CN104547306A - Method for preparing parthenocissus dalzielii extract and anti-osteoporosis medicine - Google Patents
Method for preparing parthenocissus dalzielii extract and anti-osteoporosis medicine Download PDFInfo
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Abstract
The invention discloses a method for preparing a parthenocissus dalzielii extract and an anti-osteoporosis medicine. The method comprises the following steps: (1), crushing a parthenocissus dalzielii raw material to obtain raw material powder; (2), carrying out alcohol extraction on the raw material powder; (3), filtering the product obtained from the step (2) to obtain an extract liquid; (4), carrying out first vacuum distillation on the extract liquid to obtain a crude extract liquid; and (5), purifying the crude extract to obtain the parthenocissus dalzielii extract. According to the method, the parthenocissus dalzielii extract can be effectively separated from the natural parthenocissus dalzielii raw material, so as to prepare the anti-osteoporosis medicine which is high in safety and free of hepatotoxicity, so that an effective way is provided for treatment of osteoporosis.
Description
Technical field
The invention belongs to medical domain, specifically, the present invention relates to a kind of method and the osteosporosis resistant medicament of preparing different leaf Radix seu Caulis Parthenocissi tricuspidatae extract.
Background technology
Different leaf Radix seu Caulis Parthenocissi tricuspidatae, is also called different leaf parthenocissus, blade of grass rattan, the Serpentis that climbs the tree, spends vine in vain, is the plant that Vitaceae Radix seu Caulis Parthenocissi tricuspidatae belongs to.Widely distributed, be grown on the area of height above sea level 200 meters to 3800 meters, be often born in cliff cliff, hillside, mountain valley woods and shrubbery rock seam in.There is expelling wind and removing dampness, dredging collateral, hemostasis, Detoxication.Modern pharmacology finds that Vitaceae Radix seu Caulis Parthenocissi tricuspidatae platymiscium contains multiple oligomeric stilbene compound, and natural stilbene compound has antitumor, antiinflammatory, antioxidation, antithrombotic (anticoagulant), blood pressure lowering, blood fat reducing, protect the liver, improve learning and memory function, immunomodulating, the effect such as antibacterial.
Osteoporosis (osteoporosis, OP) be reduce with bone amount, a kind of generalized metabolic osteopathia that the microstructure degeneration of bone is feature, along with the intensification of aged tendency of population, the sickness rate of osteoporosis in middle-aged and elderly people also improves constantly, owing to itself there is no special clinical manifestation, and this disease is again slowly progressive process in development, thus until sickly to send out, always ignore by most people.Osteoporosis disability rate is higher, and extreme influence the quality of life of people, brings heavy financial burden to family and society.Along with population structure aging, osteoporotic problem is more and more outstanding.Osteoporosis is one of most important public health problem of developed country and even developing country, is called silent epidemic diseases.
In whole life cycle, sclerotin, ceaselessly forming (skeletonization) and destroying (broken bone), maintains a kind of dynamic equilibrium.Modern medicine thinks that the basic pathology mechanism of osteoporosis is that bone resorption and osteoplastic coupling occur unbalance in bone metabolism process, causes the alcium and phosphor metabolization in human body uneven, the clinical symptoms that bone density reduces gradually and causes.Clinically desirable method be there is no for the treatment of osteoporosis at present.
Though without osteoporotic disease name in Chinese medicine, study carefully its clinical manifestation, category that pathogenesis should belong to " rheumatism involving the bone ", " atrophic debility of bones ".Theory of Chinese medical science is thought, etiology and pathogenesis mainly deficiency of the kidney, the insufficiency of the spleen and tired resistance arteries and veins of expectorant of osteoporosis.The rule for the treatment of is invigorating the kidney and strengthening the bones, invigorating the spleen and benefiting QI and HuoXueTongLuo oral Liquor medicine are treated osteoporosis, all obtains good therapeutic effect.Therefore anemia treatment medicine is very meaningful safely and effectively to find one.
But the preparation method and its usage at present about different leaf Radix seu Caulis Parthenocissi tricuspidatae extract there is no report.
Summary of the invention
The present invention is intended to solve one of technical problem in correlation technique at least to a certain extent.For this reason, one object of the present invention is that proposition is a kind of and has the method and osteosporosis resistant medicament of preparing different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, and the method effectively can be separated and obtain different leaf Radix seu Caulis Parthenocissi tricuspidatae extract from natural different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material.
In one aspect of the invention, the present invention proposes a kind of method preparing different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, comprising:
(1) different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material is pulverized, to obtain material powder;
(2) described material powder is carried out adding alcohol extraction;
(3) product that step (2) obtains is filtered, to obtain extracting solution;
(4) described extracting solution is carried out the first distilling under reduced pressure, to obtain crude extract; And
(5) described crude extract is carried out purification, to obtain described different leaf Radix seu Caulis Parthenocissi tricuspidatae extract.
Effectively can be separated from natural different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material according to the method for the preparation of the embodiment of the present invention different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and obtain different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, and then it is high and do not have hepatotoxic osteosporosis resistant medicament to prepare safety, thus provide effective way for osteoporotic treatment.
In addition, the method for the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of preparation according to the above embodiment of the present invention can also have following additional technical characteristic:
In some embodiments of the invention, step (2) comprises further: the described material powder alcohol of 75 ~ 95 volume % of 6 ~ 8 times of weight is carried out heating and refluxing extraction 1 ~ 6 time, each 2-3 hour, and is merged by the product obtained respectively.Thus, the separation efficiency of different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can be significantly improved.
In some embodiments of the invention, in step (4), described first distilling under reduced pressure carries out at the temperature of 40 ~ 60 degrees Celsius.Thus, the separation efficiency of different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can be improved further.
In some embodiments of the invention, step (5) comprises further: by described crude extract thin up, to obtain the aqueous solution containing extract; Petroleum ether, chloroform and ethyl acetate is adopted to extract, to obtain petroleum ether extraction liquid, chloroform extraction liquid and acetic acid ethyl acetate extract respectively the described aqueous solution containing extract successively; And respectively remaining aqueous solution after described petroleum ether extraction liquid, described chloroform extraction liquid, described acetic acid ethyl acetate extract and extraction is carried out the second distilling under reduced pressure, to obtain petroleum ether different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, chloroform different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, ethyl acetate different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of aqueous phase respectively.Thus, the separation efficiency of different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can be improved further.
In some embodiments of the invention, described second distilling under reduced pressure carries out at the temperature of 20 ~ 30 degrees Celsius.Thus, the separation efficiency of different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can be improved further.
In some embodiments of the invention, described different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material is the stem of different leaf Radix seu Caulis Parthenocissi tricuspidatae.
In second of the present invention, the present invention proposes a kind of osteosporosis resistant medicament, comprising: different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, described different leaf Radix seu Caulis Parthenocissi tricuspidatae extract is prepared by method described above.Thus, this medicine, by using the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract extracted from natural Chinese medicine different leaf Radix seu Caulis Parthenocissi tricuspidatae, makes its safety high, does not have liver toxicity, and the shortcoming that chemical synthetic drug side effect is large can be avoided, thus provide effective way for osteoporotic treatment.
In addition, osteosporosis resistant medicament according to the above embodiment of the present invention can also have following additional technical characteristic:
In some embodiments of the invention, described osteosporosis resistant medicament comprises adjuvant further.
In some embodiments of the invention, described adjuvant is be selected from least one in solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant, wetting agent, thickening agent, solubilizing agent and substrate.
In some embodiments of the invention, described medicine is powder, granule, tablet, capsule, injection, inhalant or gel.
Additional aspect of the present invention and advantage will part provide in the following description, and part will become obvious from the following description, or be recognized by practice of the present invention.
Accompanying drawing explanation
Fig. 1 is the method flow schematic diagram preparing different leaf Radix seu Caulis Parthenocissi tricuspidatae extract according to an embodiment of the invention;
Fig. 2 is that the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of the embodiment of the present invention is to bone mineral density spectrogram.
Detailed description of the invention
Be described below in detail embodiments of the invention, embodiment described below is exemplary, is intended to for explaining the present invention, and can not be interpreted as limitation of the present invention.
In describing the invention, it is to be appreciated that term " first ", " second " only for describing object, and can not be interpreted as instruction or hint relative importance or the implicit quantity indicating indicated technical characteristic.Thus, be limited with " first ", the feature of " second " can express or impliedly comprise at least one this feature.In describing the invention, the implication of " multiple " is at least two, such as two, three etc., unless otherwise expressly limited specifically.
In one aspect of the invention, the present invention proposes a kind of method preparing different leaf Radix seu Caulis Parthenocissi tricuspidatae extract.Method below with reference to the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of the preparation of Fig. 1 to the embodiment of the present invention is described in detail.According to embodiments of the invention, the method comprises:
S100: different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material is pulverized
According to embodiments of the invention, different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material is pulverized, thus can material powder be obtained.According to embodiments of the invention, different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material is also not particularly limited, and according to a particular embodiment of the invention, different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material can be the stem of different leaf Radix seu Caulis Parthenocissi tricuspidatae.In this step, concrete, after first being dried by the stem of different leaf Radix seu Caulis Parthenocissi tricuspidatae, conducting powder is broken.
S200: material powder is carried out adding alcohol extraction
According to embodiments of the invention, material powder obtained above is added alcohol extraction, thus the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract in material powder can be dissolved in alcohol.
In this step, concrete, the material powder alcohol of 75 ~ 95 volume % of 6 ~ 8 times of weight is carried out heating and refluxing extraction 1 ~ 6 time, each 2-3 hour, and the product obtained respectively is merged.Inventor finds, can significantly improve the separation efficiency of different leaf Radix seu Caulis Parthenocissi tricuspidatae extract under this condition compared with other conditions.It should be noted that, the alcohol that this step uses can be methanol or ethanol.
S300: the above-mentioned product that obtains is filtered
According to embodiments of the invention, the product obtained by S200 filters, thus can obtain extracting solution.Thus, being effectively separated of insoluble matter and extracting solution can be realized.
S400: extracting solution is carried out the first distilling under reduced pressure
According to embodiments of the invention, extracting solution obtained above is carried out the first distilling under reduced pressure, thus can crude extract be obtained.Concrete, extracting solution obtained above is distilled, until obtain extractum shape crude extract.
According to embodiments of the invention, the condition of the first distilling under reduced pressure is also not particularly limited, and according to a particular embodiment of the invention, the first distilling under reduced pressure can be carried out at the temperature of 40 ~ 60 degrees Celsius.Inventor finds, can significantly improve the separation efficiency of different leaf Radix seu Caulis Parthenocissi tricuspidatae extract under this condition.
S500: crude extract is carried out purification
According to embodiments of the invention, crude extract obtained above is carried out purification, thus different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can be obtained.Thus, the purity of different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can be significantly improved.
In this step, concrete, first crude extract is carried out thin up, obtain the aqueous solution containing extract; Then adopt petroleum ether, chloroform and ethyl acetate to extract the aqueous solution containing extract successively, obtain petroleum ether extraction liquid, chloroform extraction liquid and acetic acid ethyl acetate extract respectively; And respectively remaining aqueous solution after petroleum ether extraction liquid, chloroform extraction liquid, acetic acid ethyl acetate extract and extraction is carried out the second distilling under reduced pressure, until extract is concentrated into extractum shape, petroleum ether different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, chloroform different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, ethyl acetate different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of aqueous phase can be obtained respectively.。Thus, the separation efficiency of different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can be improved further.
According to embodiments of the invention, the condition of the second distilling under reduced pressure is also not particularly limited, and according to a particular embodiment of the invention, the second distilling under reduced pressure can be carried out at the temperature of 20 ~ 30 degrees Celsius.Inventor finds, can significantly improve the separation efficiency of different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, thus obtain the higher different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of purity under this condition.
Effectively can be separated from natural different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material according to the method for the preparation of the embodiment of the present invention different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and obtain different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, and then it is high and do not have hepatotoxic osteosporosis resistant medicament to prepare safety, thus provide effective way for osteoporotic treatment.
In second of the present invention, the present invention proposes a kind of osteosporosis resistant medicament.According to embodiments of the invention, this medicine comprises different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, and according to a particular embodiment of the invention, this different leaf Radix seu Caulis Parthenocissi tricuspidatae extract adopts the method for the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of above-mentioned preparation to prepare.Inventor finds, this medicine, by using the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract extracted from natural Chinese medicine different leaf Radix seu Caulis Parthenocissi tricuspidatae, makes its safety high, does not have liver toxicity, and the shortcoming that chemical synthetic drug side effect is large can be avoided, thus provide effective way for osteoporotic treatment.
According to embodiments of the invention, osteosporosis resistant medicament may further include adjuvant.According to a particular embodiment of the invention, adjuvant can for being selected from least one in solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant, wetting agent, thickening agent, solubilizing agent and substrate.
According to embodiments of the invention, medicine can be powder, granule, tablet, capsule, injection, inhalant or gel.Thus, can further for osteoporotic treatment provides effective way.
Below with reference to specific embodiment, present invention is described, it should be noted that, these embodiments are only descriptive, and do not limit the present invention in any way.
Embodiment 1
The stem of different leaf Radix seu Caulis Parthenocissi tricuspidatae is dried and is ground into powder, by the methanol heating and refluxing extraction 3 time of gained material powder with 95 volume % of 6 ~ 8 times of weight, each 2-3 hour, and other product will be divided to merge, then the product obtained is filtered, obtain extracting solution, then extracting solution is carried out distilling under reduced pressure under 40 ~ 60 degrees Celsius, to obtain extractum shape crude extract, then thin up is carried out to crude extract, obtain the aqueous solution containing extract, then petroleum ether is adopted successively, chloroform and ethyl acetate extract the aqueous solution containing extract, obtain petroleum ether extraction liquid respectively, chloroform extraction liquid and acetic acid ethyl acetate extract, then respectively by petroleum ether extraction liquid, chloroform extraction liquid, acetic acid ethyl acetate extract and the rear remaining aqueous solution of extraction carry out distilling under reduced pressure under 20 ~ 30 degrees Celsius, until extract is concentrated into extractum shape and vacuum drying, the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of petroleum ether can be obtained respectively, the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of chloroform, ethyl acetate different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of aqueous phase.
Embodiment 2
The stem of different leaf Radix seu Caulis Parthenocissi tricuspidatae is dried and is ground into powder, by gained material powder with 95% of 6 ~ 8 times of weight alcohol heating reflux extract 4 times, each 2-3 hour, and other product will be divided to merge, then the product obtained is filtered, obtain extracting solution, then extracting solution is carried out distilling under reduced pressure under 40 ~ 60 degrees Celsius, to obtain extractum shape crude extract, then thin up is carried out to crude extract, obtain the aqueous solution containing extract, then petroleum ether is adopted successively, chloroform and ethyl acetate extract the aqueous solution containing extract, obtain petroleum ether extraction liquid respectively, chloroform extraction liquid and acetic acid ethyl acetate extract, then respectively by petroleum ether extraction liquid, chloroform extraction liquid, acetic acid ethyl acetate extract and the rear remaining aqueous solution of extraction carry out distilling under reduced pressure under 20 ~ 30 degrees Celsius, until extract is concentrated into extractum shape and vacuum drying, the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of petroleum ether can be obtained respectively, the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of chloroform, ethyl acetate different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of aqueous phase.
Embodiment 3
The stem of different leaf Radix seu Caulis Parthenocissi tricuspidatae is dried and is ground into powder, by gained material powder with 75% of 6 ~ 8 times of weight alcohol heating reflux extract 6 times, each 2-3 hour, and other product will be divided to merge, then the product obtained is filtered, obtain extracting solution, then extracting solution is carried out distilling under reduced pressure under 40 ~ 60 degrees Celsius, to obtain extractum shape crude extract, then thin up is carried out to crude extract, obtain the aqueous solution containing extract, then petroleum ether is adopted successively, chloroform and ethyl acetate extract the aqueous solution containing extract, obtain petroleum ether extraction liquid respectively, chloroform extraction liquid and acetic acid ethyl acetate extract, then respectively by petroleum ether extraction liquid, chloroform extraction liquid, acetic acid ethyl acetate extract and the rear remaining aqueous solution of extraction carry out distilling under reduced pressure under 20 ~ 30 degrees Celsius, until extract is concentrated into extractum shape and vacuum drying, the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of petroleum ether can be obtained respectively, the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of chloroform, ethyl acetate different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of aqueous phase.
Evaluate:
1, respectively embodiment 1-3 gained crude extract, petroleum ether different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, chloroform different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, ethyl acetate different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of aqueous phase are evaluated drug effect in bone cell proliferation impact and body;
2, evaluation index and method of testing:
A, different leaf Radix seu Caulis Parthenocissi tricuspidatae extract affect bone cell proliferation
1) experimental drug:
Use phosphate buffered solution (PBS) that embodiment 1-3 gained crude extract, petroleum ether different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, chloroform different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, ethyl acetate different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of aqueous phase are mixed with 10mg/ml storing solution respectively, be diluted to 100 μ g/ml before use respectively, 50 μ g/ml, 25 μ g/ml;
2) experimental procedure:
Get the Mus osteoblast (OB) of original cuiture with 2 × 10
4individual/mL is inoculated in 96 orifice plates, and every pore volume 90 μ L, at CO
224h is cultivated in incubator, after cell attachment, add the PBS solution of l0 μ L extract, and make the final concentration of extract remain on 100 μ g/mL respectively, 50 μ g/mL, 25 μ g/mL, after cultivation 48h, every hole adds l0 μ LMTT solution (5mg/mL), continue to hatch 4h in incubator, then incline supernatant, every hole adds DMSO l00 μ L, rear microplate reader of vibrating measures each hole absorbance in 570nm place, set blank group (replacing extract with the PBS of same volume) simultaneously, according to formulae discovery cell survival rate (Cell viability) below=(extract group OD value/blank group OD value) × 100%, experimental result is as shown in table 1.
3) result:
Table 1 embodiment 1-3 extract on bone cell proliferation impact contrast (n=8,
)
Note: compare with matched group, * p < 0.05, * * p < 0.01.
Conclusion: from table 1 data, embodiment 1-3 is compared with matched group, and crude extract, ethyl acetate different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, aqueous phase different leaf Radix seu Caulis Parthenocissi tricuspidatae extract all have obvious proliferation to OB cell under three concentration.
Effect experiment in B, different leaf Radix seu Caulis Parthenocissi tricuspidatae extract body
1) experimental technique:
1.1 dose design and grouping: different leaf Radix seu Caulis Parthenocissi tricuspidatae extract medicine is according to dosage divided into high dose (4g/kg), middle dosage (2g/kg) and low dosage (1g/kg), the matched group that separately designs a model, positive controls and sham-operation negative control group.
1.2 rat modelings
Wistar rat 3v% chloral hydrate normal saline solution anesthesia (dosage is 10mg/kg), conventional method excision bilateral ovaries.Get 10 rats, take out uterus respectively, expose ovary, but do not excise, then put back to abdominal cavity, this group is as sham-operation negative control group.All rats postoperative lumbar injection penicillin (1,600,000 units/bottle penicillin is molten to 5mL), 0.2mL/d, successive administration 7d.
1.3 grouping and administrations
Except sham operated rats, all the other rats were divided into model control group in postoperative 7th day at random by body weight, positive controls, experimental group (experimental group is divided into high dose group, middle dosage group and low dose group by pharmaceutical quantities), often organize 10, the high dose group of experimental group, middle dosage group and low dose group are respectively according to dosage gastric infusion, and positive controls presses 2mg/kg dosed administration, sham-operation negative control group and model control group give the distilled water of Isodose respectively, put to death rat to the 90th day.
2) observation index
2.1 bone densities: get femur stage casing to lower 1/3 place, adopt Dual-energy X-rays absorptionmetry to measure bone density.
2.2 Serum Indexes: after last administration terminates, ventral aorta blood sampling separation of serum, adopts automatic biochemistry analyzer to measure serum levels of ALP content.
2.3 bone histomorphometric parameters: get right side femur, conveniently fix, decalcification, section, HE dyes, adopt multi-functional true color pathological image analyzer under 100 times, under metaphysis growth plate, the visual field is got at the nearly cortex place of 1 ~ 4mm, carry out bone histomorphometric mensuration, calculate bone trabecula number (trabecular number, TN), average bone trabecula thickness (mean trabecular plate thickness, MTPT), Trabecula Bone Volume percentage ratio (trabecular bone volume, and average cortex thickness of bone (mean cortical bone thickness TBV), MCBT).
3) result
3.1 bone density tests: result as shown in Figure 2.
Conclusion: as shown in Figure 2, compare with sham-operation negative control group, model control group bone density significantly reduces (P<0.01), show the osteoporosis rat modeling success that removal ovary causes, compare with model control group, the bone density significance of high dose group, middle dosage group, low dose group raises (P<0.05 or P<0.01), shows that different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can effectively prevent the bone density of osteoporosis rat from reducing.
The mensuration of 3.2 serum levels of ALP content: test result is as shown in table 2.
The different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of table 2 on the impact of Ovariectomized Rats serum levels of ALP content (n=8,
)
Note:
##p<0.01 compares with to sham operated rats;
*p<0.01 compares with model group
Conclusion: from table 2 data, compares with sham-operation negative control group, and in model control group, serum levels of ALP content significantly increases (P<0.01); Compare with model control group, in high dose group, middle dosage group, low dose group and positive controls, serum levels of ALP content all significantly reduces, and shows that different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can reduce serum levels of ALP level.
3.3 bone histomorphometric tests: test result is as shown in table 3.
The different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of table 3 on the impact of Ovariectomized Rats bone histomorphometric (n=8,
)
Note:
##p<0.01 compares with to sham operated rats;
*p<0.05 compares with model group;
*p<0.01 compares with model group;
Conclusion: as shown in Table 3, model control group TN, MTPT, TBV and MCBT and sham-operation negative control group more all have significant difference (P<0.01), show that different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can improve the bone condention index of Osteoporotic Model rat, increase bone amount, compare with model control group, high dose, middle dosage, the bone histomorphometric of low dosage and positive controls all has significant difference, wherein TN increases, MTPT thickens, TBV increases, MCBT increases, show that different leaf Radix seu Caulis Parthenocissi tricuspidatae extract can increase bone formation parameter, reduce bone resorption parameter, effectively can prevent bone loss.
In the description of this description, specific features, structure, material or feature that the description of reference term " embodiment ", " some embodiments ", " example ", " concrete example " or " some examples " etc. means to describe in conjunction with this embodiment or example are contained at least one embodiment of the present invention or example.In this manual, to the schematic representation of above-mentioned term not must for be identical embodiment or example.And the specific features of description, structure, material or feature can combine in one or more embodiment in office or example in an appropriate manner.In addition, when not conflicting, the feature of the different embodiment described in this description or example and different embodiment or example can carry out combining and combining by those skilled in the art.
Although illustrate and describe embodiments of the invention above, be understandable that, above-described embodiment is exemplary, can not be interpreted as limitation of the present invention, and those of ordinary skill in the art can change above-described embodiment within the scope of the invention, revises, replace and modification.
Claims (10)
1. prepare a method for different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, it is characterized in that, comprising:
(1) different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material is pulverized, to obtain material powder;
(2) described material powder is carried out adding alcohol extraction;
(3) product that step (2) obtains is filtered, to obtain extracting solution;
(4) described extracting solution is carried out the first distilling under reduced pressure, to obtain crude extract; And
(5) described crude extract is carried out purification, to obtain described different leaf Radix seu Caulis Parthenocissi tricuspidatae extract.
2. method according to claim 1, is characterized in that, step (2) comprises further:
The described material powder alcohol of 75 ~ 95 volume % of 6 ~ 8 times of weight is carried out heating and refluxing extraction 1 ~ 6 time, each 2-3 hour, and the product obtained respectively is merged.
3. method according to claim 1, is characterized in that, in step (4), described first distilling under reduced pressure carries out at the temperature of 40 ~ 60 degrees Celsius.
4. method according to claim 1, is characterized in that, step (5) comprises further:
By described crude extract thin up, to obtain the aqueous solution containing extract;
Petroleum ether, chloroform and ethyl acetate is adopted to extract, to obtain petroleum ether extraction liquid, chloroform extraction liquid and acetic acid ethyl acetate extract respectively the described aqueous solution containing extract successively; And
Respectively remaining aqueous solution after described petroleum ether extraction liquid, described chloroform extraction liquid, described acetic acid ethyl acetate extract and extraction is carried out the second distilling under reduced pressure, to obtain petroleum ether different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, chloroform different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, ethyl acetate different leaf Radix seu Caulis Parthenocissi tricuspidatae extract and the different leaf Radix seu Caulis Parthenocissi tricuspidatae extract of aqueous phase respectively.
5. method according to claim 4, is characterized in that, described second distilling under reduced pressure carries out at the temperature of 20 ~ 30 degrees Celsius.
6. method according to claim 1, is characterized in that, described different leaf Radix seu Caulis Parthenocissi tricuspidatae raw material is the stem of different leaf Radix seu Caulis Parthenocissi tricuspidatae.
7. an osteosporosis resistant medicament, is characterized in that, comprising:
Different leaf Radix seu Caulis Parthenocissi tricuspidatae extract, described different leaf Radix seu Caulis Parthenocissi tricuspidatae extract is prepared by the method according to any one of claim 1 ~ 6.
8. medicine according to claim 7, is characterized in that, comprises adjuvant further.
9. medicine according to claim 8, is characterized in that, described adjuvant is be selected from least one in solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant, wetting agent, thickening agent, solubilizing agent and substrate.
10. medicine according to claim 8, is characterized in that, described medicine is powder, granule, tablet, capsule, injection, inhalant or gel.
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| JP2005272441A (en) * | 2004-02-25 | 2005-10-06 | National Yang Ming Univ | Dioscoreas sp. extract and its medical use |
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| JP2005272441A (en) * | 2004-02-25 | 2005-10-06 | National Yang Ming Univ | Dioscoreas sp. extract and its medical use |
| CN103976914A (en) * | 2014-05-30 | 2014-08-13 | 广州丹奇日用化工厂有限公司 | China creeper root extracting solution as well as preparation method and application thereof |
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| 丁晓雯等: "《保健食品原理》", 29 February 2008, 西南师范大学出版社 * |
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