia polysaccharide.
2. The eucommia ulmoides extract according to claim 1, wherein: the eucommia ulmoides extract comprises the following components in parts by weight: 12-18 parts of eucommia chlorogenic acid, 20-35 parts of eucommia flavone and 30-45 parts of eucommia polysaccharide.
3. The eucommia ulmoides extract according to claim 1, wherein: the eucommia ulmoides extract comprises the following components in parts by weight: 14-16 parts of eucommia chlorogenic acid, 25-30 parts of eucommia flavone and 35-40 parts of eucommia polysaccharide.
4. The eucommia ulmoides extract according to claim 1, wherein: the weight ratio of the eucommia chlorogenic acid to the eucommia polysaccharide is 1: 1-5.
5. The eucommia ulmoides extract according to claim 4, wherein: the weight ratio of the eucommia chlorogenic acid to the eucommia polysaccharide is 1: 2.5.
6. the method for preparing an eucommia ulmoides extract according to any one of claims 1 to 5, wherein: the method comprises the following steps:
(1) method for extracting eucommia ulmoides chlorogenic acid
S1-1, drying folium cortex eucommiae, crushing, adding petroleum ether, performing ultrasonic extraction, repeating the operation for 1-2 times, and filtering and separating filtrate and filter residue;
s1-2, soaking the filter residue obtained in the step S1-1 in ethanol, performing reflux extraction, filtering, and performing reduced pressure distillation on the filtrate to obtain a eucommia crude extract;
s1-3, dissolving the eucommia crude extract obtained in the S1-2 in ethanol, stirring, taking supernatant, concentrating the supernatant under reduced pressure to obtain extract, adding water to the extract to dissolve the extract, passing through macroporous resin, eluting with methanol to obtain eluent, and spray-drying the eluent to obtain the eucommia chlorogenic acid.
(2) The extraction method of the eucommia ulmoides flavonoids comprises the following steps:
s2-1, drying folium Eucommiae, pulverizing, extracting with ethanol under reflux for 1-3 times, and mixing extractive solutions;
s2-2, concentrating the extracting solution obtained in the step S2-1 under reduced pressure to obtain a concentrated solution, passing the concentrated solution through macroporous resin, and eluting with ethanol to obtain the eucommia ulmoides flavone extract.
(3) The extraction method of the eucommia polysaccharide comprises the following steps:
s3-1, drying folium Eucommiae, pulverizing, soaking in ethanol, reflux-extracting for 1-3 times, and mixing extractive solutions;
s3-2, concentrating the extracting solution obtained in the step S3-1 under reduced pressure to obtain a concentrated solution, passing the concentrated solution through macroporous resin, and eluting with ethanol to obtain the eucommia polysaccharide extract.
7. The method of claim 6, wherein: the macroporous resin used in the step S1-3 is S-8 resin, the pH is 2-3, and the volume fraction of the methanol is 70-80%.
8. The method of claim 6, wherein: the macroporous resin used in the step S2-2 is prepared from the following components in a volume ratio of 1: 1 XDA-1 and X-5 resin, the volume fraction of ethanol used is 70-80%;
the macroporous resin used in the step S3-2 is prepared from the following components in a volume ratio of 1: 1 XDA-1 and X-5 resin, with a volume fraction of ethanol used of 30-40%.
9. Use of the eucommia ulmoides extract according to any one of claims 1 to 5 and the extract prepared according to any one of claims 6 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 comprises the eucommia ulmoides extract of any one of claims 1 to 5, the extract prepared by the method of claims 6 to 8 and one or more pharmaceutically acceptable diluents or carriers.
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