CN104208383A - Application of fresh lily bulb rehmannia powder in preparation of medicine for treating depressive disorder - Google Patents
Application of fresh lily bulb rehmannia powder in preparation of medicine for treating depressive disorder Download PDFInfo
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- CN104208383A CN104208383A CN201410444555.5A CN201410444555A CN104208383A CN 104208383 A CN104208383 A CN 104208383A CN 201410444555 A CN201410444555 A CN 201410444555A CN 104208383 A CN104208383 A CN 104208383A
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Abstract
The invention relates to an application of fresh lily bulb rehmannia powder in preparation of medicine for treating depressive disorder, which can effectively solve the medication problem that the fresh lily bulb rehmannia powder cannot be used throughout the year to treat depressive disorder. The technical scheme is as follows: the fresh lily bulb rehmannia powder is obtained by mixing fresh lily bulb and rehmannia according to the weight ratio of 1:1, placing the mixture in a refiner to be prepared into serous fluid, drying and pulverizing in a vacuum freeze drier at the temperature of 80 DEG C below zero and vacuum degree of 0.008-0.01mpa to obtain dried powder. The fresh lily bulb rehmannia powder is packaged in a seal manner and stored in a shady and cool place. The fresh lily bulb rehmannia powder is anti-depression, and effectively solves the medication problem that the fresh lily bulb rehmannia powder cannot be used throughout the year to treat depressive disorder. The fresh lily bulb rehmannia powder is rich in materials and easy to prepare. According to the given weight ratio, fresh lily bulb rehmannia powder of any amount can be prepared, is convenient to take, can be applied throughout the year, and is anti-depression and good in effect. The fresh lily bulb rehmannia powder is an innovation of medicine for treating depressive disorder and has huge economic and social benefit.
Description
Technical field
The present invention relates to medicine, the particularly application of a kind of fresh Bulbus Lilii ground bloom in preparation medicament for treatment of depression.
Background technology
Depression is a kind of common frdquently encountered disease, also known as depressive disorder, low for main clinical characteristics with remarkable and lasting mental state, is the main Types of mood disorders.Clinical visible mental state is low unbecoming with its situation, and the downhearted of emotion can from depressed to extremely grieved, and depression of feeling oneself inferior is even pessimistic and worldweary, can have suicidal attempt or behavior; Even occur numb; Some cases have obvious anxiety and mobility intense; The psychotic symptoms such as hallucination, vain hope can be there is in severe patient.Even each outbreak continues at least 2 weeks more than, elder's several years, majority of cases has the tendency of recurrent exerbation, and each outbreak great majority can be alleviated, and part can have residual symptoms or transfer to chronic.Chronic stress can bring out depression.
Chronic stress reaction is called for short chronic stress.Namely refer to that individuality is among a kind of pressure state of long-term chronic.This pressure is not the emergency of some bursts, but allows client feel helpless hopeless a kind of situation, and its final result is also usually " unexpectedly, reasonable again ".
The depression caused by chronic stress is called chronic stress depression, and the medicine for the treatment of chronic stress depression has varied at present, but for various reasons, all unsatisfactory.
Bulbus Lilii, formal name used at school (Lilium brownii var.viridulum Baker) has another name called strong another name for Sichuan Province, Flos Lilii viriduli, Bulbus lilii concoloris, fall celestial, heavily advanced in years, Zhongting, rub sieve, loaded van, in meet flower, Bulbus Lilii Bulbus Allii, cook's Bulbus Allii, Bulbus Allii brain potato, Flos Magnoliae Cocinis etc., it is Liliaceae lilium (formal name used at school: Lilium) perennial herb bulbous plant, sweet, micro-hardship, be slightly cold, enter the heart, lung meridian, nourishing YIN and moistening the lung, clearing away heart-fire for tranquillization, main: deficiency of YIN chronic cough, sputum mixed with blood, the calentura later stage, waste heat is unclear, or the unsuccessful caused fidgets due to deficiency palpitation with fear of feelings will, insomnia and dreamful sleep, absentminded, carbuncle, eczema, the Bulbus Lilii of firm results cleans earth, be called fresh Bulbus Lilii.
Radix Rehmanniae (Classification system: Rehmannia glutinosa (Gaetn.) Libosch.ex Fisch.et Mey.), Scrophulariaceae Radix Rehmanniae belongs to herbaceos perennial, the fresh tuber of scrophulariaceae rehmannia glutinosa plant, clean earth, sweet, bitter, cold, GUIXIN, liver, kidney channel, clearing away heat and promoting production of body fluid, removing heat from blood, hemostasis, for consumption of YIN caused by febrile disease, crimson tongue excessive thirst, send out speckle dermexanthesis, spit blood, epistaxis, laryngopharynx swelling and pain.
So can by mutually composite to fresh Bulbus Lilii and Radix Rehmanniae be the famous lilii and Rehmanniae Decoction of the traditional Chinese medical science, but fresh medicine is by the restriction in time, season, cannot apply throughout the year.The present invention adopts Radix Rehmanniae, fresh Bulbus Lilii by 1:1 proportions, the lyophilization of refiner homogenate final vacuum, then airtight cool place place preserves, and saves the Therapeutic Characteristics of fresh medicine, treatment (chronic stress) patients with depression can be applied to again throughout the year, have no and be publicly reported.
Summary of the invention
For above-mentioned situation, for overcoming the defect of prior art, the object of the present invention is just to provide the application of a kind of fresh Bulbus Lilii ground bloom in preparation treatment (chronic stress) antidepressant agents, and can efficiently solve fresh Bulbus Lilii Dihuang Decoction cannot the medication problem of application for the treatment of depression throughout the year.
The technical scheme that the present invention solves is, fresh Bulbus Lilii, Radix Rehmanniae are mixed with weight ratio 1 ︰ 1, be placed in refiner and make serosity, in vacuum freeze drier dry, powder process under temperature-80 DEG C, vacuum 0.008-0.01mpa condition, obtain dry powder, pack, shady and cool storage, have antidepressant effect, efficiently solving fresh Bulbus Lilii Dihuang Decoction cannot the medication problem of application for the treatment of (chronic stress) depression throughout the year.
Abundant raw material of the present invention, easily prepares, and by the weight ratio provided, can obtain the fresh Bulbus Lilii ground bloom of any amount, taking convenience, can apply throughout the year, have antidepressant effect, effective, be the innovation on treatment (chronic stress) antidepressant agents, economic and social benefit is huge.
Detailed description of the invention
Below in conjunction with embodiment, the specific embodiment of the present invention is elaborated.
The present invention is in concrete enforcement, the application of a kind of fresh Bulbus Lilii ground bloom in preparation medicament for treatment of depression, described fresh Bulbus Lilii ground bloom is, by fresh Bulbus Lilii 600g, the weight ratio of Radix Rehmanniae 600g puts homogenate to refiner, at vacuum freeze drier in temperature-80 DEG C, dry under vacuum 0.008-0.01mpa condition, powder process, obtain dry powder, pack, shady and cool storage, this dry powder has effect of antidepressant, be effective to the medicine preparing treatment (chronic stress) depression, and achieve very satisfied Advantageous Effects through test, related tests data is as follows:
One, animal experiment
1 materials and methods
1.1 experiment material
1.1.1 medicine and reagent
Fresh Bulbus Lilii Radix Rehmanniae dry powder of the present invention; Clomipramine Hydrochloride, friendship nine good fortune pharmaceutcal corporation, Ltd of upper Hisense produces; MDA detection kit, Nanjing is built up Bioengineering Research Institute and is produced; SOD detection kit, Nanjing is built up Bioengineering Research Institute and is produced; O-phthalaldehyde(OPA), Solution on Chemical Reagents in Shanghai purchase and supply 5-linked chemical plant produces; N-butyl alcohol, Tianjin Kai Tong chemical reagent company limited produces; Hydrochloric acid, Laiyang City in pairs Chemical Co., Ltd. is produced; Normal heptane, four-way chemical plant, Tianjin produces; Norepinephrine (NE), Sigma company; Dopamine (DA), Sigma company; 5-hydroxy tryptamine (5-HT), Sigma company; Sodium metaperiodate, good fortune chemical reagent factory in morning in Tianjin is produced; EDTANa2, Bao Xin bio tech ltd produces; Sodium hydrogen phosphate, Tianjin Kai Tong chemical reagent company limited produces; Sodium dihydrogen phosphate, good fortune chemical reagent factory in morning in Tianjin is produced; Sodium acetate, Tianjin chemical reagent three factory produces; Sodium sulfite, Tianjin Kai Tong chemical reagent company limited produces; Dehydrated alcohol, Laiyang City in pairs Chemical Co., Ltd. is produced; Iodine, Tianjin Kermel Chemical Reagent Co., Ltd. produces.
Animals administer dose lonvestion: fresh Bulbus Lilii Radix Rehmanniae (lilii and Rehmanniae Decoction, both ratios are by 1 ︰ 1) quantity every day is by both each 60g/60kg.Rat administration presses 20,15,10 times of quantity.Fresh Bulbus Lilii Radix Rehmanniae flour extraction: be about 6%.
Then rat consumption is: fresh Bulbus Lilii ground bloom: large, medium and small dosage is respectively the fresh powder of 2.4g/fresh powder of kg, 1.8g/fresh powder/kg of kg, 1.2g.
1.2 laboratory animal Wistar rats, 150 ~ 180g, male, provided by Hebei province's medical experiment animal center, credit number: 701022
1.3 experimental apparatus
F-4500 type spectrofluorophotometer, Hitachi, Ltd produces; UV-2000 ultraviolet-uisible spectrophotometer, UNICO(Shanghai) Instruments Co., Ltd. produces; Thermostat water bath, bright instrument plant of Beijing produces; Centrifuge, Beijing Medical Instruments repair shop produces; FA (N)/JA (N) series electronic balance, Shanghai Min Qiao precision instrument company limited produces; Adjustable pipette, Shanghai Lei Bo Analytical Instrument Co., Ltd produces; Calorstat, Beijing forever bright Medical Instruments factory produces; TGL-16G High speed refrigerated centrifuge, Anting Scientific Instrument Factory, Shanghai produces; Rat opens case, self-control.
1.4 statistical procedures
Data analysis SPSS 10.0 for windows statistical software, metric results adopts
represent; Compare employing one factor analysis of variance between measurement data group, ranked data adopt Ridit to analyze.
1.2 experimental technique
1.2.1 experiment grouping and administration
Adopt the scoring of Open-filed method after all rats conform one week, choose the rat 72 that scoring is close, note body weight simultaneously, sucrose solution consumption is without marked difference, animal is divided into 6 groups at random, wherein makes rat chronic Stress Depression Model for 5 groups, another 1 group is blank group.5 groups, modeling type gavages Clomipramine Hydrochloride suspension (positive control respectively, dosage is 20mg/kg, 2mg/m1 is made into before use with normal saline, 1ml/100g), large, medium and small dosage fresh Bulbus Lilii Radix Rehmanniae dry powder suspension (2.4g/kg, 1.8g/kg, 1.2g/kg, 0.24g/m1,0.18g/m1,0.12g/m1 is respectively containing crude drug concentration, 1ml/100g, be equivalent to 20,15,10 times of quantity respectively), model group and blank group gavage the normal saline of same volume, and administration volume is 1ml/100g.The administration simultaneously of modeling type, administration every day 1 time, successive administration 21d.
1.2.2 modeling method
5 raisings of the every cage of normal group, normal diet is drunk water, and does not give outside any stimulation.5 groups, modeling type is every cage and raises 1 rat, and every day gives different stimulated at random.7 kinds of stress factors are applied in 21d by random method: 1. illumination (24h) all night; 2. fasting (24h); 3. water (24h) is prohibited; 4. 4 DEG C of cold-wate swimming 5min, 5. 1min presss from both sides tail, 6. 5min high speed horizontal oscillations; 7. 2h behavior restriction.Give a kind of stimulation every day at random, often kind of stimulation can not occur continuously, continuous 21d.
1.2.3 chronic stress depression model schedule is made
1.2.4 laboratory animal process
1.2.4.1 sucrose solution intake is tested
After model preparation starts first 1 day and terminates, the 1st day morning tested each group of rat 1% sucrose water consumption, prohibits water 12h, measure sucrose water intake in 1h before test.
1.2.4.2 Open-filed method is tested
Experimental provision is that cube opens case, high 40cm, and length and width are respectively 80cm, and perisporium, bottom surface are black, and bottom surface is made up of 25 pieces of area equation, divides with white line.During experiment, rat is placed in the grid of Chang Xiang center, observe horizontal anomalous movement (crossing) score that rat passes through bottom surface block number (grid that four paws all enters can count) in 3min, and Vertical movements (rearing) score of rear number of times (two fore paws soar or seek connections with wall).Feces thoroughly must be removed after each experiment is complete.Every animal starts to terminate the latter 1 day morning with experiment in first 1 day in experiment and measures 1 time, respectively organizes score difference.This experiment is carried out in quiet room.
1.2.4.3 the mensuration of MDA level and erythrocyte sod vigor in blood plasma
After Behavior test, rat eye socket gets blood, heparin sodium anticoagulant, the centrifugal 10min of 3000r/min, separated plasma, measures MDA level and erythrocyte sod vigor in blood plasma respectively by test kit description.
1.2.4.3.1 MDA level determination in blood plasma
MDA test kit test philosophy: the malonaldehyde (MDA) in lipid peroxide catabolite can with thiobarbituricacidα-(TBA) condensation, formed red product, go out to have maximum absorption band at 532nm.Because of substrate for thiobarbituricacidα-(Thibabituric Acid TBA) claims TBA method in this way.
100T test kit composition and preparation:
Reagent 1: liquid 20ml × 1 bottle, room temperature preservation.(can solidify when it is cold, before each test, suitably water-bath is heated with accelerate dissolution, can apply until transparent.)
Reagent 2: liquid 6ml × 1 bottle, the used time every bottle adds the mixing of 340ml distilled water, 4 DEG C of cold preservations.
Reagent 3: powder × 1, powder joins in the hot distilled water 60ml of 90 DEG C ~ 100 DEG C by the used time, (can suitably heat in course of dissolution), complements to 60ml after fully dissolving with distilled water, mixing, the reagent lucifuge cold preservation prepared.
Reagent 4: standard substance: 10nmol/m1 tetraethoxypropane 5ml × 1 bottle, 4 DEG C of cold preservations.
Operational approach: the preparation of mix reagent: total sample number n is relaxed 4 amounts and be multiplied by 1 respectively, 2, No. 3 reagent, by amount of calculation, 3 kinds of reagent are mixed.Mix reagent total amount=[(n+4 note) × No. 1 reagent 0.15ml]+[(n+4) × No. 2 reagent 3ml]+[(n+4) × No. 3 reagent 1ml].Note: the number adding standard pipe and standard blank tube by the sample number of required detection, then relax 2 (avoiding being drawn onto last amount of reagent inadequate).Mix reagent needs matching while using, in table 1.
The preparation of table 1 mix reagent
Swirl mixing device mixes, test tube mouth preservative film is tightened, and stings an aperture with syringe needle, 95 DEG C of water-baths (or uncap with pot boil) 40min, flowing water cooling after taking out, then 3500 ~ 4000r/min, centrifugal 10min, gets supernatant, 522nm place, 1cm optical path, distilled water returns to zero, and surveys each pipe absorbance.
1.2.4.3.2 the mensuration of superoxide dismutase (SOD) vigor in erythrocyte
Measuring principle: produce ultra-oxygen anion free radical (O2.-) by xanthine oxidase response system, the latter is oxidized azanol and forms nitrite, presents aubergine, survey its absorbance with visible spectrophotometer under the effect of developer.When containing SOD in sample, then there is narrow spectrum inhibitory action to ultra-oxygen anion free radical, the nitrite of formation is reduced, and the absorbance measuring pipe during colorimetric, lower than the absorbance of control tube, can obtain the SOD vigor in sample by formulae discovery.
The composition of reagent and preparation:
Reagent 1: stock solution: 10ml × 1 bottle (when it is cold or put refrigerator and have partially crystallizable and separate out, use again after needing hot bath to dissolve); The preparation of liquid applied by reagent 1: used time every bottle adding distil water is diluted to 100ml, 4 DEG C of preservations.
Reagent 2: liquid 10ml × 1 bottle, 4 DEG C ~ 10 DEG C preservations.
Reagent 3: liquid 10ml × 1 bottle, 4 DEG C ~ 10 DEG C preservations.
Reagent 4: liquid 350 μ l × 2,4 DEG C of preservations, can not be freezing; No. 4 diluent 10ml × 1 bottle, 4 DEG C of preservations.Used time both presses 1:14 dilution, can once dilute, also can Fractional dilution.The reagent prepared 4 DEG C preservation, can not be freezing.2 ~ 3 months can be preserved after preparing.Note: suction nozzle used is preferably special and disinfected.
Reagent 5: powder × 1, the used time adds 70 DEG C ~ 80 DEG C hot distilled water 75ml and dissolves rear for subsequent use, if moisture evaporation reduces in heating process, now must be supplemented to 75ml with distilled water, the 4 DEG C of cold preservations of the reagent lucifuge after preparing.
Reagent 6: powder × 1, used time adding distil water 75ml is for subsequent use after dissolving, the 4 DEG C of stored refrigerated of the reagent lucifuge after preparing.
The preparation of developer: according to reagent 5: reagent 6: the volume ratio of glacial acetic acid=3:3:2 is made into developer, 4 DEG C of lucifuge cold preservations.Fully mix with swirl mixing device, make 37 DEG C of water bath with thermostatic control 40min and mix, room temperature places 10min, and in wavelength 550nm place, 1cm optical path cuvette, distilled water returns to zero, and colorimetric, in table 2.
Total SOD (T-SOD) the vitality test step of table 2
1.2.4.4 on the impact of monoamine neurotransmitter in brain
Put to death after rat extracting blood, get brain, preparation brain tissue homogenate, with the content of monoamine neurotransmitter 5-HT, NE, DA in fluorescence spectrometry brain tissue homogenate.
1.2.4.4.1 pre-treatment
Weigh after cerebral tissue being removed blood film, olfactory bulb and cerebellum, in ice-water bath, make homogenate with 5 times amount acidify n-butyl alcohol (to add concentrated hydrochloric acid 0.85m1 in 1000m1 n-butyl alcohol).Vortex vibration 5min, centrifugal 5min (3000r/min).Get supernatant 2.5m1, put into another band plug centrifuge tube, add normal heptane 5m1 and 0.1mol/L hydrochloric acid 1.2m1, vortex vibration 5min, centrifugal 5min, obtain layering solution (aqueous phase I contains 5-HT, DA, NE).
1.2.4.4.2 typical shelf liquid
5-HT, DA, NE are mixed with 500 μ g/m1 as normal storage liquid with 0.0lmol/L hydrochloric acid respectively.Deposit in 2 DEG C of refrigerators.
1.2.4.4.3 Standard Applying Solution
Face the used time and respectively get above-mentioned stock solution 0.25m1, be diluted to 100m1 Standard Applying Solution with 0.0lmol/L HCL.Each standard concentration 1.25 μ g/m1.
1.2.4.4.4 the mensuration of 5-HT, NE and DA in cerebral tissue
1.2.4.4.4.1 the mensuration of 5-HT
Water intaking phase I0.5m1 adds 0.5% cysteine (fresh configuration) 0.1m1,0.005% o-phthalaldehyde(OPA) 3m1 (being temporarily made into l0mol/L HCL), 0.02% sodium metaperiodate 0.lml, and boiling water bath 10min, is cooled to room temperature, measures.0.5% cysteine, prepares with 0.1mo1/L hydrochloric acid.
Standard pipe and blank tube
Standard Applying Solution 0.5ml got by standard pipe, and blank tube gets double distilled water 0.5m1; HCl dropwise n-butyl alcohol 4.5m1, vortex vibration 5min, centrifugal 5min (3000r/min).Get supernatant 2.5m1, put into another band plug centrifuge tube, add normal heptane 5m1 and 0.1mol/L hydrochloric acid 1.0ml, vortex vibration 5min, centrifugal 5min, obtain layering solution (aqueous phase I is containing 5-HT, DA, NE).
Water intaking phase I 0.5m1 adds 0.5% cysteine 0.lml, 0.005% o-phthalaldehyde(OPA) 3m1 (being made into l0mol/L HCL), 0.02% sodium metaperiodate 0.lml, and boiling water bath 10min, is cooled to room temperature, measures.
5-HT maximum emission wavelength (em) 475nm, maximum excitation wavelength (ex) 355nm.
1.2.4.4.4.2 the mensuration of NE and DA
Water intaking phase I 0.5m1, adds 1/15mol/L phosphate buffered saline(PBS) (PH7.2) 1.7m1,0.1mol/L EDTANa
2(with the preparation of 1mo1/L sodium acetate solution, adjusting PH to 7.0 with l0mol/L NaOH) 0.4m1, adds iodine reagent 0.lml, leave standstill 2min, add alkaline sodium sulfite solution 0.5m1 (25% sodium sulfite 1.0m1,5mol/L sodium hydroxide 9m1 mixes temporarily), leave standstill 2min.Add 6mo1/L HAc liquid 0.5m1, boiling water bath 20min, is cooled to room temperature, measures NE respectively, DA.
Iodine reagent: preparation 0.02mo1/L iodine liquid and 5% sodium iodide (with 70% dissolve with ethanol), before use with 1 ︰ 1 volume ratio mixing.
Standard pipe and blank tube: Standard Applying Solution 0.5ml got by standard pipe, blank tube gets double distilled water 0.5ml, with acidify n-butyl alcohol 4.5m1, vortex vibration 5min, centrifugal 5min (3000r/min).Get supernatant 2.5ml, put into another band plug centrifuge tube, add normal heptane 5m1 and 0.1mo1/L hydrochloric acid 1.2m1, vortex vibration 5min, centrifugal 5min, obtain layering solution (aqueous phase I contains 5-HT, DA, NE).
Water intaking phase I 0.5m1, add 1/15mol/L phosphate buffered saline(PBS) (PH7.2) 1.7m1,0.1mo1/L EDTANa2 (prepares with lmol/L sodium acetate solution, PH to 7.0 is adjusted with 10mol/L NaOH) 0.4m1, add iodine reagent 0.2m1, leave standstill 2min, add alkaline sodium sulfite solution 0.5m1 (25% sodium sulfite 1.0m1,5mol/L sodium hydroxide 9m1 mixes temporarily), leave standstill 2min.Add 6mol/LHAc liquid 0.5m1, boiling water bath 20min, is cooled to room temperature, measures NE respectively, DA.
NE maximum emission wavelength (em) 475nm, maximum excitation wavelength (ex) 385nm; DA maximum emission wavelength (em) 370nm, maximum excitation wavelength (ex) 322nm.
1.2.4.4.4.3 computing formula
Sample monoamine transmitter contents (μ g/g cerebral tissue)=(Ri-Rib)/(Rr-Rrb) × 0.5 (ml) × 1.25 (ug/m1) ÷ real EEG weight
Ri: be test sample fluorescence reading; Rib: be test sample blank fluorescence reading;
Rr: be reference substance fluorescence reading; Rrb: be the blank reading of reference substance.
1.2.4.5 on the impact of Thymus and spleen tissue morphology
Put to death after rat extracting blood, dissect, get Thymus and spleen, 10% formalin is fixed, and through embedding, section, dyeing, microscopy, observes the impact on immune organ.
2. experimental result
2.1 impacts on rat chronic Stress Depression Model body weight
In experiment beginning, experiment the 1st week, test the 2nd week and test the 3rd week, give each group of rat weight respectively, respectively the change of group rat body weight in modeling type and administration process, the results are shown in Table 3.
Table 3 fresh Bulbus Lilii ground bloom is on the impact of the rat chronic Stress Depression Model scale of construction
* with model group than P < 0.05, * * with model group than P < 0.01
Can find out from upper table, respectively organize no significant difference between rat body weight before administration, illustrate that grouping evenly; With blank group ratio, model group rats lost weight from the 1st week, is starkly lower than blank group (P < 0.05), illustrates that rat chronic Stress Depression Model weight ratio normally reduces to the 3rd week body weight.With model group ratio, all increase is had the 1st week and the 2nd week each administration group body weight, at the 3rd week, heavy dose of group fresh Bulbus Lilii ground bloom group and small dose group fresh Bulbus Lilii ground bloom group all can make rat body weight obviously increase (P < 0.05), the trend that other each administration groups also can make rat body weight increase.
2.2 impacts on rat chronic Stress Depression Model sucrose solution consumption
Terminate in experiment, measure each group of rat respectively in experiment end intake of sucrose water in 1h after 1 day, the results are shown in Table 4.
Table 4 fresh Bulbus Lilii ground bloom is on the impact of rat chronic Stress Depression Model sucrose solution consumption
* with model group than P < 0.05, * * with model group than P < 0.01
Can find out from upper table, with blank group ratio, model group rats sucrose solution consumption significantly reduces (P < 0.01), illustrates and makes the success of rat chronic Stress Depression Model.With model group ratio, large, medium and small dosage fresh Bulbus Lilii ground bloom group and Clomipramine Hydrochloride group all significantly can increase rat sucrose solution consumption (P < 0.01).
2.3 on the impact of rat chronic Stress Depression Model horizontal movement score with the score that moves both vertically
By the experimental technique of Open-filed experiment regulation, observe each group of rat horizontal movement number of times and to move both vertically score in 3min in 1 day after experiment terminates, the results are shown in Table 5.
Table 5 fresh Bulbus Lilii ground bloom is on the ethological impact of rat chronic Stress Depression Model
* with model group than P < 0.05, * * with model group than P < 0.01
Can find out from upper table, with blank group ratio, in model group rats 3min, horizontal movement score and the score that moves both vertically all significantly reduce (P < 0.01), illustrate and make the success of rat chronic Stress Depression Model.With model group ratio, in, low dose of fresh Bulbus Lilii ground bloom group and Clomipramine Hydrochloride group all obviously can increase horizontal movement score in rat 3min (P < 0.05), heavy dose of group fresh Bulbus Lilii ground bloom group has the trend increasing horizontal movement score in rat 3min; Dosage fresh Bulbus Lilii ground big or middle bloom group obviously can increase in rat 3min the score that moves both vertically (P < 0.05), and low dose of fresh Bulbus Lilii ground bloom group and Clomipramine Hydrochloride group all significantly can increase in rat 3min the score that moves both vertically (P < 0.01).
2.4 impacts on MDA level in rat chronic Stress Depression Model erythrocyte sod vigor and blood plasma
By the method that SOD test kit and MDA test kit provide, detect MDA level in chronic stress depression model Rat Erythrocytes SOD vigor and blood plasma, the results are shown in Table 6.
Table 6 fresh Bulbus Lilii ground bloom is on the impact of MDA level in rat chronic Stress Depression Model erythrocyte sod vigor and blood plasma
* with model group than P < 0.05, * * with model group than P < 0.01
Can find out from upper table, with blank group ratio, model group rats erythrocyte sod vigor obviously reduces (P < 0.05), and blood plasma MDA level has increase trend, illustrates that rat chronic Stress Depression Model oxidation resistance obviously reduces.With model group ratio, dosage fresh Bulbus Lilii ground big or middle bloom group obviously can increase rat model erythrocyte sod vigor (P < 0.05), Clomipramine Hydrochloride group significantly can increase rat model erythrocyte sod vigor (P < 0.01), and low dose of fresh Bulbus Lilii ground bloom group has the trend increasing rat model erythrocyte sod vigor; Large, medium and small dosage fresh Bulbus Lilii ground bloom group has the trend reducing rat model blood plasma MDA level.
2.5 impacts on monoamine neurotransmitter level in rat chronic Stress Depression Model brain homogenate
Application spectrofluorophotometer method, by 5-HT, NE and DA test kit description, measures 5-HT, NE and DA level in brain tissue homogenate respectively, the results are shown in Table 7.
Table 7 fresh Bulbus Lilii ground bloom is on the impact of monoamine neurotransmitter level in rat chronic Stress Depression Model brain
* with model group than P < 0.05, * * with model group than P < 0.01
Can find out from upper table, with blank group ratio, in model group rats cerebral tissue, NE level, 5-HT level and DA level all significantly reduce (P < 0.01), illustrate and make the success of rat chronic Stress Depression Model.With model group ratio, large, medium and small dosage fresh Bulbus Lilii ground bloom group and Clomipramine Hydrochloride group all significantly can increase NE level in rat cerebral tissue, 5-HT level and DA level (P < 0.01).
2.6 impacts on rat chronic Stress Depression Model thymic tissue form
Get rat chest gland and spleen, through fixing, embedding, section, dyeing, microscopy, observe fresh Bulbus Lilii ground bloom to the impact of rat chronic Stress Depression Model immune organ and tissue form.
Observe thymus under an optical microscope visible, blank group rat chest gland lobule leaflet is clear, and cortex and medullary substance are demarcated clear, and cortilymph cell is intensive; Model group rats atrophy of thymus gland, the boundary of cortex medullary substance is not too clear, and cortex is obviously thinning, and lymphocyte quantity and density all reduce; Clomipramine Hydrochloride group rat chest gland lobule leaflet is clear, and cortex comparatively model thickens to some extent, and cell density obviously increases; Bloom group rat chest gland leaflet is clear on heavy dose of fresh Bulbus Lilii ground, and cortical thickness comparatively model group thickens, and cell density and quantity increase to some extent; Middle dosage fresh Bulbus Lilii ground bloom group rat chest gland is clear, and cortex comparatively model group obviously thickens, and cell density and quantity obviously increase; Low dose of fresh Bulbus Lilii ground bloom group Mouse Thymic Cortex obviously thickens, and cell density and quantity obviously increase.The results are shown in Table 8.
Table 8 fresh Bulbus Lilii ground bloom is on the impact of rat chronic Stress Depression Model thymus pathological change
*: represent and compare P < 0.05 with model group; *: represent and compare P < 0.01 with model group
Can find out from upper table, with blank group ratio, model group rats animal thymus cortical thickness and cortex cell number all significantly reduce (P < 0.01), illustrate and make atrophy of thymus gland after rat chronic Stress Depression Model.With model group ratio, large, medium and small dosage fresh Bulbus Lilii ground bloom group and Clomipramine Hydrochloride group all significantly can increase Mouse Thymic Cortex thickness (P < 0.01), significantly increase Mouse Thymic Cortex lymphocyte number (P < 0.01).With in, the ground knot of bloom group to the atrophy of thymus gland anti-effect of low dose of fresh Bulbus Lilii as well.
Observe spleen under an optical microscope visible, the boundary of blank group Rats Spleen red white pulp clearly, snius lienis and spleen little all normal; The boundary of model group rats spleen red white pulp is not too clear, and splenic nodule obviously reduces, and lymphocyte obviously reduces; The boundary of Clomipramine Hydrochloride group Rats Spleen red white pulp is clear, and splenic nodule and lymphocyte number comparatively model group increase to some extent and increase; Heavy dose of fresh Bulbus Lilii ground bloom group Rats Spleen splenic nodule comparatively model increases, and lymphocyte density increases, lymphocytosis; Middle dosage fresh Bulbus Lilii ground bloom group Rats Spleen splenic nodule comparatively model group obviously increases, and cell density increases, lymphocytosis; Low dose of fresh Bulbus Lilii ground bloom group Rats Spleen splenic nodule obviously increases, and cell density increases, lymphocyte showed increased.The results are shown in Table 9.
Table 9 fresh Bulbus Lilii ground bloom is on the impact of rat chronic Stress Depression Model spleen pathological change
*: represent and compare P < 0.05 with model group; *: represent and compare P < 0.01 with model group
Can find out from upper table, with blank group ratio, model group rats spleen trifle significantly reduces (P < 0.01), and lymphocyte number significantly reduces (P < 0.01), illustrates and makes spleen atrophy after rat chronic Stress Depression Model.With model group ratio, large, medium and small dosage fresh Bulbus Lilii ground bloom group and Clomipramine Hydrochloride group all can significantly make Rats Spleen spleen trifle increase (P < 0.01), spleen cortilymph cell number is all significantly increased (P < 0.01).With low dose of fresh Bulbus Lilii ground bloom group to the antagonism of spleen atrophy as well.(P＜0.01)。
3 conclusions
Shown by above-mentioned test, integrated use stress support two kinds of classical ways with orphan, utilize long-term Unpredictability stress, cause and lonely support animal depressive state.And with drug test of the present invention, monoamine transmitters 5-HT, NE, DA content in chronic stress rat depression model plasma SOD vigor, brain is had a significant effect.Compare with model group, SOD vigor can be improved, significantly increase brain mediator content.Experiment prompting, the antidepressant effect of fresh Bulbus Lilii ground bloom may be relevant with monoamine neurotransmitter (5-HT, NE, DA) content in raising body scavenging free radicals, oxidation resistance and increase brain.
The each dosage group of fresh Bulbus Lilii ground bloom has improvement result in various degree to the damage that thymus, spleen in chronic stress rat modeling process occur.Prompting fresh Bulbus Lilii ground bloom antidepressant effect may realize by improving Organism of Rats immunity.
Experiment proves, compared with blank, model group rats level passes through lattice number, number of times of standing, sucrose solution consumption all significantly reduce, and when the 3rd week, rat body weight obviously reduces.These performances illustrate that Animal performance go out depressive state, hebetude and anhedonia, changes, the forfeiture of interest or pleasant sensation has similarity to a certain degree, points out copying of rat depression model to be successful with the psychomotor in depression clinical diagnosis.Result shows, compared with model group, the level that fresh Bulbus Lilii ground bloom respectively organizes rat passes through lattice number, number of times of standing, sucrose solution consumption, rat body weight all have increase, and fresh Bulbus Lilii is described bloom can improve the depressive symptom of chronic stress rat, has antidepressant effect.And the Advantageous Effects do not expected is achieved through clinical practice.
Two, clinical trial
Patient through suffering from slightly (chronic stress) depression to 158 examples treats with fresh Bulbus Lilii ground of the present invention bloom, every day, quantity pressed 120g/60kg, namely the every 60kg of human body is heavy, every day, consumption was 120g, can sooner or later service once, each each 20g, curative effect is evaluated, statistical confirmation, in 158 routine patients after 30 days, only have its depression of 1 people do not have be improved significantly or improve, all the other have symptom in various degree to improve, and there are raising in various degree and improvement quality of life, and effective percentage is up to more than 99%, its effect good, did not formerly expect.
In a word, can clearly be found out by above-mentioned test, fresh Bulbus Lilii Radix Rehmanniae powder, preparation method thereof of the present invention is simple, abundant raw material, open up the medical value of fresh Bulbus Lilii, Radix Rehmanniae compositions, achieve the application of fresh Bulbus Lilii ground bloom in preparation treatment chronic stress antidepressant agents, be the innovation on treatment chronic stress antidepressant agents, economic and social benefit is huge.
Claims (1)
1. the application of a fresh Bulbus Lilii ground bloom in preparation medicament for treatment of depression, this fresh Bulbus Lilii ground bloom is mixed with weight ratio 1 ︰ 1 fresh Bulbus Lilii, Radix Rehmanniae, be placed in refiner and make serosity,, powder process dry in vacuum freeze drier under temperature-80 DEG C, vacuum 0.008-0.01mpa condition, obtain dry powder, realize the application of fresh Bulbus Lilii Dihuang Decoction throughout the year in medicament for treatment of depression.
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Non-Patent Citations (2)
| Title |
|---|
| 苗明三等: "中药鲜药加工方法探讨", 《时珍国医国药》 * |
| 苗晋鑫等: "鲜怀地黄粉对小鼠抑郁模型的影响", 《中医学报》 * |
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Application publication date: 20141217 |