CN102210800A - New application of citron daylily and citron daylily general flavone - Google Patents
New application of citron daylily and citron daylily general flavone Download PDFInfo
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Abstract
The invention relates to new application of citron daylily and citron daylily general flavone, and discloses application of citron daylily and citron daylily general flavone to the preparation of medicaments for resisting depression, an particularly discloses application of citron daylily and citron daylily general flavone to the preparation of medicaments for rising serotonin, norepinephrine, dopamine and 5-hydroxyindolacetic acid in cerebral cortex, and discloses application of citron daylily and citron daylily general flavone to the preparation of medicaments for antagonistic reserpine. The application is verified by pharmacological experiments.
Description
The present invention is for dividing an application, and the original bill application number is 200810135327.4, and the original bill applying date is on July 31st, 2008, and the original bill name is called the new purposes of Radix hemerocalis plicatae and Radix hemerocalis plicatae total flavonoids.
Technical field
The present invention relates to the new purposes of a kind of Chinese medicine and effective site thereof, particularly Radix hemerocalis plicatae and the new purposes of Radix hemerocalis plicatae total flavonoids and preparation method thereof.
Background technology
Radix hemerocalis plicatae has another name called " Flos Hemerocallis ", " Bulbus Lilii on the one ", is commonly called as and forgets the melancholy grass.Belong to the Liliaceae day lily, have abundant nutrition.Its main chemical compositions has protein, fat, carbohydrate, also has multivitamin, alkaloid, glycosides, flavonoid, carotene, volatile oil, tannin and inorganic mineral calcium, phosphorus.Contain alkaloid 0.7%, glycosides 1.8%, flavonoid 0.75%, beta-carotene 0.36% according to " Chinese medicine voluminous dictionary " record Radix hemerocalis plicatae.The effect that Ming Dynasty's Li Shizhen (1518-1593 A.D.) claims it that tranquilization and refreshment is arranged in Compendium of Material Medica, increases the intelligence the chest stuffiness relieving, improves looks and nourish blood, separate heat sterilization, relieving restlessness lactogenesis.
The modern study Radix hemerocalis plicatae has effects such as the strong intelligence of supplementing the brain, removing stasis to stop bleeding, removing obstruction in the collateral to relieve pain, anti-inflammation.Wherein Radix hemerocalis plicatae total flavonoids is one of its important effective ingredient, and it has enhancing immunity, blood circulation promoting and blood stasis dispelling, protects the liver antiinflammatory, anti-bacteria and anti-virus, inhibition tumor, remove multiple functions such as oxygen-derived free radicals and bacteriostasis.At present the extraction of Radix hemerocalis plicatae total flavonoids is studied seldom, and its applied research is not appeared in the newspapers.
Summary of the invention
First purpose of the present invention has been to disclose the new purposes of Radix hemerocalis plicatae.
Second purpose of the present invention is the new purposes of open Radix hemerocalis plicatae total flavonoids.
The 3rd purpose of the present invention is the preparation method of open Radix hemerocalis plicatae total flavonoids.
The present invention seeks to be achieved through the following technical solutions:
The application of Radix hemerocalis plicatae in the preparation antidepressant drug.
Application in the 5-hydroxy tryptamine medicine of Radix hemerocalis plicatae in preparation rising brain cortex.
Application in the norepinephrine medicine of Radix hemerocalis plicatae in preparation rising brain cortex.
Application in the dopamine medicine of Radix hemerocalis plicatae in preparation rising brain cortex.
Application in the 5-hydroxyindoleacetic acid medicine of Radix hemerocalis plicatae in preparation rising brain cortex.
The application of Radix hemerocalis plicatae in preparation antagonism reserpine medicine.
The application of Radix hemerocalis plicatae total flavonoids in the preparation antidepressant drug.
Application in the 5-hydroxy tryptamine medicine of Radix hemerocalis plicatae total flavonoids in preparation rising brain cortex.
Application in the norepinephrine medicine of Radix hemerocalis plicatae total flavonoids in preparation rising brain cortex.
Application in the dopamine medicine of Radix hemerocalis plicatae total flavonoids in preparation rising brain cortex.
Application in the 5-hydroxyindoleacetic acid medicine of Radix hemerocalis plicatae total flavonoids in preparation rising brain cortex.
The application of Radix hemerocalis plicatae total flavonoids in preparation antagonism reserpine medicine.
Radix hemerocalis plicatae total flavonoids can be according to the method preparation of prior art, and the present invention provides a kind of method for preparing Radix hemerocalis plicatae total flavonoids simultaneously:
Step 1: Radix hemerocalis plicatae 60-95% alcohol heating reflux is extracted 1-4 time, each 0.5-1 hour, filter, merging filtrate reclaims ethanol, gets extractum A, residue I;
Step 2: residue I 20-80% ethanol heating extraction 1-4 time, each 20-60min filters, and merging filtrate reclaims ethanol, gets extractum B;
Step 3: with extractum A distilled water diluting after-filtration, by macroporous resin column, it is transparent that distilled water is washed till effluent, with 2-6 times of column volume of 20%-95% ethanol elution, reclaims ethanol respectively, obtains sample 1;
Step 4: with extractum B distilled water diluting after-filtration, by macroporous resin column, it is transparent that distilled water is washed till effluent, successively uses 2-6 times of column volume of 20%-95% ethanol elution, reclaims ethanol respectively, obtains sample 2.
Step 5: the sample of blend step 3, step 4 makes Radix hemerocalis plicatae total flavonoids.
The described method for preparing Radix hemerocalis plicatae total flavonoids is preferably:
Step 1: Radix hemerocalis plicatae 95% soak with ethanol is spent the night, 95% ethanol heating extraction 3 times, each 0.5-1 hour, filter, merging filtrate reclaims ethanol, gets extractum A, residue I;
Step 2: residue I is with 60% ethanol heating extraction 2 times, and each 40min filters, and merging filtrate reclaims ethanol, extractum B;
Step 3: with extractum A distilled water diluting after-filtration, by the AB-8 macroporous resin column of having handled well, it is transparent that distilled water is washed till effluent, with 3 times of column volumes of 60% ethanol elution, reclaims ethanol, obtains sample 1;
Step 4: with extractum B distilled water diluting after-filtration, by the AB-8 macroporous resin column of having handled well, it is transparent that distilled water is washed till effluent, with 3 times of column volumes of 60% ethanol elution, reclaims ethanol, obtains sample 2;
Radix hemerocalis plicatae total flavonoids of the present invention can also be made the acceptable any conventional dosage form of pharmaceutics, for example preparations such as capsule, tablet, granule, powder, oral liquid, pill by preparation process routinely.For above-mentioned dosage form can be realized, need when these dosage forms of preparation, to add the pharmacy acceptable auxiliary, for example: filler, disintegrating agent, lubricant, suspending agent, binding agent, sweeting agent, correctives, antiseptic, substrate etc.Filler comprises: starch, pregelatinized Starch, lactose, mannitol, chitin, microcrystalline Cellulose, sucrose etc.; Disintegrating agent comprises: starch, pregelatinized Starch, microcrystalline Cellulose, carboxymethyl starch sodium, crospolyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose etc.; Lubricant comprises: magnesium stearate, sodium lauryl sulphate, Pulvis Talci, silicon dioxide etc.; Suspending agent comprises: polyvinylpyrrolidone, microcrystalline Cellulose, sucrose, agar, hydroxypropyl emthylcellulose etc.; Binding agent comprises, starch slurry, polyvinylpyrrolidone, hydroxypropyl emthylcellulose etc.; Sweeting agent comprises: saccharin sodium, Aspartane, sucrose, cyclamate, enoxolone etc.; Correctives comprises: sweeting agent and various essence; Antiseptic comprises: parabens, benzoic acid, sodium benzoate, sorbic acid and its esters, benzalkonium bromide, fixed, the Folium eucalypti globueli (Eucalyptus globulus Labill.) wet goods of acetic acid chloroethene; Substrate comprises: PEG6000, PEG4000, insect wax etc.
Has antidepressant effect through pharmacological testing proof Radix hemerocalis plicatae of the present invention and Radix hemerocalis plicatae total flavonoids, can significantly improve the 5-HT of rat brain cortex, NE, the 5-HT of DA content and Hippocampus, NE content, with model group relatively there were significant differences (P＜0.05-0.01), after the administration 28 days, compare with normal group, the Open-Field method level of model group rat is passed through the lattice number, erectting number of times obviously reduces, difference has significance (P＜0.01), compare with model group, Radix hemerocalis plicatae or Radix hemerocalis plicatae total flavonoids capsule are big, in, the Open-Field method level of small dose group and fluoxetine Hydrochloride group rat is passed through the lattice number, erectting time number average has increase, has significant difference (P＜0.05-0.01); After the administration 28 days, compare with model group, Radix hemerocalis plicatae or the large, medium and small dosage group of Radix hemerocalis plicatae total flavonoids capsule rat sucrose solution consumption increase, and difference has significance (P＜0.05-0.01); Compare with model group, the dead time during the mandatory swimming of Radix hemerocalis plicatae or the large, medium and small dosage group of Radix hemerocalis plicatae total flavonoids capsule rat significantly reduces, and has significant difference (P＜0.01).Body temperature decline effect due to the remarkable antagonism reserpine of the large, medium and small dosage energy of Radix hemerocalis plicatae or Radix hemerocalis plicatae total flavonoids capsule and mice blepharoptosis and akinetic effect (P＜0.05-0.01).Following experimental example further specifies the present invention by Radix hemerocalis plicatae according to the mind calming resolving depression capsule pharmacological testing of the method for embodiment 1 preparation.
Experimental example 1 mind calming resolving depression capsule is to the therapeutical effect of rat chronic stress depression disease
Test objective
Adopt and chronicly comprehensively stress method set up depression rat model, observe the capsular antidepressant effect of Chinese medicine mind calming resolving depression.
Be subjected to the reagent thing
Mind calming resolving depression capsule dry powder, lot number:, the Beijing University Of Chinese Medicine Traditional Chinese Medicine College provides.Fluoxetine Hydrochloride, batch number: A126471, lot number of the repackaged products: 051020, factory: U.S. Eli Lilly and Company Limited, packing factory: gift lysol continent pharmaceutical Co. Ltd.Norepinephrine (NE), dopamine (DA), 5-hydroxy tryptamine (5-HT), 3,4-dihydroxyphenyl acetic acid (DOPAC), 4-hydroxy-3-methoxy-.alpha.-toluic acid. (HVA), 5-hydroxyindoleacetic acid (5-HIAA), 3,4-dihydroxy Bian amine (DHBA) more than is SIGMA company product.Di-n-butylamine (the import packing of chemical reagent purchasing station, Shanghai), B-8 ion-pairing agent (Tianjin chemical reagent two factories), methanol (top grade is pure, the Beijing Chemical Plant), other reagent are homemade analytical pure.
Date processing
All experimental datas are represented with x ± s.Continuous data carries out check between variance analysis and group by the SPSS10.0 statistical software.
Laboratory animal
SPF level Sprague-Dawley (SD) rat, body weight 250-300g, male and female half and half, animal licence numbering: SCXK (capital) 2002-0003, Beijing Vital River Experimental Animals Technology Co., Ltd. provides.
The raising condition
After animal entered Animal Lab., the every cage of rat was put 5 in a suitable place to breed, raised and enabled after 3 days; The every cage of normal rats is put 5 in a suitable place to breed, and all single cage of the rat of dosage group, mind calming resolving depression capsule low dose group, fluoxetine Hydrochloride group is raised in model group, mind calming resolving depression capsule in high dose group, the mind calming resolving depression capsule.Animal housing's illumination abundance, heating ventilation and air-conditioning equipment is good, and room temperature is controlled at 23-25 ℃, and relative humidity is 40-70%.Laboratory is sterilization regularly routinely.
The foundation of chronic stress depression model
Rat is placed under the circadian rhythm illumination condition, and the ad lib water inlet is raised 3 days to conform, and touches animal every day to adapt to experimenter's operation.After conforming, rat is divided into blank group (hereinafter to be referred as the blank group), depression model group (hereinafter to be referred as model group) at random.Model group is raised according to all single cage of document, stimulation with chronic Unpredictability, cooperate lonely supporting, various stimulating factors are used in 28d by random method, comprise (4 ℃ of frozen water swimming, 5min), (45 ℃ of thermostimulations, 5min), rock (frequency 160 times/minute, 3min), folder tail (1min), prohibit water (24h), (24h) put upside down in fasting (24h) round the clock, give a kind of stimulation every day, every kind stimulates accumulative total to use 4 times, and order makes animal can not expect the generation that stimulates at random, after giving stimulation in 28 days, carrying out OpenField behavioristics measures, experiment of rat forced swimming and sucrose solution consumption experiment, whether successful with the chronic stress depression model of evaluation model group rat, the rat of modeling success is divided into model group at random, mind calming resolving depression capsule in high dose group, dosage group in the mind calming resolving depression capsule, mind calming resolving depression capsule low dose group, the fluoxetine Hydrochloride group.Make Open Field behavioristics, the experiment of rat forced swimming of respectively organizing rat roughly consistent as far as possible, as the index before the administration with the experimental result of sucrose solution consumption experiment.After the beginning administration, accepted once different stress stimulations again every 2 days, stimulation mode and degree are identical during with modeling, until the experiment end.
Medication
After giving stimulation in 28 days, be divided into dosage group, mind calming resolving depression capsule low dose group, fluoxetine Hydrochloride group in model group, mind calming resolving depression capsule in high dose group, the mind calming resolving depression capsule at random.Can calculate the dose,equivalent of each animal according to " laboratory animal with the people by body surface area than equivalent conversion ratio ", as middle dosage.The capsular heavy dose of mind calming resolving depression is the 122.31mg/kg body weight, and middle dosage is the 40.77mg/kg body weight, and low dosage dosage is the 13.59mg/kg body weight; The Western medicine group gives fluoxetine Hydrochloride, and dosage is the 3.6mg/kg body weight, and all by the concentration gastric infusion of 1ml/100g body weight, model group gives normal saline 1ml/100g body weight.Irritated stomach altogether 28 days.
Experimental index is observed
1, Open-Field behavioristics measures
In the variation that gives to stimulate afterwards and respectively organized with the observation of Open-field method in 28 days after the administration in 28 days rat behavior.According to document self-control Open Field inspection box, diameter 90cm, high 40cm, perisporium are black, and the bottom surface is divided into 25 lattice that area equates by fan-shaped.Passing through bottom surface grid number with rat is horizontal score, and it is 1 minute that animal is passed through 1 lattice (striding into more than 3 pawls); With rat biped built on stilts number of times is vertical score, and the animal biped leaves the bottom surface and is sign, no matter animal how long stand be 1 minute until putting down biped.Every animal is once measured at every turn, and the time is 5min.Utilize camera head to write down the variation of its behavior, analysis result.
2, rat forced swimming experiment
After giving to stimulate back and administration in 28 days after 28 days, according to document rat is put into internal diameter 20cm, in the glass circle cylinder of high 40cm, depth of water 24cm in 1 in the every cylinder, cylinder, water temperature 28-29 ℃, (every cylinder water is only used 1 time) observes 5min continuously, and the accumulative total rat stops to struggle in water, be floating erectility, it is the dead time with the persistent period that keeps head to keep afloat that limb motion is once in a while only arranged.(this experiment the previous day, every rat was swum 15min in advance, took out to be put in to dry under the lamp subsequently and put into cage.)
3, sucrose solution consumption experiment
After 28 days, carry out sucrose solution consumption experiment after giving stimulation in 28 days and after the administration.Every rat adds 1% sucrose solution 100ml when carrying out sucrose solution consumption experiment, and all rats carry out fasting simultaneously, calculates the amount that rat 1h drinks 1% sucrose solution.
4, the mensuration of cortex and Hippocampus monoamine neurotransmitter
Method of drawing material: behind the sacrificed by decapitation animal, separate full brain, isolate prefrontal cortex and Hippocampus on ice rapidly, after the frozen Guan Zhongyong liquid nitrogen quick freezing of the rearmounted 1.5ml of title quality, put into-80 ℃ of refrigerators and be saved to mensuration.
The processing of sample: the solution that add different volumes according to cerebral tissue quality difference: every 100mg cerebral tissue adds 1ml organized processing liquid (the DHBA 25 μ l of 0.15mmol/L HClO4 and 2ng/ μ l, ultrasonic homogenate 30s under the condition of ice bath, then at 15000 rev/mins, 4 ℃ of centrifugal 20min, supernatant are used for neurotransmitter and measure.
Neurotransmitter detects: adopt highly effective liquid phase chromatographic system (HPLC)-electrochemical detector (ECD) to detect the concentration of monoamine neurotransmitter and main metabolites thereof, comprise NE, DOPAC, DA, 5-HIAA, HVA, 5-HT.Get supernatant 10 μ l sample introductions and measure, each neurotransmitter content is with the wet Mass Calculation of ng/g tissue.Chromatographic condition: chromatographic condition: Nova-pak C18 chromatographic column (4mm * 150mm, 5 μ m); Mobile phase 50mmolL
-1Citric acid-sodium-acetate buffer pH 3.5 (includes 1.0mmolL
-1The B-8 ion-pairing agent, 1.8mmolL
-1Di-n-butylamine, 0.3mmolL
-1The EDTA disodium, 4% methanol), flow velocity 1.0mLmin
- 1, the vitreous carbon working electrode, detection cell voltage is+0.75V; 3,4-dihydroxy benzylamine DHBA is an internal standard substance, and each key component is carried out quantitatively with internal standard method in the sample.
Experimental result
1. mind calming resolving depression capsule is to the influence of 5-HT, NE, DA, 5-HIAA, HVA and the DOPAC content of rat brain cortex
The 5-HT of model group rat brain cortex, NE, DA, 5-HIAA and DOPAC content and normal group significantly reduce (P＜0.01); The 5-HT of the heavy dose of group of mind calming resolving depression capsule rat brain cortex, NE, DA and 5-HI are from (the P＜0.05-0.01) that significantly raises with model group comparison content, the DOPAC content and the model group of the heavy dose of group of mind calming resolving depression capsule rat brain cortex relatively have rising trend, but difference does not have significance (P＞0.05), the 5-HT of dosage group rat brain cortex and NE and model group comparison content significantly raise (P＜0.01) in the mind calming resolving depression capsule, the DA of dosage group rat brain cortex in the mind calming resolving depression capsule, DOPAC and 5-HIAA content and model group relatively have rising trend, but difference does not have significance (P＞0.05), the 5-HT of mind calming resolving depression capsule small dose group rat brain cortex, NE and 5-HIAA content and model group relatively have rising trend, but difference does not have significance (P＞0.05), sees Table 1-3.
Table 1 mind calming resolving depression capsule to the influence of depressed rat brain cortex 5-HT and 5-HIAA content (ng/g,
)
Annotate: compare * P＜0.01 with model group; * P＜0.05
Table 2 mind calming resolving depression capsule to the influence of depressed rat brain cortex DA and DOPAC content (ng/g,
)
Annotate: compare * P＜0.01 with model group; * P＜0.05
Table 3 mind calming resolving depression capsule to the influence of depressed rat brain cortex NE and HVA content (ng/g,
)
Annotate: compare * P＜0.01 with model group; * P＜0.05
2. mind calming resolving depression capsule is to the influence of 5-HT, NE, DA, 5-HIAA, HVA and the DOPAC content of rat hippocampus
The 5-HT of model group rat hippocampus, NE and DA content and normal group significantly reduce (P＜0.01), and its 5-HI relatively has reduction trend from, DOPAC and HVA content and normal group, but difference does not have significance (P＞0.05); The 5-HT of the big or middle dosage group of mind calming resolving depression capsule rat hippocampus and NE and the model group comparison content (P＜0.05-0.01) that significantly raises, the 5-HT of mind calming resolving depression capsule small dose group rat hippocampus and NE content and model group relatively have rising trend, but difference does not have significance (P＞0.05), sees Table 4-6.
Table 4 mind calming resolving depression capsule to the influence of depressed rat hippocampus 5-HT and 5-HIAA content (ng/g,
)
Annotate: compare * P＜0.01 with model group; * P＜0.05
Table 5 mind calming resolving depression capsule to the influence of depressed rat hippocampus DA and DOPAC content (ng/g,
)
Annotate: compare * P＜0.01 with model group; * P＜0.05
Table 6 mind calming resolving depression capsule to the influence of depressed rat hippocampus NE and HVA content (ng/g,
)
Annotate: compare * P＜0.01 with model group; * P＜0.05
3. mind calming resolving depression capsule is to the influence of rat Open-field behavior
Before the administration, the Open-Field method level of model group, the large, medium and small dosage group of mind calming resolving depression capsule and fluoxetine Hydrochloride group rat is passed through the lattice number, is erect number of times there was no significant difference (P＜0.05), but with normal group significant difference (P＜0.01) is arranged more all; After the administration 28 days, compare with normal group, the Open-Field method level of model group rat is passed through the lattice number, erectting number of times obviously reduces, difference has significance (P＜0.01), compare with model group, mind calming resolving depression capsule is big, in, the Open-Field method level of small dose group and fluoxetine Hydrochloride group rat is passed through the lattice number, erect time number average increase is arranged, has significant difference (P＜0.05-0.01), with before on the same group the administration relatively, mind calming resolving depression capsule is big, the Open-Field method level of middle dosage group and fluoxetine Hydrochloride group rat is passed through the lattice number, erectting number of times obviously increases, and has significant difference (P＜0.05-0.01).See Table 7.
Table 7 mind calming resolving depression capsule is to the influence of depressed rat Open-field behavior
Annotate: compare Δ P＜0.01 with normal group
Compare * P＜0.01 with model group; * P＜0.05
With #P＜0.01 relatively before on the same group the administration; ##P＜0.05
4. mind calming resolving depression capsule is to the influence of rat sucrose solution consumption
Before the administration, model group, mind calming resolving depression capsule is big, in, small dose group and fluoxetine Hydrochloride group rat sucrose solution consumption comparing difference do not have significance (P＜0.05), but significant difference (P＜0.01) is more all arranged with normal group, after the administration 28 days, compare with normal group, model group rat sucrose solution consumption obviously reduces, difference has significance (P＜0.01), compare with model group, mind calming resolving depression capsule is big, in, small dose group and fluoxetine Hydrochloride group rat sucrose solution consumption increase, difference has significance (P＜0.05-0.01), with before on the same group the administration relatively, mind calming resolving depression capsule is big, middle dosage group and fluoxetine Hydrochloride group rat sucrose solution consumption increase, and difference has significance (P＜0.01, P＜0.05).See Table 8.
Table 8 mind calming resolving depression capsule to the influence of depressed rat sucrose solution consumption (ml,
)
Annotate: compare Δ P＜0.01 with normal group
Compare * P＜0.01 with model group; * P＜0.05
With #P＜0.01 relatively before on the same group the administration; ##P＜0.05
5. mind calming resolving depression capsule is to the influence of the mandatory swimming swimming of rat
Before the administration, model group, the large, medium and small dosage group of mind calming resolving depression capsule and the dead time of fluoxetine Hydrochloride group rat during mandatory swimming, there was no significant difference (P＞0.05), but more all be significantly increased with normal group all has significant difference (P＜0.05-0.01); After the administration 28 days, compare with normal group, the dead time of model group rat during mandatory swimming significantly increases, has significant difference (P＜0.01), compare with model group, large, medium and small dosage group of mind calming resolving depression capsule and the dead time of fluoxetine Hydrochloride group rat during mandatory swimming significantly reduce, has significant difference (P＜0.01), with before on the same group the administration relatively, the big or middle dosage group of mind calming resolving depression capsule and fluoxetine Hydrochloride group rat sucrose solution consumption increase, and difference has significance (P＜0.05-0.01).See Table.
The influence that table 9 mind calming resolving depression capsule is swum to the mandatory swimming of depressed rat (s,
)
Annotate: compare Δ P＜0.01 with normal group; Δ Δ P＜0.05
Compare * P＜0.01 with model group; * P＜0.05
With #P＜0.01 relatively before on the same group the administration; ##P＜0.05
Experimental example 2 mind calming resolving depression capsules are to the influence of reserpine antagonistic experiment
Test objective
Adopt the capsular antidepressant effect of reserpine antagonistic experiment research mind calming resolving depression.
Medicine and preparation
Mind calming resolving depression capsule dry powder, the Beijing University Of Chinese Medicine Traditional Chinese Medicine College provides.Fluoxetine Hydrochloride, batch number: A126471, lot number of the repackaged products: 051020, factory: U.S. Eli Lilly and Company Limited, packing factory: gift lysol continent pharmaceutical Co. Ltd.The reserpine injection, ShangHai Fudan Fuhua Pharmaceutical Co., Ltd, lot number:
050601，
Date processing
Experimental data is all represented with x ± s, relatively t check between date processing employing group.Open eyes and can not and move and relatively adopt chi-square criterion between the group that can not observe
Laboratory animal
SPF level ICR, male and female half and half, mice, 18～20g, Beijing dimension tonneau China Experimental Animal Center provides, the quality certification number: SCXK (capital) 2002-2003.
Experimental technique
1. grouping and administration
Animal is divided into normal group, positive drug group, treatment group at random.Normal group is given normal saline; The positive drug group: the reserpine antagonistic experiment gives fluoxetine (5.2mg/kg), and the treatment component is the large, medium and small dosage group of mind calming resolving depression capsule (is respectively 176.67,58.89,19.63mg/kg).Every group of 20 mices, gastric infusion, 2 1d, 7d continuously.
2. antagonism reserpine experiment
Behind the last administration 1h, lumbar injection reserpine 2.5mg/kg carries out the observation of following experiment and index respectively.
2.1 the observation of anus temperature: behind lumbar injection reserpine 3, the 5h, thermometer probe is inserted the about 1cm of anus measure the anus temperature, data respectively organized in record.
2.2 open eyes can not observation: behind the lumbar injection reserpine 1h, observe the number that mice can not open eyes, calculate percentage rate.
2.3 akinetic observation: behind the lumbar injection reserpine 1h, it is that the circle of 7.5cm is observed 15s that mice is put into diameter, calculates and respectively organizes the mice rate that goes too far.
The result
1. mind calming resolving depression capsule is to the influence of reserpine induced mice body temperature decline
Show as body temperature behind the model group subcutaneous injection of reserpine and descend, significant difference is more all arranged with the normal control group
(P＜0.01); Mouse temperature due to the remarkable antagonism reserpine of the large, medium and small dosage energy of mind calming resolving depression Capsules group descends
(P＜0.05) the results are shown in Table 1.
Table 1 mind calming resolving depression capsule to the influence of the depressed rat temperature of reserpine (℃,
)
Annotate: compare Δ P＜0.01 with normal group;
Compare * P＜0.01, * * P＜0.05 with model group
2. mind calming resolving depression capsule is to the antagonism of reserpine induced mice blepharoptosis
Show as tangible blepharoptosis behind the model group subcutaneous injection of reserpine, can not open eyes, significant difference is more all arranged with the normal control group; Blepharoptosis (P＜0.05-), the results are shown in Table 2 due to the remarkable antagonism reserpine of the large, medium and small dosage energy of mind calming resolving depression Capsules group.
Table 2 mind calming resolving depression capsule is to the antagonism of reserpine induced mice blepharoptosis
Annotate: compare * P＜0.01, * * P＜0.05 with model group
3. mind calming resolving depression capsule is to the akinetic antagonism of reserpine induced mice
Show as behind the model group subcutaneous injection of reserpine motion can not, the rate that goes too far significantly reduces, and with the normal control group significant difference (P＜0.01) is arranged more all; The large, medium and small dosage of mind calming resolving depression Capsules group significantly the motion due to the antagonism reserpine can not, the rate that goes too far significantly high with model group (P＜0.05), the results are shown in Table 3.
Table 3 mind calming resolving depression capsule is to the akinetic antagonism of reserpine induced mice
Annotate: compare * P＜0.01 with model group
Experimental example 3 content of total flavone are measured
The sample of getting among the embodiment 1 and 2 carries out determination of total flavonoids.
Assay method: precision takes by weighing the rutin 0.0101g of drying under reduced pressure to constant weight, and methanol constant volume shakes up in the 50ml volumetric flask, accurately measures 0.0,1.0,2.0,3.0,4.0, and 5.0ml adds 5%NaNO in the 25ml volumetric flask
36ml, mixing is placed 6mi n, adds 10%Al (NO
3)
30.6ml mixing is placed 6min, adds 4%NaOH solution 5ml, to scale, places 15min with 60% ethanol dilution, by the pharmacopeia spectrophotography, is blank with first pipe, surveys trap at 500nm wavelength place.With the trap is abscissa, and concentration is vertical coordinate, the drawing standard curve.
| Concentration (mg/ml) | 0.008 | 0.016 | 0.024 | 0.032 | 0.04 |
| Trap | 0.088798 | 0.20217 | 0.30284 | 0.39565 | 0.48809 |
Regression equation is y=12.401x-0.0021 r=0.9990
Accurately take by weighing sample 1 and 2 each 0.0025g, place the 50ml volumetric flask, measure trap with method and be respectively 0.11867 and 0.12219, be respectively 0.009739 and 0.010022mg/ml by the standard curve calculating concentration.General flavone content is respectively 19.478% and 20.044%.
Following embodiment all can realize the described effect of above-mentioned experimental example
The specific embodiment
Embodiment 1: capsule
The 9900g dry day-lily buds is cut into the segment of 2-3 centimeter length, and 95% soak with ethanol is spent the night.95% ethanol heating extraction 3 times picks up counting from boiling, and the time is respectively 40min, 30min, and 30min filters, and merging filtrate reclaims ethanol, gets extractum A.Residue is with 60% ethanol heating extraction 2 times, and each 40min filters, and merging filtrate reclaims ethanol, extractum B.With extractum A distilled water diluting after-filtration, by the AB-8 macroporous resin column of having handled well, it is transparent that distilled water is washed till effluent, with 3 times of column volumes of 60% ethanol elution, reclaims ethanol, obtains sample 1.Handle extractum B with method, get sample 2.Sample 1 is 64.3101g, and sample 2 is 60.9338g.Paste-forming rate is got dried extractum greater than 1.265%, and by 1: 0-5 adds lactose (or starch or dextrin), mix homogeneously, make granule or direct canned in capsule promptly.
Effect: resolving depression, mind calming is calmed the nerves
Cure mainly: be used for the depressive state card and belong to the motive thanks to void, malaise person.Disease is seen depressed emotion, sighing frequently, and insomnia and dreamful sleep, dream is many easily wakes up, and malaise, susceptible to fear are easily frightened, pale tongue, deep-thready pulse
Total amount symphysis every day medicine 15 grams (each 5 grams), every day 3 times.1 month was 1 course of treatment, with 3 courses of treatment.
Embodiment 2: the preparation of total flavones and check in the Radix hemerocalis plicatae
The 9900g dry day-lily buds is cut into the segment of 2-3 centimeter length, and 95% soak with ethanol is spent the night.95% ethanol heating extraction 3 times picks up counting from boiling, and the time is respectively 40min, 30min, and 30min filters, and merging filtrate reclaims ethanol, gets extractum A.Residue is with 60% ethanol heating extraction 2 times, and each 40min filters, and merging filtrate reclaims ethanol, extractum B.With extractum A distilled water diluting after-filtration, by the AB-8 macroporous resin column of having handled well,, it is transparent that distilled water is washed till effluent, with 3 times of column volumes of 60% ethanol elution, reclaims ethanol, obtains sample 1.Handle extractum B with method, get sample 2.Sample 1 is 64.3101g, and sample 2 is 60.9338g.Paste-forming rate is greater than 1.265%, and aggregate sample 1,2 promptly gets total flavones in the Radix hemerocalis plicatae.
Adopt dissolve with ethanol to carry out following experiment in above-mentioned sample, proved to belong to flavone compound according to the general inspection method of flavone compound.
1, inspected yellow-green fluorescence under the ultra-violet lamp;
2, add and drip concentrated hydrochloric acid 2-3 after a little magnesium powder jolting and drip solution, be pink after 1-2 minute;
3, after the alcoholic solution of sample and 1% aluminum chloride alcoholic solution react by paper speck, put apparent yellow-green fluorescence under the uviol lamp;
4, dropping sodium solution is pale brown color
Embodiment 3: granule
The 9900g dry day-lily buds is cut into the segment of 2-3 centimeter length, and 95% soak with ethanol is spent the night.95% ethanol heating extraction 3 times picks up counting from boiling, and the time is respectively 40min, 30min, and 30min filters, and merging filtrate reclaims ethanol, gets extractum A.Residue is with 60% ethanol heating extraction 2 times, and each 40min filters, and merging filtrate reclaims ethanol, extractum B.With extractum A distilled water diluting after-filtration, by the AB-8 macroporous resin column of having handled well,, it is transparent that distilled water is washed till effluent, with 3 times of column volumes of 60% ethanol elution, reclaims ethanol, obtains sample 1.Handle extractum B with method, get sample 2.Sample 1 is 64.3101g, and sample 2 is 60.9338g.Paste-forming rate is greater than 1.265%, and aggregate sample 1,2 adds dextrin and makes granule through conventional technology.
Embodiment 4: tablet
The 9900g dry day-lily buds is cut into the segment of 2-3 centimeter length, and 95% soak with ethanol is spent the night.95% ethanol heating extraction 3 times picks up counting from boiling, and the time is respectively 40min, 30min, and 30min filters, and merging filtrate reclaims ethanol, gets extractum A.Residue is with 60% ethanol heating extraction 2 times, and each 40min filters, and merging filtrate reclaims ethanol, extractum B.With extractum A distilled water diluting after-filtration, by the AB-8 macroporous resin column of having handled well,, it is transparent that distilled water is washed till effluent, uses 60% ethanol elution, reclaims ethanol, obtains sample 1.Handle extractum B with method, get sample 2.Sample 1 is 64.3101g, and sample 2 is 60.9338g.Paste-forming rate is greater than 1.265%, and aggregate sample 1,2 adds starch, lactose, dextrin after conventional technology is made granule, adds lubricant routinely, is pressed into tablet.
Claims (10)
1. the application of Radix hemerocalis plicatae in preparation antagonism reserpine medicine.
2. the application of Radix hemerocalis plicatae total flavonoids in the preparation antidepressant drug.
3. application as claimed in claim 2 is characterized in that the application in the 5-hydroxy tryptamine medicine of Radix hemerocalis plicatae total flavonoids in preparation rising brain cortex.
4. application as claimed in claim 2 is characterized in that the application in the norepinephrine medicine of Radix hemerocalis plicatae total flavonoids in preparation rising brain cortex.
5. application as claimed in claim 2 is characterized in that the application in the dopamine medicine of Radix hemerocalis plicatae total flavonoids in preparation rising brain cortex.
6. application as claimed in claim 2 is characterized in that the application in the 5-hydroxyindoleacetic acid medicine of Radix hemerocalis plicatae total flavonoids in preparation rising brain cortex.
7. the application of Radix hemerocalis plicatae total flavonoids in preparation antagonism reserpine medicine.
8. the Radix hemerocalis plicatae total flavonoids with antidepressant effect is characterized in that the preparation method of total flavones comprises the steps:
Step 1: Radix hemerocalis plicatae 60-95% alcohol heating reflux is extracted 1-4 time, each 0.5-1 hour, filter, merging filtrate reclaims ethanol, gets extractum A, residue I;
Step 2: residue I 20-80% ethanol heating extraction 1-4 time, each 20-60min filters, and merging filtrate reclaims ethanol, gets extractum B;
Step 3: with extractum A distilled water diluting after-filtration, by macroporous resin column, it is transparent that distilled water is washed till effluent, with 2-6 times of column volume of 20%-95% ethanol elution, reclaims ethanol respectively, obtains sample 1;
Step 4: with extractum B distilled water diluting after-filtration, by macroporous resin column, it is transparent that distilled water is washed till effluent, successively uses 2-6 times of column volume of 20%-95% ethanol elution, reclaims ethanol respectively, obtains sample 2;
Step 5: the sample 1 of blend step 3, the sample 2 of step 4 make Radix hemerocalis plicatae total flavonoids.
9. Radix hemerocalis plicatae total flavonoids as claimed in claim 8 is characterized in that the preparation method of this total flavones comprises the steps:
Step 1: Radix hemerocalis plicatae 95% soak with ethanol is spent the night, 95% ethanol heating extraction 3 times, each 0.5-1 hour, filter, merging filtrate reclaims ethanol, gets extractum A, residue I;
Step 2: residue I is with 60% ethanol heating extraction 2 times, and each 40min filters, and merging filtrate reclaims ethanol, extractum B, residual II;
Step 3: with extractum A distilled water diluting after-filtration, by the AB-8 macroporous resin column of having handled well, it is transparent that distilled water is washed till effluent, with 3 times of column volumes of 60% eluting, reclaims ethanol, obtains sample 1;
Step 4: with extractum B distilled water diluting after-filtration, by the AB-8 macroporous resin column of having handled well, it is transparent that distilled water is washed till effluent, with 3 times of column volumes of 60% eluting, reclaims ethanol, obtains sample 2.
10. a method for preparing Radix hemerocalis plicatae total flavonoids comprises the steps:
Step 1: Radix hemerocalis plicatae 60-95% alcohol heating reflux is extracted 1-4 time, each 0.5-1 hour, filter, merging filtrate reclaims ethanol, gets extractum A, residue I;
Step 2: residue I 20-80% ethanol heating extraction 1-4 time, each 20-60min filters, and merging filtrate reclaims ethanol, gets extractum B;
Step 3: with extractum A distilled water diluting after-filtration, by macroporous resin column, it is transparent that distilled water is washed till effluent, with 2-6 times of column volume of 20%-95% ethanol elution, reclaims ethanol respectively, obtains sample 1;
Step 4: with extractum B distilled water diluting after-filtration, by macroporous resin column, it is transparent that distilled water is washed till effluent, successively uses 2-6 times of column volume of 20%-95% ethanol elution, reclaims ethanol respectively, obtains sample 2;
Step 5: the sample 1 of blend step 3, the sample 2 of step 4 make Radix hemerocalis plicatae total flavonoids.
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| CN 201110137571 CN102210800B (en) | 2008-07-31 | 2008-07-31 | Application of citron daylily and citron daylily general flavone |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102488819A (en) * | 2011-12-12 | 2012-06-13 | 倪健 | Preparing method for daylily flower extract |
| CN103960656A (en) * | 2014-04-03 | 2014-08-06 | 江苏农林职业技术学院 | Method for extracting flavone from day lily |
| CN105853299A (en) * | 2016-05-24 | 2016-08-17 | 广州丹奇日用化工厂有限公司 | Method of extracting daylily extract from dry daylily and application thereof |
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2008
- 2008-07-31 CN CN 201110137571 patent/CN102210800B/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102488819A (en) * | 2011-12-12 | 2012-06-13 | 倪健 | Preparing method for daylily flower extract |
| CN102488819B (en) * | 2011-12-12 | 2015-01-07 | 倪健 | Preparing method for daylily flower extract |
| CN103960656A (en) * | 2014-04-03 | 2014-08-06 | 江苏农林职业技术学院 | Method for extracting flavone from day lily |
| CN105853299A (en) * | 2016-05-24 | 2016-08-17 | 广州丹奇日用化工厂有限公司 | Method of extracting daylily extract from dry daylily and application thereof |
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