CN104069343A - New application of traditional Chinese medicine (TCM) composition - Google Patents

Abstract

The invention provides a new application of a traditional Chinese medicine (TCM) composition. The TCM composition is applied to preparation of medicines for treating depression. The TCM composition comprises the following main active pharmaceutical ingredients: radix polygonum multiflorum preparata, radix astragali, rhizoma polygonati, epimedium herb, barbary wolfberry fruit and salvia miltiorrhiza. The TCM composition can obviously increase the Open-field scores, weights and thymus and spleen coefficients of CUMS (Chronic unpredictable mild stress) model rats, the contents of 5-HT (5-hydroxytryptamine), NE and DA in sea horses, the total number of neurons in the CA3 zones of the sea horses and the percentages of normal neurons, can obviously reduce the contents of 5-HT, NE, DA, ACTH (Adrenocorticotropic Hormone), CORT (Cortisol) and beta-EP in serums, the adrenal gland coefficient and the thickness of the adrenal cortex and has obvious anti-depression effects.

Description

A kind of new purposes of Chinese medicine composition

Technical field

The present invention relates to a kind of new purposes of Chinese medicine composition, be specifically related to a kind of Chinese medicine composition in the new purposes of preparing aspect Cure of depression, belong to field of medicaments.

Background technology

Depression is a kind of common mental sickness, and its prevalence expects the year two thousand twenty will rise to the 2nd.Opinions vary for its pathogenesis, and scholars think it may is the interactive result of the many factors such as psychosocial factor and various biological modifications.Wherein, psychosocial factor mainly contains nervous work and fast pace life, bad inter personal contact, burst life events strike etc.; Biochemical mechanism aspect mainly comprises monoamine hypothesis, receptors hypothesis, endocrine hypothesis, immunity change etc.When monoamine neurotransmitter hypothesis is thought depression, the content of the monoamine neurotransmitter of maincenter (5-HT, NE, DA) reduces.Endocrine hypothesis relates generally to HPA(HPA) axle, stress time, hpa axis excitement, mainly cause CRH, ACTH(corticotropin releasing hormone, thyroliberin) and GC(glucocorticoid) content increase, and the GC Immunosuppression increasing reaction.

Therefore, exploitation becomes for the medicine of monoamine neurotransmitter the technical problem of needing in the industry solution badly.

Summary of the invention

The present invention adopts the screening of CUMS depression model rat to have the Chinese medicine composition product of good Cure of depression effect.

First object of the present invention is to provide a kind of Chinese medicine composition in the application of preparing in Cure of depression medicine.Second object of the present invention is to provide a kind of Chinese medicine composition of Cure of depression;

The 3rd object of the present invention is to provide the preparation method of this Chinese medicine composition.

The object of the invention is to be achieved through the following technical solutions:

One is treated depressed Chinese medicine composition, and the crude drug of this Chinese medicine composition consists of:

Radix Polygoni Multiflori Preparata 35-65 weight portion, Radix Astragali 15-27 weight portion, Rhizoma Polygonati 15-27 weight portion, Herba Epimedii 10-20 weight portion, Fructus Lycii 10-20 weight portion, Radix Salviae Miltiorrhizae 7-14 weight portion.

Further, the crude drug of this Chinese medicine composition consists of:

Radix Polygoni Multiflori Preparata 50 weight portions, the Radix Astragali 22 weight portions, Rhizoma Polygonati 22 weight portions, Herba Epimedii 15 weight portions, Fructus Lycii 15 weight portions, Radix Salviae Miltiorrhizae 11 weight portions.

Wherein, the dosage form of pharmaceutical composition of the present invention is oral liquid, tablet, capsule, pill, granule or drop pill.

" Radix Polygoni Multiflori Preparata " preparation that Radix Polygoni Multiflori Preparata of the present invention records with reference to 2010 editions " Chinese Pharmacopoeia " Firsts.

The present invention also provides the preparation method of this Chinese medicine composition, comprises the steps:

A, take the raw material of weight proportion:

Radix Polygoni Multiflori Preparata 35-65 weight portion, Radix Astragali 15-27 weight portion, Rhizoma Polygonati 15-27 weight portion, Herba Epimedii 10-20 weight portion, Fructus Lycii 10-20 weight portion, Radix Salviae Miltiorrhizae 7-14 weight portion;

B, get Radix Polygoni Multiflori, Radix Salviae Miltiorrhizae, Fructus Lycii and decoct with water, concentrated, add ethanol precipitate with ethanol, repeated hydrogenation sodium oxide regulates pH value to 8.0, filters, and filtrate is adjusted to pH to 7.0, for subsequent use;

C, get Herba Epimedii, Rhizoma Polygonati decocts with water, collecting decoction, filter, leave standstill, get supernatant concentration, add ethanol precipitate with ethanol, repeated hydrogenation sodium oxide regulate pH value to 8.0, filter, filtrate is adjusted to pH to 7.0, for subsequent use;

D, get the Radix Astragali and decoct with water, filter, concentrated;

E, get medicinal liquid prepared by b, c, d and merge.

In addition, after step e, also comprise, add pharmaceutically acceptable adjuvant or complementary composition to be prepared into pharmaceutically conventional preparation, described preparation is oral liquid, tablet, capsule (comprising soft capsule), pill, granule, drop pill.

Wherein, adopt ethanol precipitation twice in step b, ethanol alcohol precipitation concentration is respectively: 70% ethanol, 80% ethanol.

Wherein, adopt ethanol precipitation twice in step c, ethanol alcohol precipitation concentration is respectively: 65% ethanol, 80% ethanol.

For dosage form of the present invention can be realized, need in the time of these dosage forms of preparation, add the acceptable adjuvant of pharmacy, 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, acetic acid chloroethene are determined, Folium eucalypti globueli (Eucalyptus globulus Labill.) wet goods; Substrate comprises: PEG6000, PEG4000, insect wax etc.For making above-mentioned dosage form can realize pharmacy of Chinese materia medica, need add acceptable other adjuvant of pharmacy (Fan Biting " pharmacy of Chinese materia medica ", the adjuvant that in Shanghai Science Press December in 1997 the 1st edition, each dosage form is recorded) when these dosage forms in preparation.

The invention also discloses this Chinese medicine composition in the application of preparing in Cure of depression medicine.

Further, the monoamine neurotransmitter in brain that it is characterized in that raising; Further, it is characterized in that the raising content of 5-hydroxy tryptamine in Hippocampus and/or dopamine.

Further, it is characterized in that reducing the monoamine neurotransmitter in blood; Further, it is characterized in that reducing the content of NE in blood, dopamine, 5-hydroxy tryptamine.

Further, it is characterized in that reducing the content of serum thyroliberin, corticosterone and/or β endorphins.

Further, it is characterized in that increasing neuron number in brain.

Further, it is characterized in that alleviating adrenal gland's pathology changes.

Further, it is characterized in that reducing the adrenal cortex thickness of model mouse.

Experimentation shows, Chinese medicine composition of the present invention (HLS) can obviously raise Open-field scoring, body weight, Thymus and spleen coefficient, Hippocampus 5-HT, NE, the content of DA and the percentage ratio of Hippocampus CA 3 Region neuron total number and normal neurons (the equal <0.05 of P) of CUMS model mouse, can obviously reduce serum 5-HT, NE, DA, ACTH, CORT, β-EP content, adrenal gland's coefficient and adrenal cortex thickness (the equal <0.05 of P).

Brief description of the drawings

Fig. 1 rat hippocampal CA 3 neuron, HE dyeing, × 400,

The rat hippocampal CA 3 neuron of Fig. 1-1, normal group, HE dyeing, × 400,

The rat hippocampal CA 3 neuron of Fig. 1-2, model group, HE dyeing, × 400,

The rat hippocampal CA 3 neuron of Fig. 1-3, model+Sertraline group, HE dyeing, × 400,

The rat hippocampal CA 3 neuron of Fig. 1-4, model+HLS group, HE dyeing, × 400,

The rat hippocampal CA 3 neuron of Fig. 1-5, model+CF4 group, HE dyeing, × 400

Experimental example 1

1 material

260 of the complete male Kunming mouses of the clean level of 1.1 animals, 6～8 week age, weight 24～29g, Sichuan University's Experimental Animal Center provides, the quality certification: No. 10, the real moving Guan Zhi in river, before experiment, Animal adaptability is raised 3～5 days.Feeding environment, 20～25 DEG C of room temperatures, humidity 55～70%, the drinking-water of freely ingesting.

1.2 medicines and reagent

Control drug sertraline hydrochloride sheet (Sertraline) (110801), Dalian pfizer inc, is mixed with 37.5,62.5% 0.5%CMC-Na suspension before use.

Experimental agents 1 HLS: the oral liquid of preparing for the embodiment of the present invention 1;

Experimental agents 2 CF1: Radix Polygoni Multiflori Preparata 1000g, Fructus Lycii 300g, Rhizoma Polygonati (system) 440g;

Preparation method: get Radix Polygoni Multiflori 1000g, Fructus Lycii 300g, add to boil and carry three times, merging filtrate, filter, heating is concentrated into syrupy shape, lets cool, add ethanol and make to reach 70% containing alcohol amount, leave standstill 12 hours, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, then leaves standstill 24 hours, filters, filtrate regulates pH to 8.0 with 10% NaOH, places 24 hours, filters, filtrate regulates pH to 7.0 with 10% hydrochloric acid, reclaims ethanol, and medicinal liquid is for subsequent use.Get Rhizoma Polygonati 440g, add 10 times of water gagings, boil and carry twice, each 1.5 hours, filter, merging filtrate, leaves standstill, and gets supernatant concentration to appropriate, after letting cool, add an alcohol to reaching 65% containing alcohol amount, leave standstill 24 hours, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leave standstill, 24 hours, to filter, filtrate regulates pH value to 8.0 with 10% sodium hydroxide solution, leave standstill 24 hours, filter, filter, filtrate regulates pH to 7.0 with 10% hydrochloric acid, reclaims ethanol, and medicinal liquid is for subsequent use.Merge above reserve liquid, stir evenly, cold preservation is placed 12 hours, filters, and adds water to 1000ml, filters, and filtrate regulates pH value to 7.0 with 10% sodium hydroxide, and heated and boiled sterilizing, to obtain final product.

Experimental agents 3 CF2: Herba Epimedii 300g

Preparation method: take epimedium herb 300g, add 10 times of water gagings, boil and carry twice, each 1.5 hours, filter, merging filtrate, leave standstill, get supernatant concentration to appropriate, after letting cool, add an alcohol to reaching 65% containing alcohol amount, leave standstill 24 hours, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leave standstill, 24 hours, filter, filtrate regulates pH value to 8.0 with 10% sodium hydroxide solution, leave standstill 24 hours, filter, filter, filtrate regulates pH to 7.0 with 10% hydrochloric acid, reclaim ethanol, filter, add water to 1000ml, filter, filtrate regulates pH value to 7.0 with 10% sodium hydroxide, heated and boiled sterilizing, obtain.

Experimental agents 4 CF3: epimedium herb 300g, Radix Polygoni Multiflori Preparata 1000g;

Preparation method: take epimedium herb 300g, add 10 times of water gagings, boil and carry twice, each 1.5 hours, filter, merging filtrate, leaves standstill, and gets supernatant concentration to appropriate, after letting cool, add an alcohol to reaching 65% containing alcohol amount, leave standstill 24 hours, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leave standstill, 24 hours, to filter, filtrate regulates pH value to 8.0 with 10% sodium hydroxide solution, leave standstill 24 hours, filter, filter, filtrate regulates pH to 7.0 with 10% hydrochloric acid, reclaims ethanol, for subsequent use; Take 1000g Radix Polygoni Multiflori Preparata, add to boil and carry three times, merging filtrate, filter, it is strong that heating is concentrated into syrup, lets cool, add ethanol and make to reach 70% containing alcohol amount, leave standstill 12 hours, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, then leaves standstill 24 hours, filters, filtrate regulates pH to 8.0 with 10% NaOH, places 24 hours, filters, filtrate regulates pH to 7.0 with 10% hydrochloric acid, reclaims ethanol, for subsequent use; Get Herba Epimedii extracting solution, Radix Polygoni Multiflori extracting solution stirs evenly, cold preservation is placed 12 hours, filters, and adds water to 1000ml, filters, filtrate is with 10% sodium hydroxide adjusting pH value to 7.0, heated and boiled sterilizing, to obtain final product.

Experimental agents 5 CF4: Herba Epimedii 300g, Radix Astragali 440g, Radix Polygoni Multiflori Preparata 1000g;

Take epimedium herb 300g, add 10 times of water gagings, boil and carry twice, each 1.5 hours, filter, merging filtrate, leaves standstill, and gets supernatant concentration to appropriate, after letting cool, add ethanol to reaching 65% containing alcohol amount, leave standstill 24 hours, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leave standstill, 24 hours, filter, filtrate regulates pH value to 8.0 with 10% sodium hydroxide solution, leaves standstill 24 hours, filters, filtrate regulates pH to 7.0 with 10% hydrochloric acid, reclaims ethanol, for subsequent use; Take Radix Astragali 440g, add 10 times of water gagings and extract three times, each 1.5 hours, merge extractive liquid,, filtered, concentrated for subsequent use; Take 1000g Radix Polygoni Multiflori Preparata, add decocting in water and carry three times, merging filtrate, filter, heating is concentrated into syrupy shape, lets cool, add ethanol and make to reach 70% containing alcohol amount, leave standstill 12 hours, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, then leaves standstill 24 hours, filters, filtrate regulates pH to 8.0 with 10% NaOH, places 24 hours, filters, filtrate regulates pH to 7.0 with 10% hydrochloric acid, reclaims ethanol, for subsequent use; Get Herba Epimedii extracting solution, Radix Astragali extractive solution, Radix Polygoni Multiflori Preparata extracting solution and stir evenly, cold preservation is placed 12 hours, filters, and adds water to 1000ml, filters, and filtrate is with 10% sodium hydroxide adjusting pH value to 7.0, and heated and boiled sterilizing, to obtain final product.

Experimental agents 6 CF5: Radix Polygoni Multiflori Preparata 1000g, Radix Astragali 440g, Herba Epimedii 300g, Radix Salviae Miltiorrhizae 220g;

Preparation method: get Radix Polygoni Multiflori Preparata 1000g, Radix Salviae Miltiorrhizae 220g, adds decocting in water and carry three times, merging filtrate, filter, heating is concentrated into syrupy shape, lets cool, add ethanol and make to reach 70% containing alcohol amount, leave standstill 12 hours, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, then leaves standstill 24 hours, filters, filtrate regulates pH to 8.0 with 10% NaOH, places 24 hours, filters, filtrate regulates pH to 7.0 with 10% hydrochloric acid, reclaims ethanol, and medicinal liquid is for subsequent use.Take epimedium herb 300g, add 10 times of water gagings, boil and carry twice, each 1.5 hours, filter, merging filtrate, leaves standstill, and gets supernatant concentration to appropriate, after letting cool, add an alcohol to reaching 65% containing alcohol amount, leave standstill 24 hours, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leave standstill, 24 hours, to filter, filtrate regulates pH value to 8.0 with 10% sodium hydroxide solution, leave standstill 24 hours, filter, filter, filtrate regulates pH to 7.0 with 10% hydrochloric acid, reclaims ethanol, for subsequent use.Take Radix Astragali 440g, add 10 times of water gagings and extract three times, each 1.5 hours, merge extractive liquid,, filtered, concentrated for subsequent use.Said extracted liquid is mixed and stirred evenly, and cold preservation is placed 12 hours, filters, and adds water to 3000ml, filters, and filtrate regulates pH value to 7.0-7.5 with 10% sodium hydroxide, and heated and boiled sterilizing, to obtain final product.

" Radix Polygoni Multiflori Preparata " used in above-mentioned Experimental agents and " Rhizoma Polygonati (system) " is respectively the processed product of Radix Polygoni Multiflori and Rhizoma Polygonati, the preparation method of " Radix Polygoni Multiflori Preparata " and " Rhizoma Polygonati processed product " that concrete preparation method is recorded with reference to 2010 editions " Chinese Pharmacopoeia " Firsts.

1.3 key instrument MK3 microplate reader (U.S. Thermo); Allegra X-22R Centrifuge(U.S. Backman); 81m-25 optical microscope (Olympus company); ZZ-6 mice autonomic activities instrument (Chengdu TME Technology Co., Ltd.), TH-86-340-WA-80 DEG C of ultra cold storage freezer (Germany), 1/0,000 Sartorius electronic analytical balances (Germany); 1/000 electronic analytical balances (Germany); 250Q fixator of rat; The wooden spacious case of uncovered etc.

2 methods

2.1 experiment

2.1.1 grouping is chosen 96 mices with administration and is divided into 8 groups by body weight stratified random, i.e. normal (0.5%CMC-Na20mlkg-1d-1), positive control (Sertraline, 5mgkg-1d-1), HLS and CF1 thereof, 2,3,4,5(all give corresponding oral liquid 20mlkg-1d-1) totally 8 groups, 12/group, continuous gastric infusion 7d respectively.

2.1.2 outstanding tail is tested 1h after last administration, and mousetail 1-2cm place is suspended on iron stand, with immobilization with adhesive tape, allows mice keep upset hanging posture adaptation 1min, is determined at the dead time in 5min.

2.1.3 swimming test is chosen 96 Kunming mouses, press 2.1.1 step, after last administration 1h, mice is placed in to the cuboid white plastic box of 25 ± 1 DEG C of water temperatures, water capacity 20 × 10 × 13cm3, swimming 6min, is suspended from the water surface dead time after measuring in 5min.

2.2 statistical analysis experimental datas with represent, SPSS17.0 software kit carries out one factor analysis of variance to data, variance test homogeneous, and p>0.05 uses LSD method, and p<0.05 adopts Dunnett T3 method, and p<0.05 is obvious difference.* represent p<0.05 compared with normal group, # represents P<0.05 compared with model group.

3 results

Each Experimental agents the results are shown in Table 1 to the impact of mouse tail suspension and non-swimming time.

The impact of table 1 on mouse tail suspension and non-swimming time

Group	Outstanding tail time (TST)/s	Dead time (FST)/s
			Normal group	138.20±53.46	94.00±54.23
Sertraline	82.60±66.70*	52.73±34.59*
			HLS	86.90±54.30*	59.91±34.77*
CF1	117.78±35.22	98.00±40.81
			CF2	117.54±63.14	80.82±47.16
CF3	107.00±51.15	80.18±35.35
			CF4	81.22±29.73*	64.00±23.50
CF5	121.89±58.28	89.45±42.71

From table 1, with normal group ratio, HLS and CF4 group can significantly shorten the mouse tail suspension dead time (P<0.05), and all the other organize equal no difference of science of statistics (P>0.05); HLS group can significantly shorten mice non-swimming time (P<0.05), and CF4 group has the dead time trend of shortening (P=0.083), and all the other each groups are all without impact (P>0.05).

Experimental example 2

1 material

120 of the complete male Wistar rats of 1.1 animal SPF level, 7～8 week age, weight 200 ± 20g, Da Shuo bio tech ltd, Chengdu provides, the quality certification: SCXK (river) 2008-24, experiment prospective adaptation is raised 1 week.Feeding environment, 20～25 DEG C of room temperatures, humidity 55～70%, the drinking-water of freely ingesting.

1.2 medicines and reagent

Control drug: sertraline hydrochloride sheet (Sertraline) (110801), Dalian pfizer inc, is mixed with 37.5,62.5% 0.5%CMC-Na suspension before use.

Experimental agents 1:HLS, with experimental example 1;

Experimental agents 2:CF4 is that embodiment 1 prepares oral liquid.

Other reagent: autogamy 0.5%CMC-Na liquid; 5-hydroxy tryptamine (5-HT), norepinephrine (NE), dopamine (DA), thyroliberin (ACTH), corticosterone (CORT) and β endorphins (β-EP) Elisa detection kit (201204), by upper Haifeng county, Xiang bio tech ltd provides; Analytical pure PBS, chloral hydrate, paraformaldehyde etc.

1.3 key instrument MK3 microplate reader (U.S. Thermo); Allegra X-22R Centrifuge(U.S. Backman); 81m-25 optical microscope (Olympus company); ZZ-6 mice autonomic activities instrument (Chengdu TME Technology Co., Ltd.), TH-86-340-WA-80 DEG C of ultra cold storage freezer (Germany), 1/0,000 Sartorius electronic analytical balances (Germany); 1/000 electronic analytical balances (Germany); 250Q fixator of rat; The wooden spacious case of uncovered etc.

2 methods

2.1 experiment

2.1.1 grouping and administration

120 of male Wistar rats, are tested and are done behavioristics's scoring by Open-field, and the rat of choosing 30～120 points of scores enters experiment, according to the screening experiment result of mice, by Open-field experiment scoring, adopt randomized block design, be divided into 5 groups, every group more than 16.1. normal group: normally raise; 2. model group: accept lonely the supporting of 21d CUMS+ and prepare depression model; 3. positive drug group: depression model+Sertraline (Sertraline); 4. HLS group: depression model+HLS; 5. CF4 group: depression model+CF4.After modeling 21d, start respectively continuous gastric infusion 21d, and continue modeling.Each group gives corresponding medicine 20mlkg-1d-1 respectively.

2.1.2CUMS+ lonely supporting set up rat depression model

Except the normal raising of normal group (5～6/cage), other adopts lonely supporting, and gives at random two kinds of different stimulations every day, comprising: 1. and cold water swimming (4 DEG C, 5min.Depth of water 10cm, after swimming finishes moves into rat in 25～30 DEG C of heated pools and makes to take out after its limbs improvements); 2. thermostimulation (45 DEG C of hot plate methods, 5min); 3. prohibit water (24h); 4. fasting (24h) (can't help water when fasting, non-fasting while prohibiting water); 5. put upside down round the clock (24h); 6. moist bedding and padding (l0h); 7. constraint 1h; 8. concussion (1min).

2.1.3Open-Field 100cm × 100cm × 50cm wooden case of marking, bottom is divided into 5 × 5 grids, all paints black in wooden case.The environment of keeping quite, is placed in wooden case central authorities by rat, observes the active situation of rat in 3min, records the horizontal score (at least 3 pawls enter grid pass through lattice number) and vertical score (both feet leave bottom surface) summation of rat.

2.1.4 experimental index detects and the general all 1h detection indexs on the same day after medicine of method.

2.1.4.1Open-Field after experiment and body weight administration, measure weekly an Open-field and test score and body weight.

2.1.4.2 Main Organ Coefficients is measured 1h after 21d administration, get 8 Mus (8:00～10:30) for every group, lumbar injection 10% chloral hydrate (0.3mL/100g) anesthesia, femoral artery is got blood and is put to death, on ice platform, break end immediately, get bilateral hippocampus, separately win adrenal gland, thymus, spleen, weigh respectively, calculate organ coefficient.

2.1.4.3 aforementioned the got blood of the assay of Hippocampus and serum 5-HT, NE, DA, leaves standstill about 1h left and right, and the centrifugal 20min of 3000r/min, gets supernatant.Aforementioned got Hippocampus, adds a certain amount of PBS, and homogenate is ditto centrifugal, gets supernatant, and-80 DEG C of freezing preservations are to be measured.5-HT, DA, the assay of NE is undertaken by test kit specification limit.

2.1.4.4 the mensuration of assay ACTH, the CORT of serum ACTH, CORT, β-EP, β-EP is undertaken by test kit specification limit.

2.1.4.5 PATHOMORPHOLOGICAL OBSERVATION OF PULLORUM is respectively organized 8 rats of residue, with 2.5 anesthesia, lies on the back fixing, through the about 100ml of left ventricle saline injection, shreds liver, and 4% the paraformaldehyde perfusion fixation of reinjecting, gets brain, adrenal gland, weighs respectively.Fixing in specimen 4% paraformaldehyde liquid, paraffin embedding, is interrupted section, and every 10 sections are got 1, and each Hippocampus and adrenal gland respectively get 5, HE dyeing.Adopt TSView7 image analysis system (TUCSEN IMAGING TECHNOLOGY, Inc.), under 400 times of light microscopics, neuron sum and normal neurons number in 5 lattices of hippocampal slices CA3 district counting, the normal neurons number ratio of calculating Hippocampus; Under 40 times of light microscopics, measure adrenocortical maximum ga(u)ge.

2.2 statistical analysis experimental datas with represent, SPSS17.0 software kit carries out one factor analysis of variance to data, variance test homogeneous, and p>0.05 uses LSD method, and p<0.05 adopts Dunnett T3 method, and p<0.05 is obvious difference.* represent p<0.05 compared with normal group, # represents P<0.05 compared with model group.

3 results

3.1 impacts on open-field scoring and body weight change

The results are shown in Table 2.Result shows, with normal group comparison, the times of exercise of model group rat reduces (P<0.05); With model group ratio, after administration 1 week, HLS and CF4 obviously do not increase times of exercise (P>0.05), after 2 weeks, there is increase trend, HLS (P=0.064), CF4 (P=0.087), after 3 weeks, HLS is significantly increased times of exercise effect (P<0.05), and CF4 has increase trend (P=0.093).Model group rat body weight compared with normal group significantly reduces (P<0.05); Compared with model group, administration 1,2,3 weeks, HLS and CF4 are all shown in that body weight significantly increases (P<0.05).Illustrate that HLS has obvious treatment, CF4 to have certain improvement effect to depression rat behavioristics.

The impact of table 2 on open-field scoring and body weight change

3.2 impacts on monoamine neurotransmitter in rat blood serum and brain

The results are shown in Table 3.Result shows, with Normal group ratio, model group rat hippocampus monoamine neurotransmitter content reduces (p<0.05), content raise (p<0.05) in blood.With model group comparison, the content of HLS group Hippocampus 5-HT and DA raises (p<0.05), the content rising (p<0.05) of CF4 group Hippocampus DA.In HLS and CF4 group blood, NE, DA, 5-HT all reduce (p<0.05).

The impact of table 3 on monoamine neurotransmitter in rat blood serum and brain

3.3 impacts on rat blood serum ACTH, CORT, β-EP

The results are shown in Table 4.Result shows that the content of model group serum ACTH, CORT, β-EP is all than the remarkable rising of normal group (P<0.05).Through HLS, CF4 treatment, serum ACTH, CORT, β-EP all have obvious reduction (P<0.05).

The impact of table 4 on rat blood serum ACTH, CORT, β-EP

Group	ACTH/ng.L -1	CORT/ng.L -1	β-EP/ng.L -1
				Normal group	42.98±6.72	14.81±4.27	200.94±59.10
Model group	64.29±15.53*	24.54±6.95*	282.93±72.49*
				Model+Sertraline group	47.04±11.34 #	19.77±3.82 #	201.78±34.42 #
Model+HLS group	43.48±11.11 #	17.83±5.51 #	184.23±37.73 #
				Model+CF4 group	49.19±13.99 #	18.36±7.21 #	195.83±67.93 #

3.4 impacts on Rats Organs and Tissues coefficient

The results are shown in Table 5.Result shows, model group than adrenal gland's coefficient of normal rats obviously increase (p<0.05), thymus coefficient, Spleen coefficient reduces (p<0.05).Through treatment, HLS has certain trend (P=0.093) that reduces to adrenal gland's coefficient, thymus coefficient and Spleen coefficient are had to obvious increase effect (p<0.05), CF4 without obvious effect (p>0.05), has increase tendency to thymus coefficient (P=0.053) and Spleen coefficient (P=0.058) to adrenal gland's coefficient.

The impact of table 5 on Rats Organs and Tissues coefficient

Group	Adrenal gland's coefficient	Thymus coefficient	Spleen coefficient
				Normal group	0.0160±0.0021	0.1574±0.0271	0.4256±0.1381
Model group	0.0274±0.0182*	0.1003±0.0466*	0.2096±0.0464*
				Model+Sertraline group	0.0239±0.0071	0.1281±0.0485	0.2466±0.0520
Model+HLS group	0.0212±0.0055	0.1424±0.0458#	0.2914±0.1539#
				Model+CF4 group	0.0232±0.0070	0.1381±0.0743	0.2881±0.1337

3.5 impacts on rat hippocampus, adrenal gland's pathology morphological change

3.5.1HLS with the impact of CF4 on Hippocampus CA 3 Region neuron morphology and number

Normal rats Hippocampus CA 3 Region neuronal cell marshalling, intensive, kernel is high-visible.Compared with normal group, the Hippocampus CA 3 Region neuronal cell of model group rat is arranged sparse, and intercellular substance increases, and kernel disappears, and Cytoplasm pyknosis, is shown in Fig. 1.The shared ratio compared with normal group of the neuron number of model group and normal neurons significantly reduces (P<0.05), compared with model group, the ratio of HLS and CF4 group Hippocampus CA 3 Region neuron number and normal neurons obviously increases (P<0.05), in table 6.

3.5.2HLS with the impact of CF4 on Rat Adrenal Cortex thickness and the change of adrenal gland's pathology

Normal rat adrenal gland clear in structure, ecto-entad is divided into adrenal cortex and medullary substance part, and wherein cortex is divided into again glomerular zone, zona fasciculata and reticular zone; CUMS rat cortex plumpness.The adrenal cortex thickness of model group compared with normal group rat obviously increases (P<0.05), and HLS and CF4 group can obviously reduce the adrenal cortex thickness (P<0.05) of model mouse, in table 6.

The impact of table 6 on rat hippocampus, adrenal gland's pathology morphological change

Group	Neuron number	The ratio (%) of normal neurons	Adrenal cortex thickness (mm)
				Normal group	47.80±4.38	90.31±11.22	793.33±92.45
Model group	33.00±5.52	34.51±19.04*	976.00±105.97*
				Model+Sertraline group	46.25±7.93	77.76±15.48 #	810.12±96.09 #
Model+HLS group	44.80±8.53	73.75±11.77 #	821.11±112.41 #
				Model+CF4 group	43.67±7.84	67.26±14.77 #	841.43±106.54 #

Detailed description of the invention

The preparation of embodiment 1 medicine oral liquid of the present invention

[prescription] Radix Polygoni Multiflori Preparata 1000g, Radix Astragali 440g, Rhizoma Polygonati (system) 440g, Herba Epimedii 300g, Fructus Lycii 300g, Radix Salviae Miltiorrhizae 220g, make 1000ml

[method for making] above Six-element, Radix Polygoni Multiflori Preparata, Radix Salviae Miltiorrhizae, Fructus Lycii decoct with water three times, collecting decoction, filter, filtrate is concentrated into syrupy shape, lets cool, add ethanol and make to reach 70% containing alcohol amount, place, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, places, and filters, regulate pH value to 8.0 with 10% sodium hydroxide solution, place, filter, filtrate regulates pH value to 7.0 with 10% hydrochloric acid solution, reclaims ethanol, and medicinal liquid is for subsequent use; Herba Epimedii, Rhizoma Polygonati decoct with water secondary, and collecting decoction filters, leave standstill, get supernatant concentration to appropriate, let cool, add ethanol to reaching 65% containing alcohol amount, leave standstill, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leaves standstill, and filters, filtrate regulates pH value to 8.0 with 10% sodium hydroxide solution, leaves standstill, and filters, filtrate regulates pH value to 7.0 with 10% hydrochloric acid solution, reclaims ethanol, and medicinal liquid is for subsequent use; The Radix Astragali decocts with water three times, and collecting decoction filters, and filtrate is concentrated into about 95ml, and medicinal liquid is for subsequent use.Merge above medicinal liquid for subsequent use, stir evenly, cold preservation is placed, and filters, and adds water to 1000ml, adds appropriate Sugarless type sweeting agent, filters, and filtrate regulates pH value to 7.0 with 10% sodium hydroxide solution, to obtain final product.

[usage and consumption] is oral, a 10ml, and 1 time on the one, clothes before sleeping, serveing on three months is a course for the treatment of.

[specification] every dress 10ml

[storage] sealing.

The tablet preparation of embodiment 2 medicines of the present invention

Radix Polygoni Multiflori Preparata 1000g, Radix Astragali 440g, Rhizoma Polygonati (system) 440g, Herba Epimedii 300g, Fructus Lycii 300g, Radix Salviae Miltiorrhizae 220g;

Above Six-element, Radix Polygoni Multiflori Preparata, Radix Salviae Miltiorrhizae, Fructus Lycii decoct with water three times, collecting decoction, filter, filtrate is concentrated into syrupy shape, lets cool, add ethanol and make to reach 70% containing alcohol amount, place, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, places, and filters, regulate pH value to 8.0 with sodium hydroxide solution, place, filter, filtrate regulates pH value to 7.0 with 10% hydrochloric acid solution, reclaims ethanol for subsequent use; Herba Epimedii, Rhizoma Polygonati decoct with water secondary, and collecting decoction filters, leave standstill, get supernatant concentration to appropriate, let cool, add ethanol to reaching 65% containing alcohol amount, leave standstill, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leaves standstill, and filters, filtrate regulates pH value to 8.0 with 10% sodium hydroxide, leave standstill, filter, medicinal liquid is for subsequent use; The Radix Astragali decocts with water three times, and collecting decoction filters, and filtrate is concentrated into about 95ml, and medicinal liquid is for subsequent use.Merge above medicinal liquid for subsequent use, stir evenly placement, filter, it is 1.35～1.40(80 DEG C that filtrate is concentrated into relative density) thick paste, add appropriate Icing Sugar, dextrin, mix, granulation, dry, compacting is (coating) in flakes, or above-mentioned thick paste adopts microwave vacuum drying, add appropriate amount of auxiliary materials, compacting is (coating) in flakes, altogether makes 100 doses, to obtain final product.

Capsule (the comprising soft capsule) preparation of embodiment 3 medicines of the present invention

Radix Polygoni Multiflori Preparata 700g, Radix Astragali 300g, Rhizoma Polygonati (system) 300g, Herba Epimedii 200g, Fructus Lycii 200g, Radix Salviae Miltiorrhizae 140g;

Above Six-element, Radix Polygoni Multiflori Preparata, Radix Salviae Miltiorrhizae, Fructus Lycii decoct with water three times, collecting decoction, filter, filtrate is concentrated into syrupy shape, lets cool, add ethanol and make to reach 70% containing alcohol amount, place, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, places, and filters, regulate pH value to 8.0 with sodium hydroxide solution, place, filter, filtrate regulates pH value to 7.0 with 10% hydrochloric acid solution, reclaims ethanol for subsequent use; Herba Epimedii, Rhizoma Polygonati decoct with water secondary, and collecting decoction filters, leave standstill, get supernatant concentration to appropriate, let cool, add ethanol to reaching 65% containing alcohol amount, leave standstill, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leaves standstill, and filters, filtrate regulates pH value to 8.0 with 10% sodium hydroxide, leave standstill, filter, medicinal liquid is for subsequent use; The Radix Astragali decocts with water three times, and collecting decoction filters, and filtrate is concentrated into about 95ml, and medicinal liquid is for subsequent use.Merge above medicinal liquid for subsequent use, stir evenly placement, filter, it is 1.35～1.40(80 DEG C that filtrate is concentrated into relative density) thick paste, add appropriate starch, mix, cold drying, is ground into fine powder, sieves, mix, incapsulate, or above-mentioned thick paste adopts microwave vacuum drying, add appropriate amount of auxiliary materials, mix, incapsulate, altogether make 100 doses, to obtain final product.

The pill preparation of embodiment 4 medicines of the present invention

Radix Polygoni Multiflori Preparata 1300g, Radix Astragali 540g, Rhizoma Polygonati (system) 540g, Herba Epimedii 400g, Fructus Lycii 400g, Radix Salviae Miltiorrhizae 280g;

Above Six-element, Radix Polygoni Multiflori Preparata, Radix Salviae Miltiorrhizae, Fructus Lycii decoct with water three times, collecting decoction, filter, filtrate is concentrated into syrupy shape, lets cool, add ethanol and make to reach 70% containing alcohol amount, place, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, places, and filters, regulate pH value to 8.0 with sodium hydroxide solution, place, filter, filtrate regulates pH value to 7.0 with 10% hydrochloric acid solution, reclaims ethanol for subsequent use; Herba Epimedii, Rhizoma Polygonati decoct with water secondary, and collecting decoction filters, leave standstill, get supernatant concentration to appropriate, let cool, add ethanol to reaching 65% containing alcohol amount, leave standstill, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leaves standstill, and filters, filtrate regulates pH value to 8.0 with 10% sodium hydroxide, leave standstill, filter, medicinal liquid is for subsequent use; The Radix Astragali decocts with water three times, and collecting decoction filters, and filtrate is concentrated into about 95ml, and medicinal liquid is for subsequent use.Merge above medicinal liquid for subsequent use, stir evenly placement, filter, it is 1.35～1.40(80 DEG C that filtrate is concentrated into relative density) thick paste, add appropriate starch, mix, cold drying, is ground into fine powder, sieves, mix, make the watered pill with water pill, or add appropriate refined honey to make honeyed pill, dry, altogether make 100 doses, to obtain final product.

The granule preparation of embodiment 5 medicines of the present invention

Radix Polygoni Multiflori Preparata 700g, Radix Astragali 540g, Rhizoma Polygonati (system) 300g, Herba Epimedii 400g, Fructus Lycii 200g, Radix Salviae Miltiorrhizae 280g;

Above Six-element, Radix Polygoni Multiflori Preparata, Radix Salviae Miltiorrhizae, Fructus Lycii decoct with water three times, collecting decoction, filter, filtrate is concentrated into syrupy shape, lets cool, add ethanol and make to reach 70% containing alcohol amount, place, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, places, and filters, regulate pH value to 8.0 with sodium hydroxide solution, place, filter, filtrate regulates pH value to 7.0 with 10% hydrochloric acid solution, reclaims ethanol for subsequent use; Herba Epimedii, Rhizoma Polygonati decoct with water secondary, and collecting decoction filters, leave standstill, get supernatant concentration to appropriate, let cool, add ethanol to reaching 65% containing alcohol amount, leave standstill, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leaves standstill, and filters, filtrate regulates pH value to 8.0 with 10% sodium hydroxide, leave standstill, filter, medicinal liquid is for subsequent use; The Radix Astragali decocts with water three times, and collecting decoction filters, and filtrate is concentrated into about 95ml, and medicinal liquid is for subsequent use.Merge above medicinal liquid for subsequent use, stir evenly placement, filter, it is 1.35～1.40(80 DEG C that filtrate is concentrated into relative density) thick paste, add appropriate Icing Sugar, dextrin, mix, granulation, or above-mentioned thick paste employing microwave vacuum drying, add appropriate amount of auxiliary materials, mix, granulation, dry, altogether make 100 doses, to obtain final product.

The drop pill preparation of embodiment 6 medicines of the present invention

Radix Polygoni Multiflori Preparata 1000g, Radix Astragali 440g, Rhizoma Polygonati (system) 440g, Herba Epimedii 300g, Fructus Lycii 300g, Radix Salviae Miltiorrhizae 220g;

Above Six-element, Radix Polygoni Multiflori Preparata, Radix Salviae Miltiorrhizae, Fructus Lycii decoct with water three times, collecting decoction, filter, filtrate is concentrated into syrupy shape, lets cool, add ethanol and make to reach 70% containing alcohol amount, place, filter, filtrate adds ethanol again to be made to reach 80% containing alcohol amount, places, and filters, regulate pH value to 8.0 with sodium hydroxide solution, place, filter, filtrate regulates pH value to 7.0 with 10% hydrochloric acid solution, reclaims ethanol for subsequent use; Herba Epimedii, Rhizoma Polygonati decoct with water secondary, and collecting decoction filters, leave standstill, get supernatant concentration to appropriate, let cool, add ethanol to reaching 65% containing alcohol amount, leave standstill, filter, filtrate adds ethanol to be made to reach 80% containing alcohol amount, leaves standstill, and filters, filtrate regulates pH value to 8.0 with 10% sodium hydroxide, leave standstill, filter, medicinal liquid is for subsequent use; The Radix Astragali decocts with water three times, and collecting decoction filters, and filtrate is concentrated into about 95ml, and medicinal liquid is for subsequent use.Merge above medicinal liquid for subsequent use, stir evenly placement, filter, it is 1.20～1.25(80 DEG C that filtrate is concentrated into relative density) thick paste, medicine thick paste+substrate → suspendible or melting → dripping → cooling → wash ball → be dried → select ball → quality inspection → subpackage, makes 100 doses altogether, to obtain final product.

Claims (10)

1. Chinese medicine composition, in an application of preparing in Cure of depression medicine, is characterized in that the crude drug chief component of this Chinese medicine composition is: Radix Polygoni Multiflori Preparata, the Radix Astragali, Rhizoma Polygonati, Herba Epimedii, Fructus Lycii, Radix Salviae Miltiorrhizae.

2. application as claimed in claim 1, is characterized in that, monoamine neurotransmitter in described pharmaceutical composition rising brain.

3. application as claimed in claim 1, is characterized in that, described pharmaceutical composition reduces the monoamine neurotransmitter in blood.

4. application as claimed in claim 2, is characterized in that, the content of 5-hydroxy tryptamine and/or dopamine in described pharmaceutical composition rising Hippocampus.

5. application as claimed in claim 3, is characterized in that, described pharmaceutical composition reduces the content of NE, dopamine, 5-hydroxy tryptamine in blood.

6. application as claimed in claim 1, is characterized in that, described pharmaceutical composition reduces the content of serum thyroliberin, corticosterone and/or β endorphins.

7. application as claimed in claim 1, is characterized in that, described pharmaceutical composition increases neuron number in brain.

8. application as claimed in claim 1, is characterized in that, described pharmaceutical composition alleviates adrenal gland's pathology and changes.

9. application as claimed in claim 1, is characterized in that, described pharmaceutical composition reduces the adrenal cortex thickness of model mouse.

10. application as claimed in claim 1, is characterized in that the crude drug of this Chinese medicine composition consists of:

Radix Polygoni Multiflori Preparata 35-65 weight portion, Radix Astragali 15-27 weight portion, Rhizoma Polygonati 15-27 weight portion, Herba Epimedii 10-20 weight portion, Fructus Lycii 10-20 weight portion, Radix Salviae Miltiorrhizae 7-14 weight portion;

Or, Radix Polygoni Multiflori Preparata 50 weight portions, the Radix Astragali 22 weight portions, Rhizoma Polygonati 22 weight portions, Herba Epimedii 15 weight portions, Fructus Lycii 15 weight portions, Radix Salviae Miltiorrhizae 11 weight portions;

Or, Radix Polygoni Multiflori Preparata extract 35-65 weight portion, Radix Astragali 15-27 extract weight part, Rhizoma Polygonati extract 15-27 weight portion, Herba Epimedii extract 10-20 weight portion, Fructus Lycii extract 10-20 weight portion, Radix Salviae Miltiorrhizae extract 7-14 weight portion.