CN107582586A - A kind of application of maidenhair extractive of general flavone in climacteric metancholia is treated - Google Patents
A kind of application of maidenhair extractive of general flavone in climacteric metancholia is treated Download PDFInfo
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- CN107582586A CN107582586A CN201610549084.3A CN201610549084A CN107582586A CN 107582586 A CN107582586 A CN 107582586A CN 201610549084 A CN201610549084 A CN 201610549084A CN 107582586 A CN107582586 A CN 107582586A
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Abstract
The present invention relates to a kind of maidenhair general flavone to treat the application of women's climacteric syndrome medicine, method is after maidenhair herb is crushed, extracted with alcohol reflux, petroleum ether degreasing, then through purification with macroreticular resin, maidenhair general flavone is obtained, is higher than 30% according to the weight content of the general flavone of colorimetric method for determining.Pharmacological testing proves that there is the maidenhair extractive of general flavone stronger treatment climacteric metancholia to act on, and can combine and be applied to as bulk drug in the preparation of anti-climacteric metancholia medicine individually or with other medicines.The purity of general flavone in maidenhair extractive of general flavone prepared by method provided by the invention is high, and preparation method is simple and practical, is adapted to industrialized production；Maidenhair extractive of general flavone provided by the invention can be used for the medicine for preparing treatment climacteric metancholia.Inventor：Cai Decheng, Dou Deqiang.
Description
Technical field
The present invention relates to a kind of autonomic drug extracting method and its application, specifically using maidenhair general flavone as active component
Drug extract and its purposes in climacteric metancholia is treated.
Background technology
Maidenhair (Tagetespatula L.) also known as little marigold, reddish yellow grass, western kind chrysanthemum, smelly chrysanthemum, flannel flower (cloud
South), it is composite family plants of tagetes species, is annual herb, the florescence is July~September.Mexico is originated in, in the middle part of Yunnan Province of China
And there is cultivation the naturalization of the northwestward, central Sichuan and the west and south and WESTERN GUIZHOU, existing China various regions, are a kind of common sights
Admire the beauty of flowers grass.Maidenhair has medicinal and health-care effect, is applied in Lisu, the Yi nationality, distributed over Yunnan, Sichuan and Guizhou as medicine.Its bitter is mild-natured, clearly
Hot dampness removing, cough-relieving.For coughing, dysentery, pertussis, toothache, acute conjunctivitis due to pathogenic wind-fire；It is outer to be used for mumps, breast pain (Jia Min such as China Nationals
Medicine will want [M] China Medical Science Press, 2005.；Zhang Huiyuan Chinese medicine resources will wants [M] Science Presses,
1994.)。
Since the 1980s, people start to study the chemical composition of maidenhair, isolated ten thousand longevity
The Huangs such as chrysanthemum element (patuletin), quercetagetin (quercetagetin), quercetagitrin (quercetagitrin)
Ketone compounds (State Administration of Traditional Chinese Medicine《Chinese book on Chinese herbal medicine》Editorial board, China's book on Chinese herbal medicine [M], Shanghai science tech publishing house,
Version in 1999).Main Ingredients and Appearance is 2- carenes, β-phellandrene, piperitone, piperitenone, terpinolene, lemon in maidenhair volatile oil
(Hu Jianan, the pre-test of maidenhair Chemical Composition of The Essential Oil [J] fragrance flavor and cosmetics, 1992 (01) such as lemon alkene：24-26.；
Romagnoli C, Bruni R, Andreotti E, et al.Chemical characterization and
antifungal activity of essential oil of capitula from wild Indian
Tagetespatula L [J] .Protoplasma, 2005,225 (1-2)：57-65), the multipair maidenhair flowers of people enter at present
Row research, medical value research are less.
Climacteric metancholia (climacteric syndrome, CS), be common women, Yi Fa and multiple gynaecological disease it
One, main clinical manifestation is string dizzy, tinnitus, palpitation and insomnia, warms sweating, irritable, hectic fever or appearance edema of lower extremity, receives
Slow-witted, loose stool or paramenia, irritable emotion etc..As can be seen here, not only show gynecology, also relate to psychiatric department,
The fields such as internal medicine, be it is a kind of easily repeatedly, refractory disease.Conventional Hormone replacement therapy Woman climacteric on modern clinic
Syndrome, it can improve symptom to a certain extent, but also because easily causing a variety of adverse reactions, including initiation uterine cancer, mammary gland
Cancer etc., turn into clinical serious not blunt reaction.Its unique advantage is held in controlling for Chinese medicine pair.Part Chinese medicine can both adjust nanosom
Gland axle endocrine dysfunction, can adjust vegetative nerve functional disturbance again, so as to improve climacteric palpitation and insomnia, warm sweating, tired
The symptom such as hot-tempered irritability, hectic fever or appearance edema of lower extremity, indigestion and loss of appetite, loose stool or paramenia, irritable emotion, improve climacteric women life
Bioplasm amount, has a good application prospect.
Early stage, (Cai Decheng, Dou De are strong, and maidenhairs were total we have found that maidenhair extractive of general flavone has anti-Parkinson effect
Chromocor extract, preparation method and application, application number：201511022920.4 the applying date：2015.12.26).Controlling in the recent period
Treat in climacteric metancholia pharmacodynamics test, we surprisingly have found that maidenhair general flavone also has preferably treatment climacteric comprehensive
Close the pharmacological action of disease.By literature search, there is not maidenhair treatment climacteric metancholia pharmacological action report, it is special then to form this
Profit.The content of general flavone about more than 30% in extractive of general flavone in prepared maidenhair, assay method are (beautiful late referring to document
Newly, the strong Paper mulberry Fruits of Dou De and determination of total flavonoids [J] contemporary Chinese Chinese medicines in feed with paper-mulberry leaf, 2008,11：16-17.).This is specially
For sharp maidenhair through alcohol extracting, purification with macroreticular resin prepares maidenhair general flavone so as to fit being developed into treating climacteric
The medicine of syndrome.
The content of the invention
The technical scheme used to achieve the above object is as follows：
1. establish the method for preparing maidenhair extractive of general flavone
(1) with 80% ethanol water (volume ratio) solution, to maidenhair herb, (including root and aerial part are ground into 10 mesh powder
It is last) Soakage extraction is carried out, each solvent load is 10 times of amounts of quality of medicinal material, and 40 DEG C of temperature are soaked 12 hours, extracted 2 times.Merging carries
Take after liquid through being concentrated to give concentrate (about 1 times of amount of quality of medicinal material)；
(2) the petroleum ether extraction degreasing three times of 1/2 volume of concentrate of above-mentioned concentrate, obtains degreasing concentrate；
(3) further through macroporous absorbent resin HPD100 separation, (resin demand is maidenhair dosage to above-mentioned degreasing concentrate
1/2 times), be first washed with water (5 times of resin volumes), then with 60% ethanol water elute, obtain 60% ethanol water eluent, pass through
It is recovered under reduced pressure ethanol, dry maidenhair extractive of general flavone.
2. maidenhair general flavone is used for climacteric metancholia pharmaceutical research
Above-mentioned maidenhair extractive of general flavone is used for the pharmacological testing research for treating castrated rats climacteric metancholia.
Embodiment
In the present invention, general flavone content is measured using the colorimetric method of following document reports：The strong of Chi Yuxin, Dou De
The assay contemporary Chineses Chinese medicine 2008,10 (11) of Paper mulberry Fruit and general flavone in feed with paper-mulberry leaf：16-17.The brief behaviour of this method
It is as follows to make process：Precision weighs a certain amount of extractive of general flavone, prepares sample solution.Add Al (NO3)3/NaNO2Test solution develops the color,
Using Quercetin as reference substance, colorimetric estimation is carried out at 505nm.According to the absorbance of measure, you can maidenhair is calculated
The content of middle general flavone.
Embodiment 1 prepares maidenhair general flavone
Solvent extraction method and resin adsorption method prepare maidenhair general flavone：
By 1 kilogram of the maidenhair powder that mesh number is 10 mesh, the ethanol water that the volumn concentration of ethanol is 80% soaks
Stain is extracted 2 times, and each solvent load is 10 times of amounts of quality of medicinal material, and 40 DEG C of temperature are soaked 12 hours every time.Merge extract solution, reclaim second
Alcoholic solution, 1 times of about quality of medicinal material amount (about 1 liter) is concentrated into, after 500 milliliters of petroleum ether extractions every time 3 times, obtains degreasing
Solution.HPD100 large pore resin absorption column (about 500 milliliter) of the degreasant solution through having anticipated, successively with water (2500 millis
Rise), 60% ethanol water eluted (2000 milliliters), ethanol is recovered under reduced pressure in 60% ethanol eluate, is dried under reduced pressure
Extractive of general flavone 105g is obtained, it is 34.5g to measure general flavone, and the percentage by weight for accounting for the dried extractive of general flavone is
32.8%, the percentage i.e. yield for accounting for maidenhair raw medicinal material gross weight is 3.45%.
Experimental example 1：Therapeutic action of the maidenhair extractive of general flavone to castrated rats class climacteric metancholia model
1.1 experiment material
1.1.1 animal
Animal：SPF levels, SD rats, female, reproduction age unpregnancy, 200~220g of body weight, have purchased from the long-living biotechnology in Liaoning
Limit company, [SCXK (the Liao Dynasty) 2010-0001].
1.1.2 medicine
The preparation of medicine：Take the maidenhair extractive of general flavone of above-mentioned preparation appropriate, add 0.5% sodium carboxymethylcellulose water
Solution, make to be configured to the solution that concentration is about 10mg/ml, 20mg/ml and 40mg/ml, it is standby.
Injection penicillin powder, HARBIN PHARMACEUTICAL GROUP CO., LTD. General Pharm. Factory, lot number：CA1406017-2；
Female treasure pill, Beijing TongrenTang Co., Ltd Tongrentang Pharmaceutical Factory, lot number 2035165.Specification：Every 100 ball weight
10g.Clinical people's dosage is 50 balls, 2 times a day.Based on body weight 60kg, rat equivalent calculates for 6 times equivalent to people's dosage,
Rat study dosage is 0.823g/kg.
1.1.3 instrument
The multi-functional rat autonomic activities recorders of JD-YLS-1B；KD-204 digital and electronic clinical thermometers；Nikon microscope (day
This NIKON)；400R refrigerated centrifuges (German Heraeus Co., Ltds)；R-911 Full automatics exempt from calculating instrument.
1.2 experimental method
1.2.1 the duplication of castrated rats model
Rat removes bilateral ovaries operation method reference literature, and (Liu Rui connects Zishen Capsules and ovariectomized female rats climacteric is integrated
Experimental study [J] traditional Chinese medicine Leaders of disease model, 2009,06：97-98.).
Healthy reproduction age unpregnancy female rats 60, cleaning grade, body weight 200-220g, it is sham-operation group to leave and take 10 at random
(sham-operation group), remaining rat, which is performed the operation, removes bilateral ovaries：3% amobarbital intraperitoneal injection of anesthesia, abdomen position is fixed, in most end
Under rib, ventrimeson away from cropping at backbone 2cm, sterilization, cut off skin and dorsal muscles, gently press from both sides out ovary, fractionation of fatty layer, ligation
Fallopian tubal, ovary is cut off, with method surgery excision opposite side ovary.When sewing up a wound, injection penicillin powder is spilt in wound
Prevent and treat wound infection.
Postoperative 5th~8 day, vaginal smear examination is carried out to animal daily.Cotton swab is moistened with physiological saline, by cotton swab gently
2~3mm of rat vagina is inserted, makes cotton swab and vagina wall friction 2~3 times, takes out cotton swab, by the withered liquid of vagina on cotton swab equably
It is applied to and carries on the piece of slope, paves, microscopy.Property week residing for judging rat according to the cellular change feature of rat sexual cycle vaginal smear
Stage phase：Preoestrus, the visible a large amount of ellipses of smear have core epithelial cell, a small amount of superficial cell；Estrus, smear
It can be seen that the full irregular superficial cell of visual field profile " fallen leaves shape " accumulation or interconnection are in blocks, there is core epithelial cell on a small quantity
It can be seen that；Metaoestrus, irregular superficial cell in smear, there are core epithelial cell and leucocyte visible；Anoestrum, apply
The visible a large amount of leucocytes of piece and there are core epithelial cell and withered liquid on a small quantity.Animal anestrus, show castration success.
1.2.2 packet and administration
Sham-operation group 10,50 castrated rats are randomly divided into 5 groups：Castration model group, maidenhair general flavone group be high, in,
Low dose group, female treasure pill group.After rat castration 6 weeks, each administration group is according to corresponding dosage gastric infusion, sham-operation group and model
Group gavages the sodium carboxymethyl cellulose solution of equivalent 0.5%, 1 time/d, continuous 4 weeks.
The maidenhair flavone in treating climacteric metancholia each group dosage of table 1
1.2.3 Indexs measure
After the gastric infusion time limit terminates, continuous 4 days, spacious case activity, autonomic activities, the measure of body temperature, measure are carried out respectively
Period is by daily single.
1.2.3.1 case activity measure is opened
The spacious case activity of each group rat test：Spacious case is the interior empty cube that high 40cm, length and width are 80cm.Tank wall surrounding
For black, bottom surface is evenly dividing 25 pieces of grids for equal areas with black line.This experimental observation is carried out in quiet room.
(entered by animal foot in grid and be defined) using animal travels number of squares as horizontal anomalous movement score, using upright number as vertical
Straight activity score (with two forelimbs it is liftoff be a scoring criteria).Indoor sound insulation, keep clean in spacious case before test every time, every time
1, meter records level and Vertical movements number of the rat in spacious case in 5min.
1.2.3.2 autonomic activities determines
The multi-functional rat autonomic activities recorders of JD-YLS-1B are assembled, each group rat are respectively placed in active box, every time
1, read the number of activities of rat in 5min.
1.2.3.3 body temperature (rectal temperature) determines
Rat-rectum temperature is measured with electric body-temperature meter, electronic thermometer is inserted into rat anus about at 2cm, until electronics
Clinical thermometer numerical stability, prompting sound is sent, read data.
1.2.3.4 serum relevant hormone determining
The yellow Jackets 35mg/kg of rats by intraperitoneal injection 3% is anaesthetized, and takes blank vacuum tube pipe, abdominal aortic blood, room temperature
Lower standing, 3000rmp are centrifuged 15 minutes, take serum, and -80 DEG C of refrigerators preserve, and another day determine female two using radio immunoassay
Alcohol (E2), lutropin (LH), follicular stimulating hormone (FSH), gonadotropin-releasing hormone (GRH) (GnRH) are horizontal.
1.2.3.5 Uterine coefficient determines
Put to death after rat extracting blood, uterus is taken out in dissection, and after separating surrounding adhering tissue, electronic analytical balance weighs uterus
Weight, and record data.Calculate organ coefficient：Organ coefficient=uterus weight (mg)/body weight (g) * 100.
1.3 statistical procedures
Through SPSS20.0 statistical softwares processing data, as a result useRepresent, compare between group using single factor test variance point
Analysis, first carries out normality and homogeneity test of variance, and detection meets normality and homogeneity of variance simultaneously, examined with LSD.
1.4 experimental result
1.4.1 maidenhair general flavone opens case activity and the influence of autonomic activities to castrated rats
As a result show, horizontal anomalous movement number and Vertical movements number and autonomic activities of each group rat in spacious case are without bright
Significant difference is different, is shown in Table 2.
The maidenhair general flavone of table 2 opens the influence of case activity to castrated rats
1.4.2 influence of the maidenhair general flavone to castrated rats rectal temperature
As a result show, compared with sham-operation group, rat castration is after 10 weeks, and rectal temperature is remarkably decreased (p ＜ 0.05), with mould
Type group compares, and the high, medium and low dosage group of female treasure pill, general flavone significantly raises rectal temperature (p ＜ 0.01), is shown in Table 3.
Influence of the maidenhair general flavone of table 3 to castrated rats rectal temperature
Note：Compared with sham-operation group, ## represents p ＜ 0.01；With model group ratio, * * represent p ＜ 0.01.
1.4.3 maidenhair general flavone is to 10 weeks rat blood serum E of castration2, the horizontal influence of LH, FSH, GnRH
As a result show, compared with sham-operation group, the change of castration rat model hormone serum level is as follows：E2, GnRH it is notable
Decline (p ＜ 0.01), LH, FSH substantially rise (p ＜ 0.01).Compared with castration model group, female treasure pill group and maidenhair general flavone
Each dosage group E2, GnRH is horizontal obvious rises (p ＜ 0.01 or p ＜ 0.05)；Female treasure pill group and maidenhair general flavone high dose group
LH, FSH level are significantly raised (p ＜ 0.05), though low dose group level has decline, statistics indifference in maidenhair general flavone
It is different, specifically it is shown in Table 4.
The maidenhair general flavone of table 4 is to Ovariectomized Rat Serum E2, LH, FSH, GnRH influence
Note：With sham-operation group ratio, ## represents p ＜ 0.01；With model group ratio, * represents that p ＜ 0.05, * * represent p ＜ 0.01.
1.4.4 influence of the maidenhair general flavone to castrated rats organ coefficient
As a result show, with sham-operation group ratio, castration model group rats Uterine coefficient, be remarkably decreased (p ＜ 0.01)；With castration
Model group rats compare, and the female treasure pill group of positive drug and maidenhair general flavone height, middle dosage substantially rise (p ＜ 0.05 or p ＜
0.01), maidenhair low dose group has risen, but statistics indifference, concrete outcome are shown in Table 5.
Influence of the maidenhair general flavone of table 5 to 10 weeks rat uterus coefficients of castration
Note：With sham-operation group ratio, ## represents p ＜ 0.01；With model group ratio, * represents that p ＜ 0.05, * * represent p ＜ 0.01.
From above Rat-rectum temperature, serum E2, LH, FSH, GnRH be horizontal and the index such as Uterine coefficient, it can be deduced that hole
Sparrow grass general flavone has certain therapeutic action to castrated rats model, i.e. climacteric metancholia.
The specific embodiment of the present invention is above are only, but the design concept of the present invention is not limited thereto, it is all to utilize this structure
Think the change to present invention progress unsubstantiality, the behavior for invading the scope of the present invention all should be belonged to.
Claims (3)
1. a kind of application of maidenhair extractive of general flavone in climacteric metancholia medicine is treated, takes maidenhair herb, use
40%~100% ethanol extracts 1~3 time, and extract solution is concentrated into 1 times of amount of quality of medicinal material, then using 1/2 volume of concentrate
Petroleum ether extraction degreasing three times, the extract solution after degreasing, wave to without petroleum ether taste, then using the macropore of 2 times of extracting liquid volumes
Polymeric adsorbent chromatography, first with the water elution of 5 times of resin volumes, then with 50~95% ethanol elutions to paler colour,
Eluent is collected, is concentrated under reduced pressure, is dried, obtains maidenhair extractive of general flavone, purity is more than 30%.It is characterized in that gained
Maidenhair general flavone, preparing the application in treating climacteric metancholia medicine.
It is 2. as claimed in claim 1, it is characterised in that the extractive of general flavone, which is aided with pharmaceutically conventional auxiliary material, to be prepared into
Various peroral dosage forms, including tablet, oral agents, capsule, granule etc..
3. production method as claimed in claim 1, it is characterised in that the maidenhair general flavone of gained can individually or with it is other
Know and shared with treatment climacteric metancholia function medicament, the treatment for women's climacteric syndrome.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103760259A (en) * | 2014-01-07 | 2014-04-30 | 南开大学 | Indication method for Cd-PCB (Cadmium-Polychlorinated Biphenyl) polluted soil by using maidenhair flowers |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103760259A (en) * | 2014-01-07 | 2014-04-30 | 南开大学 | Indication method for Cd-PCB (Cadmium-Polychlorinated Biphenyl) polluted soil by using maidenhair flowers |
Non-Patent Citations (1)
| Title |
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| 张瑞等: "孔雀草中黄酮类色素最佳提取工艺条件及光、热稳定性的研究", 《食品研究与开发》 * |
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