CN103108639B - Composition of active ingredient of traditional chinese medicine and use thereof - Google Patents

Abstract

Disclosed is a composition of an active ingredient of traditional Chinese medicine, comprising geniposide or jasminoidin, berberine, total saponins of panax ginseng or a composition of ginsenoside monomer. Also disclosed is the use of the pharmaceutical composition in preparing drugs for treating or preventing brain and nerve damage, neurodegenerative encephalopathy, bacterial or viral encephalitis, or depression.

Description

A kind of active ingredient of Chinese herbs composition and use thereof

Technical field

The present invention relates to a kind of active ingredient of Chinese herbs compositions, it comprises geniposide or jasminoidin, berberine, Radix Ginseng total saponins or ginsenoside monomer or ginsenoside monomer compositions.The invention still further relates to the purposes of said composition in the medicine of preparation treatment or prevention brain and nerve injury, nervus retrogression encephalopathy, bacillary or viral encephalitis or depression.

Background technology

Theory of Chinese medical science great attention to whole idea and determination for the treatment of based on pathogenesis obtained through differentiation of symptoms and signs, Traditional Chinese medical theory is thought, blood stasis due to qi deficiency is apoplexy the important pathogenesis, and the deficiency of vital energy is primary disease basis, and blood stasis is primary disease basic pathogenesis, is the core that development occurs primary disease, and blood circulation promoting and blood stasis dispelling is the basic rule for the treatment of of treatment primary disease.Blood stasis due to qi deficiency disease only sees the apoplexy sequela phase in traditional understanding, Blood rheology is also only for sequela patient, Qing Dynasty well-known doctor Wang Qingren BUYANG HUANWU TANG represents prescription (Zhang Meikui, Yin Ling, the progress of Zhang Xuewen Buyang Huanwu Decoction in Treating cerebral infarction, China's TCM basis medical journal, 2002,8 (9): 74-78).And there is not been reported to adopt QI invigorating, removing toxic substances method for the treatment of and relevant treatment by Chinese herbs brain and nerve injury disease.

Alzheimer disease (senile dementia, SD), also known as Alzheimer (Alzheimer disease, AD) be a kind of central nervous system degenerative disease damaged based on Progressive symmetric erythrokeratodermia cognitive disorder and memory ability, and with many-sided exceptions such as motion in various degree, cognition, language and personalities.The clinical manifestation of AD is dementia syndrome, with hypomnesis, personality degeneration for feature.The main pathological change of AD is cerebral cortical atrophy, neurocyte and synapse is lost in a large number, neurofibrillary tangle, extracellular senile plaque and cerebrovascular amyloidosis.At present basic Therapeutic Method be there is no to senile dementia.Current treatment alzheimer disease molecular mechanism and relevant plant amedica have: 1) acetylcholinesteraseinhibitors inhibitors Huperzine A; 2) excitatory amino acid and peptide class mediator regulator, as leaf autumn alkali; 3) amyloid-beta (A β) processbearing astrocyte and deposition is disturbed, as ginkgetin, salvianolic acid, emodin, Sarsasapogenin etc.4) antioxidation and scavenging free radicals medicine, the extract tea polyphenols, Folium Ginkgo extract, ginsenoside, bilobalide etc. of radix Salviae Miltiorrhizae water extract salvianolic acid, Folium Camelliae sinensis; 5) microcirculation is improved, as TANSHINONES and danshensu; 6) calcium antagonist, as apigenin.

Vascular dementia (Vascular Demenia, be called for short VD) be because a series of cerebrovascular factor causes the general name of the chronic brain syndrome caused by brain tissue damage, be occur on cerebrovascular basis based on memory, cognitive dysfunction, or a kind of disease of acquired intelligent persistence infringement with language, space technical ability and emotion or personality disorder.Clinical in hypomnesis, the Progressive symmetric erythrokeratodermia obstacle of abilities such as calculating, judge, be directed and spirit, behavior is feature, is the commonly encountered diseases of mid-aged population, frequently-occurring disease.The definite pathomechanism of VD still imperfectly understands at present, it is generally acknowledged, cerebrovascular disease causes local brain tissue inflammation, metabolism disorder and oxidative stress loss, damage the cerebral tissue of enough capacity, the senior function of nervous system, particularly focus such as grievous injury memory and cognition are more prone to occur dementia symptom when relating to frontal lobe, temporal lobe and limbic system.Treatment VD Chinese medicine compound have: Rhizoma Coptidis toxic materials clearing away decoction, DANGGUI SHAOYAO SAN, dihuang drink, TIANZHI KELI, clever holy capsule, ginseng fiber crops Cognex, LIUWEI DIHUANG TANG, add taste GUIPI TANG, BUYANG HUANWU TANG etc.And there is not been reported to adopt QI invigorating, removing toxic substances method for the treatment of and relevant Chinese medicine.

Parkinson disease (Parkinson disease, PD) have another name called Parkinsonism, and in being, the common central system disorder of old people, clinical manifestation is static tremor, bradykinesia, myotonia and posture abnormal gait etc.Pathogenesis is dopamine (DA) nervous system pathological changes in maincenter nigro-striatal pathway, causes striatum DA to lack, causes somatic movement dysfunction.DOPAMINE CONTENT IN RABBIT reduces, and acetylcholine will superexcitation, two kinds of mediator disequilibriums, causes neurally can't help producing tetanic, the symptom such as to tremble with muscular tissue.At present, parkinson disease there is no radical cure medicine, and Western medicine levodopa (madopar), Ergolactin, Ergota card woods etc. are clinical common medicines, but effect is unstable, and toxic and side effects is comparatively large, and late result is not also affirmed.The clinical common method for the treatment of of Chinese medicine and common drug as follows: 1) tonifying liver is empty, town liver endogenous wind stopping: adopt Rhizoma Gastrodiae, Colla cornus cervi, Radix Asparagi, Radix Ophiopogonis, Carapax Trionycis, Carapax et Plastrum Testudinis, Radix Paeoniae Alba, Radix Glycyrrhizae etc. to be used as medicine; 2) the kidney invigorating is empty, replenishing essence QI invigorating: Fructus Corni, Fructus Schisandrae Chinensis; 3) replenishing vital essence and blood, tonifying brain with essence: adopt Herba Cistanches, Radix Angelicae Sinensis, Arillus Longan, Fructus Jujubae etc.; 4) blood circulation promoting and blood stasis dispelling: Radix Notoginseng, Squama Manis, Radix Salviae Miltiorrhizae, Scorpio.Chen Jianzong etc. find that the Chinese medicine of the nourishing liver and kidney of the compositions such as Fructus Lycii, Herba Cistanches, Radix Polygoni Multiflori can improve PD rat brain striatum DA; 3,4-dihydroxyphenyl acetic acid (DOPAC); NE; 5-HT equal size; and (Chen Jianzong, Guo Jianying, grandson wait quietly, and Vascular Parkinsonism 40 example is treated in the ruling by law of nourishing liver and kidney calming wind and dredging collateral, Journal of Traditional Chinese Medicine to have certain protective effect to nerve nucleus such as black substance, locus coeruleus, nucleus raphes dorsalises; 2002,43 (7): 528-529).The researchs such as what builds up find by Fructus Lycii, Herba Taxilli, Rhizoma Gastrodiae, Ramulus Uncariae Cum Uncis, what Bombyx Batryticatus etc. formed has enriching yin and nourishing kidney, the Chinese medicinal formulae of Tongluo Jiedu effect obviously can reduce PD rat brain release active oxygen (ROS), malonaldehyde (MDA) content, make glutathion (GSH), glutathione peroxidase (GSH-PX), the Free-radical ring opening polymerization content such as superoxide dismutase (SOD) rise, there is raising antioxidant ability of organism, the effect alleviating radical pair body injury (can be built up, Yuan Canxing, Li Yaming etc., nourishing the liver and kidney Tongluo Jiedu Chinese medicine is on the impact of Rat Model with Parkinson Disease response to oxidative stress, Chinese Journal of New Drugs and Clinical Remedies, 2003, 22 (3): 160-163).Xu Li etc. observe endogenous wind stopping relieving convulsion capsule (Radix Angelicae Sinensis, the Radix Paeoniae Alba, Scolopendra, the nourishing YIN and benefiting blood such as Scorpio, endogenous wind stopping dredging collateral Chinese medicine forms) treatment qi and blood deficiency experimental PD mice, find that it can significantly increase DA in PD mouse brain, DOPAC, the content of 4-hydroxy-3-methoxy-.alpha.-toluic acid. (HVA), and mouse limb can be reduced to tremble intensity, shorten and tremble the persistent period, therefore think that endogenous wind stopping relieving convulsion capsule has preventive and therapeutic effect (Xu Li to PD, Wei Cuie, Liu Jianxun etc., Xifeng Zhijing Capsule is to the Parkinsonian preventive and therapeutic effect of mouse experiment, new Chinese medicine and clinical pharmacology, 2005, 2:87-90).And there is not been reported to adopt QI invigorating, removing toxic substances method for the treatment of and relevant Chinese medicine.

Depression is commonly referred to as a kind of dysthymic disorder, mainly, bradyphrenia, language and behavior low with mental state are reduced to the syndrome of principal character, and main manifestations is depressed, interest attenuating, pessimism, retardation of thinking, shortage initiative, self-accusation are poor from crime, diet, sleep, worry oneself to suffer from various disease, feel whole body many places discomfort, suicidal thought and behavior can appear in severe patient.Depression has the features such as sickness rate is high, prevalence is high, relapse rate is high, homicide rate is high and awareness is low, treatment rate is low.The depression cause of disease is complicated, and the pathomechanism hypothesis of existing research at present has several as follows: 1) Monoamine Metabolism is abnormal, and depression is due in brain caused by some position monoamine neurotransmitter deficiency; 2) function of receptors changes, and antidepressants suppress presynaptic monoamine transmitters picked-up to be acute effect, and it is then a kind of chronic adaptations that postsynaptic receptor sensitivity reduces; 3) neuroendocrine disturbance, research finds that the dysfunction of depressed generation and hypothalamus one hypophysis one hypothalamic pituitary adrenal axis (HPA), hypothalamic-pituitary-thyroid axis (HPT), Hypothalamus-pituitary-growth hormone axle (HPGH) etc. and serum cholesterol level reduce, and serum estrogen levels declines relevant.Current antidepressants are mainly containing 3 kinds of characteristics in varying degrees, namely have excitation to psychomotor, strengthen drive, enliven emotion and to angst resistance effect.It is mainly divided into five large classifications: oxidase inhibitor, tricyclic antidepressant (TCAs, as Tatinol), tetracyclic antidepressants, novel antidepressant is if selectivity 5-is through reuptake inhibitors (SSRIs, as fluoxetine), selectivity NRI, 5-hydroxy tryptamine and norepinephrine reuptake double inhibitor (SNRIs, as venlafaxine), norepinephrine and specific 5-are through reuptake inhibitors (NaSSAs, as mirtazapine), norepinephrine and dopamine reuptake inhibitor, plant amedica extract etc.But mostly have that narrow, the side effect of antidepressant spectrum is large, medicine valency is high and the feature such as easy recurrence.

Meningitis can be divided into bacillary and aseptic two class.Bacterial meningitis is the inflammation caused due to bacteriological infection pia mater encephali, and be the serious infectious disease of a kind of intracranial, often can occur together encephalitis and brain abscess.Say from nosetiology angle and comprise two large classes, a class is the meningitis (being called for short: change brain) that various suppurative bacterium infects, as: the meningitis that meningococcus, Diplococcus pneumoniae, influenza, haemophilus etc. infect; Another kind of is the meningitis of mycobacterium tuberculosis infection.Clinical application is antibacterials associating hormone therapy mainly.Aseptic meningitis refers to that CSF smear and antibacterial culturing are recessive meningitis.General aseptic meningitis mostly is caused by viral infection.Viral encephalitis refers to the brain parenchyma inflammation caused by viral infection, often shows as heating, headache, tic, disturbance of consciousness and meningeal irritation symptom etc., can cause central nervous system's focal lesion.Viral encephalitis prognosis is not good, and mortality rate is high, often leaves serious sequela.Clinical commonly used drug is 1) hormones, as adrenocortical hormone dexamethasone; 2) antiviral purines, as acycloguanosine or vidarabine; 3) antiviral cell factor class, interferon; 4) nutrition class, as citicoline, vitamin B6, vitamin E, piracetam, pantothenic acid etc. improve brain metabolism, also in conjunction with antuepileptic, should can share ataraxy medicine etc. to the patient of psychomotor excitement to there being the patient of epilepsy.

The existing medicine for brain or nerve injury disease, still exists that side effect is large, weak curative effect, the defect such as cannot to effect a radical cure.Because brain diseases is a pathophysiological process relating to too many levels, multifactor, multipath complexity; and compound Chinese medicinal preparation can have many-sided neuroprotective; there is the clinical meaning of Mutiple Targets, multidirectional adjustment, control combination, should be expected to become the comparatively effective brain of one and nerve protection medicine.

Various Radix Ginseng is the good medicine of tonify deficiency regulation of mental activities, be often used in traditional Chinese medical science prescription, Shennong's Herbal records Radix Ginseng " main tonifying five ZANG-organs, peace spirit, determine soul, spasmolytic is throbbed with fear, except pathogen, ..., happy Fructus Alpiniae Oxyphyllae ", be considered to a kind of Chinese herbal medicine with multiple drug effect, its main pharmacodynamics active component is used for brain and many sections of reports have been shown in nerve injury, as senile dementia prevention and cure, parkinson, depression, cerebral infarction, (the rivers and mountains such as the other diseases that cerebral ischemia and cerebral ischemia cause, Jiang Zhenglin, Zeng Yinming, nine kinds of panaxoside monomer anti-cerebral ischemia damnification Effect study, Pharmacology and Clinics of Chinese Materia Medica 2007, 23 (2): 19-21, Xu slanders, Liu Chunqing, Ma Tao etc., and ginsenoside Re causes the protective effect of Parkinson disease mice dopaminergic neuron to MPTP, Shenyang Pharmaceutical University's journal, and 2005,22 (1): 36-44, Panaxsaponin composition and preparation method, patent of invention, application number 200610046083.3, compound 20S-ginsenoside Rh2 is preparing in antidepressant drug, patent of invention, application number 200810043537.0).

Rhizoma Coptidis property bitter cold, tool pathogenic fire purging, dampness, removing toxic substances, effect of parasite killing, modern pharmacology research shows, it is except having strong antibacterial activity, also has the multiple pharmacotoxicological effects such as antiinflammatory, blood fat reducing, blood sugar lowering, antitumor, antiulcer, nervous system protection.Report Rhizoma Coptidis is had in Cerebral ischemia and reperfusion damage, to play protective effect (Ki-Yeon Yool by antiinflammatory action; In Koo Hwang2; Jong Dai Kim3 etc.; Antiinflammatory Effect of the Ethanol Extract of Berberis koreana in a Gerbil Modelof Cerebral Ischemia/Reperfusion Phytother.Res.2008; 22,1527-1532).

Fructus Gardeniae is the fruit of Maguireothamnus speciosus Fructus Gardeniae (Gardenia jasminoides Ellis.), and property bitter cold is nontoxic.Enter the heart, liver, lung, stomach warp.There is effect of pathogenic fire purging relieving restlessness, clearing away heat and promoting diuresis, removing pathogenic heat from blood and toxic substance from the body.Cure mainly calentura restlessness of asrhenia type and insomnia, jaundice, gonorrhea, quench one's thirst, conjunctival congestion, pharyngalgia, haematemesis, epistaxis, dysentery, hematuria, pathopyretic ulcer, sprain and swell and ache.Clinical in diseases such as acute icterohepatitis, hemostasis, bruises.Modern pharmacology research shows, Fructus Gardeniae except having antipyretic, antibacterial, calm, analgesia, function of gallbladder promoting, antiinflammatory, stop blooding, protect the liver, except traditional pharmacological action such as soft tissue healing, also there is effect (chemical composition of Fructus Gardeniae and the pharmacological research such as antioxidation, asthma, blood fat reducing, blood pressure lowering, antiallergic, atherosclerosis, antiviral, antidepressant worry, nervous system protection, north pharmacy, 2008,5 (3): 44-45,51; Huang Shisun, Wu Shuyue, the modern pharmacological research of Fructus Gardeniae and clinical practice general introduction, internal medicine, 2010,5 (5): 534-536; Liu Guomin, Guo Suhua, Cheng Weiming, the pharmacological action of Fructus Gardeniae and Mechanism Study new development thereof, Strait Pharmaceutical Journal, 2008,20 (11): 8-10; Fructus Gardeniae extract is preparing the purposes prevented and/or treated in depressed worried medicine, Chinese invention patent, application number 200810016679.8).

The classical compound coptis chinensis Detoxication Decoction that Rhizoma Coptidis and Fructus Gardeniae share is published originally in " medical secrets of official ", is made up of, is regarded as the representative side of heat-clearing and toxic substances removing, have effect of eliminating fire and detoxication, cure mainly the card of all excess-heat fire-toxin, three burnt intenseness of heat Rhizoma Coptidis, Cortex Phellodendri, Radix Scutellariae, Fructus Gardeniae.There is report Rhizoma Coptidis toxic materials clearing away decoction for cerebropathy therapeutic, brain injury and the inflammatory that can reduce Cerebral ischemia and reperfusion invade profit (Young Sun Hwang a, Chan Young Shin b, YoungbuhmHuh etc., Hwangryun-Hae-Dok-tang (Huanglian-Jie-Du-Tang) extract and its constituentsreduce ischemia-reperfusion brain injury and neutrophil infiltration in rats.LifeSciences2002,71:2105-2117).

The present inventor notices still to lack and can be used for QI invigorating removing toxic substances with the pharmaceutical preparation for the treatment of brain and nerve injury disease.

Summary of the invention

The present composition derives from effective Chinese medicine compound QI invigorating Jiedu Fang of clinical treatment apoplexy.QI invigorating Jiedu Fang is the crucial pathogenesis for " poison damages brain network ", and employing determination for the treatment of based on pathogenesis obtained through differentiation of symptoms and signs, strengthening vital QI to eliminate pathogenic factors treatment traditional Chinese medical science encephalopathy propose.QI invigorating Jiedu Fang is the new compound Chinese medicinal preparation having no report in the past, and its active component is used for traditional Chinese medical science brain and nerve injury prevention and treatment of diseases also has no report.QI invigorating removing toxic substances prescription is made up of Radix Ginseng, Rhizoma Coptidis, Fructus Gardeniae three taste.Wherein, Radix Ginseng is monarch drug, strongly invigorating primordial QI, Fructus Alpiniae Oxyphyllae of calming the nerves, can tonifying five ZANG-organs gas, and the treatment deficient for healthy energy is very appropriate; Rhizoma Coptidis is ministerial drug, is the non-defective unit of eliminating fire and detoxication, and with ginseng compatibility, one mends one rushes down, and brings out the best in each other; Fructus Gardeniae is adjuvant, the kind relieving restlessness that clears away heart-fire of this medicine merit, and removing pathogenic heat from blood and toxic substance from the body, the eliminating fire and detoxication of auxiliary Rhizoma Coptidis, its property flexing is descending simultaneously, drives all pathogenic fire and goes out from urine, make the heresy outlet to some extent of pyretic toxicity.This three tastes compatibility, can on the basis of righting, and validation removing toxic substances is evil, plays good synergistic therapeutic action.

It is the general pathomechanism that brain injury is fallen ill that excitatory amino acid discharges in a large number with neurotransmitter exception, oxidative stress, free radical, inflammatory cascade, mitochondria dysfunction etc.; Radix Ginseng is longer than QI invigorating, neuroprotective; Rhizoma Coptidis and Fructus Gardeniae are longer than removing toxic substances, its three kinds of compatibilities can be accomplished reasonable compatibility, mutually short complementation.

Therefore, the technical problem to be solved in the present invention is to provide a kind of active ingredient of Chinese herbs compositions can preventing or treat brain and nerve injury disease.

The present invention relates to a kind of active ingredient of Chinese herbs compositions, described active ingredient of Chinese herbs comprises geniposide or jasminoidin, berberine, Radix Ginseng total saponins or ginsenoside monomer or ginsenoside monomer compositions.

As above according to compositions of the present invention, wherein said active ingredient of Chinese herbs is that 100 parts of calculating comprise following parts by weight proportioning according to gross weight: Radix Ginseng total saponins or ginsenoside monomer or ginsenoside monomer compositions, 1-99 part, berberine, 1-99 part, geniposide or jasminoidin, 1-99 part; Preferably ginseng's total saponins or ginsenoside monomer or ginsenoside monomer compositions, 2-75 part, berberine, 2-75 part, geniposide or jasminoidin, 2-75 part; More preferably Radix Ginseng total saponins or ginsenoside monomer or ginsenoside monomer compositions, 3-50 part, berberine, 3-50 part, geniposide or jasminoidin, 3-50 part.

As above according to compositions of the present invention, wherein the weight ratio of Radix Ginseng total saponins or ginsenoside monomer or ginsenoside monomer compositions, berberine, geniposide or jasminoidin is 3: 2: 2-0.1, preferably 3: 2: 2-1, more preferably 3: 2: 2.

As above according to compositions of the present invention, it comprises active ingredient of Chinese herbs and any one or multiple pharmaceutically acceptable carrier, such as antioxidant, chelating agent, filler, framework material etc.

As above according to compositions of the present invention, it can make the pharmaceutically acceptable dosage form of any one with pharmaceutically acceptable carrier according to specific needs, and such as tablet, granule, capsule, injection are as powder pin, freeze-dried powder, transfusion etc.

The invention further relates to compositions as above treat in preparation or prevent brain and nerve injury, such as cerebrovascular dementia or ischemic cerebrovascular (such as transience local cerebral ischemia outbreak, cerebral infarction or apoplexy), nervus retrogression encephalopathy, such as, purposes in the medicine of senile dementia or Parkinson's disease, bacillary or viral encephalitis or depression.

Accompanying drawing explanation

Fig. 1: the present composition to be performed the operation 24h cerebral Ischemia rat brain slice cerebral ischemia area TTC coloration result to row MCAO, A: sham operated rats; B: model group; C: positive controls (Folium Ginkgo extract Egb761 sheet group) (4mg/kg); D-F: administration group 3,2,1 (62.5mg/kg).

Fig. 2: in embodiment 1, the present composition is on the impact (n=10) of the rat cerebral infarction volume of row MCAO Post operation 12h and 24h.

Detailed description of the invention

Further illustrate the present invention below in conjunction with embodiment, advantage and disadvantage of the present invention will be more clear along with description.But these embodiments are only exemplary, do not form any restriction to scope of the present invention.It will be understood by those skilled in the art that and can modify to the details of technical solution of the present invention and form or replace down without departing from the spirit and scope of the present invention, but these amendments and replacement all fall within the scope of protection of the present invention.

Active ingredient of Chinese herbs preparation example

Berberine extraction process:

Get Rhizoma Coptidis, pulverize, put in apparatus,Soxhlet's, add 95% alcohol heating reflux to extracting liquid colourless, concentrating under reduced pressure 95% ethanol extract to paste, then adds 1% acetic acid, heating for dissolving, sucking filtration removing insoluble matter, drips concentrated hydrochloric acid in solution, to solutions turbid, place cooling, namely have yellow needles berberine hydrochloride to salt out, sucking filtration, washes twice crystal frozen water, then with washing with acetone once, accelerate dry, obtain crude product.By crude product water dissolution, regulate pH to be 8.5 ~ 9.8 with lime cream, cooling, filter out impurities, namely continue to be cooled to below room temperature has crystal to separate out, and sucking filtration, re-refines and get final product.

Stem and leaf of Radix Ginseng total saponins extraction process:

Get Stem and leaf of Radix Ginseng, be cut into 1 ~ 2cm section, decoct with water 2 times, 2 hours first times, second time 1.5 hours, merging filtrate, by D101 type macroporous resin, water elution is to colourless, use 60% ethanol elution again, collect 60% ethanol elution, eluent is concentrated into the clear paste that relative density is 1.06 ~ 1.08 (80 DEG C), drying and crushing, to obtain final product.

Geniposide or jasminoidin extraction process:

Get Fructus Gardeniae, pulverize, decoct with water 3 times, each 1.5 hours, decocting liquid filters, and merging filtrate, by AB-8 type macroporous resin, use water, 5% ethanol elution to colourless respectively, use 10% ethanol eluting again, collect 10% ethanol elution, eluent is concentrated into the clear paste that relative density is 1.06 ~ 1.08 (80 DEG C), drying and crushing, to obtain final product.

Ginsenoside monomer of the present invention can be ginsenoside Rg1, Rb1, Re, Rf1, Rd, Rh (Rh1 and Rh2).

In this patent, ginsenoside monomer includes but not limited to be obtained by following method:

(I) preparation of ginsenoside monomer Rf1 is prepared (medical usage of GF1, application number 200610008477.X) with reference to patented method;

(II) preparation of ginsenoside monomer Re is prepared (ginsenoside Re's new medical use and preparation method, application number 2004110021133.6) with reference to patented method;

(III) preparation of ginsenoside monomer Rg1 is prepared (purposes of ginsenoside Rg1, application number 200810227643.4) with reference to patented method;

(IV) preparation of Panaxadiol Saponins Monomer Rb1 is prepared (preparation method of a kind of ginsenoside Rb1, application number 200710178212.9) with reference to patented method;

(V) preparation of ginsenoside monomer Rh (Rh1 and Rh2) is prepared (ginsenoside Rh's extract and preparation method, application number 200810043537.0) with reference to patented method;

(VI) the reference literature method of preparing of ginsenoside monomer Rd is prepared (Liu Deyu, Cao Sa.The method of rapid batch separation and Extraction ginsenoside Rd from pseudo-ginseng, Chinese crude drug, 2006,29 (3): 247-249).

The above Radix Ginseng total saponins and ginseng monomer are the known material with pharmaceutically active, by Radix Ginseng total saponins or above-mentioned various ginsenoside monomer or two or more ginsenoside monomers mix can be used as the crude drug of pharmaceutical preparation after remix by following certain proportion with berberine, jasminoidin, formulation method routinely, be prepared into clinical applicable any one medicament, the oral solid formulations such as such as tablet, capsule, granule, drop pill, oral liquid, injection etc.In the middle of the process of the above-mentioned preparation of preparation, the mixture of above-mentioned preparation and any one applicable mixed with excipients for the preparation of clinical medicament can be used, be prepared into pharmaceutical preparation.

Pharmaceutical composition matching method

(1) calculate according to parts by weight proportioning, get respectively prepare according to the method described above Radix Ginseng total saponins 30 parts, after berberine 20 parts and the mixing of jasminoidin 5-20 part as the crude drug of pharmaceutical preparation;

(2) calculate according to parts by weight proportioning, get respectively prepare according to the method described above any one ginsenoside monomer 30 parts, after berberine 20 parts and the mixing of jasminoidin 5-20 part as the crude drug of pharmaceutical preparation;

(3) calculate according to parts by weight proportioning, get respectively prepare according to the method described above 30 parts, two or more ginsenoside monomer mixture, after berberine 20 parts and the mixing of jasminoidin 5-20 part as the crude drug of pharmaceutical preparation.

Pharmaceutical composition of the present invention share according to as above (1)-(3) matching method composition compositions and berberine and jasminoidin and forms with pharmaceutically acceptable excipient or carrier.

By add in any one compound shown in above-mentioned (I)-(VI) of the present invention prepare different dosage form time required various adjuvant or pharmaceutically acceptable excipient or carrier after, can be prepared into the suitable clinical preparation of any one with the drug formulation process of routine, such as, can be injection (powder pin, freeze-dried powder, liquid drugs injection, transfusion etc.), tablet, oral liquid, granule, capsule, soft capsule, drop pill etc.Wherein, described adjuvant can be antioxygen chelating agent, filler, framework material etc.; Described pharmaceutically acceptable carrier is one or more in xylitol, mannitol, lactose, fructose, glucosan, glucose, polyvinylpyrrolidone, low molecular dextran, sodium chloride, calcium gluconate or calcium phosphate, is preferably mannitol or lactose.

Embodiment 1 present composition is tested acute focal cerebral ischemia animal protection Effect study

Acute focal cerebral ischemia in rats sets up (row middle cerebral artery thromboembolism (MCAO) operation)

Adopt body weight at 250-270g male SD rat, with reference to (Longa EZ such as Zea Longa, Weinstein PR, Carlson S, Cummins R.Reversible middle cerebral artery occlusion withoutcraniectomy in rats.Stroke1989; Bolt line blocking-up method in intraluminal middle cerebral artery occlusion in rats 20:84-91) set up.10% chloral hydrate solution 400mg/kg, intraperitoneal injection of anesthesia animal.Rat dorsal position is fixed, cervical region medisection skin, each layer tissue of blunt separation, exposes right carotid (CCA).To be separated to after internal carotid artery (internalcarotid artery, ICA), external carotid artery (external carotid artery, ECA) bifurcated one section, to be carefully separated and to avoid damage vagus nerve and trachea, put line for subsequent use.Be separated homonymy external carotid artery, send about 0.8cm place's ligation at ECA.A bulldog clamp is pressed from both sides in CCA proximal part, " V " shape otch that a diameter is about 2mm is done between ECA ligation place and crotch, nylon wire is inserted CCA gently from incision, unclamp bulldog clamp, in neck, external carotid artery furcation enters internal carotid artery, the direction that nylon wire enters intracranial to ICA is lentamente advanced, insertion depth about 18.5 ± 0.5mm is to micro-sense resistance, make nylon wire head end by MCA section start, arrive thinner anterior cerebral artery, now namely realize the blood flow obstruction of left side middle cerebral artery, ligation ICA is with fixing nylon wire and prevent hemorrhage, layer-by-layer suture, nylon wire stump stays 1cm to be longer than outside skin.

Sham operated rats only carries out preoperative anesthesia and blood vessel exclusion, not ligation and lead-in wire bolt.In operation process, room temperature remains on 24-25 DEG C, and biological functional system carries out animal breath and cardioelectric monitor.

Animal grouping and administration

SD rat is divided into six groups at random: sham operated rats, model group, Folium Ginkgo extract sheet positive controls (Ginaton Folium Ginkgo extract sheet, Egb761, Dr Willmar Schwabe), present composition administration group 1,2,3.Positive controls medication: get Folium Ginkgo extract sheet 1 (40mg) and be dissolved in 40ml normal saline, be mixed with 1mg/mL, gavage gives SD rat 1mL, and dosage is 4mg/kg.Administration group medication: get Radix Ginseng total saponins respectively: berberine: jasminoidin is according to weight ratio 3: 2: 2 (administration group 1), 3: 2: 0.1 (administration groups 2), 3: 2: 0.5 (administration groups 3), 31.25mg/mL is become respectively with normal saline, gastric infusion 0.5mL, dosage is 62.5mg/kg.Administration group and positive controls be 10min gastric infusion before surgery anesthesia, respectively at postoperative 4,12,24h puts to death animal, carries out following indices detection.

The functional evaluation of neuroethology Bederson ' s method

Animal carries out neuroethology observation before execution, with reference to method (the list of references Bederson JB of Bederson etc., Pitts LH, Tsuji M, et al., Rat middle cerebral artery occlusion:evaluationof the model and development of a neurologic examination.Stroke1986; 17:472-476), carry Mus tail and leave ground about 1 chi, observe two forelimb situations; Rat is placed in level ground, promotes its both shoulders, observing both sides resistance has zero difference; Rat is placed in ground, observes its walking situation.Adopt Pyatyi point system (0-4 divides), mark is higher, illustrates that the damage of its neurobehavioral is more serious.The statistical result of Bederson ' s function score is in table 1, and between each group, data compare employing rank test, and P < 0.05 thinks to have statistical significance.

The statistical result of table 1Bederson ' s function score

Note: nonparametric rank test, inspection, compare with matched group: ^*p < 0.01; Compare with model group: #P < 0.05, ##P < 0.01

Conclusion: as shown in Table 1, model group comparatively sham operated rats has significant difference, modeling success (P < 0.01) is described, positive controls (3mg/kg) and administration group 1,2,3 significantly can improve the nerve injury symptom (P < 0.05, P < 0.01) of 12h and 24h after animal focal cerebral ischemia.

To the improvement result evaluation of cerebral ischemia cerebral infarction volume

Rat sacrificed by decapitation after function score, is placed in-20 DEG C of refrigerators by the cerebral tissue of taking-up rapidly, the rearmounted room temperature environment of 10min, excision olfactory bulb, cerebellum and low brain stem after and be cut into 6 brain continuous coronal rough lumber sheets by interval 2mm shown in cerebral localization collection of illustrative plates is crown.Then the rapid 5ml that is placed in by brain sheet is containing 2%TTC (triphenyltetrazolium chloride) and 1molL ^-1k ₂hPO ₄in the solution of 0.1ml, 37 DEG C of constant temperature, lucifuges hatch 30min, and brain sheet stirs once every 5min by period.After TTC dyeing, normal structure is rose, and infarction tissue is not colored and is white.Often will organize brain sheet marshalling, preservation of taking pictures, the process of applies image analysis systems soft ware also takes statistics, and calculates the infarct size of often opening brain sheet, is multiplied by the thickness 2mm of every sheet brain sheet, and every animal all brain sheets infarct size is multiplied by thickness and is added, and is cerebral infarct volume.

Conclusion: as illustrated in fig. 1 and 2, after rat cerebral tissue's ischemic injuries, sham operated rats occurs without cerebral infarction, and model group occurs obvious infarcted region and presents TTC dyeing canescence.Administration group 1,2,3 significantly can reduce cerebral infarction ash white area, reduces cerebral infarct volume, has remarkable improvement result to cerebral infarction.

To the improvement result evaluation of local cerebral blood flow after middle cerebral artery thromboembolism (MCAO) rat cerebral ischemia

MCAO operation method is described above, and medicine 15min before chloral hydrate surgery anesthesia gives.

Animal grouping and medication

SD rat is divided into following several groups at random: sham operated rats, model group, Folium Ginkgo extract sheet positive controls (Ginaton Folium Ginkgo extract sheet, Egb761, Dr Willmar Schwabe), present composition administration group (Radix Ginseng total saponins: berberine: jasminoidin is according to weight ratio 3: 2: 2).Positive controls medication: get Folium Ginkgo extract sheet 1 (40mg) and be dissolved in 40ml normal saline, be mixed with 1mg/mL, gavage gives SD rat 1mL, and dosage is 4mg/kg.Administration group medication: administration group becomes 0.25mg/mL, 1.25mg/mL and 6.25mg/mL with normal saline respectively according to proportion compatibility, gastric infusion 1mL, three dosage are respectively 1mg/kg, 5mg/kg and 25mg/kg from high to low.Supine is positioned on operating-table, anaesthetize but do not carry out MCAO operation consent, after stabilised blood purling fluence reading, laser speckle blood flow monitoring video system (PariCam PSI) is used to record Blood Flow Value in 15min, the basic value (baselinevalue) using its meansigma methods as cerebral blood flow.Complete MCAO operation, line is inserted intracranial, model group is when Blood Flow Value drops to suddenly the 40%-60% of basic value, prompting mesencephalic arteries blood flow is blocked, same model group is respectively organized in administration, the preoperative Blood Flow Value of row MCAO is the basic value (100%) of this group, and Post operation Blood Flow Value all represents with the percentage rate of this basic value.Post operation 30min measures LDF value at same position, and the change of monitoring SD rats underwent MCAO perioperatively blood flow, this system not only can grasp the spatial distribution of blood perfusion, and clearly can see the situation of blood flow Real-time and Dynamic change clearly.The present composition to SD rats underwent MCAO postoperative brain blood flow exercising result in table 2.

The computational methods of average blood flow groundwater increment (PU) are:

Average blood flow groundwater increment (PU)=hemocyte translational speed (V) × hematocrite concentration (C)

Utilize following formulae discovery local cerebral blood flow rate of change:

Wherein, TOI _controlfor auricle average blood flow groundwater increment (operation consent) before administration, TOI _treatedfor auricle average blood flow groundwater increment (Post operation) after administration.

Table 2 present composition is to SD rats underwent MCAO postoperative brain blood flow effect (n=5)

Note: nonparametric rank test, inspection, compare with matched group: ^*p < 0.01; Compare with model group: ^##p < 0.01

Conclusion: by regional cerebral blood flow amount (rCBF) change in half an hour after monitoring ischemia, the local cerebral blood flow reduced after positive drug group (4mg/kg) and the high, medium and low dosage of administration group (1mg/kg, 5mg/kg, 25mg/kg) can obviously improve focal cerebral ischemia in row MCAO Post operation ischemia half an hour, and present composition administration group low dose group (1mg/kg, 32.96 ± 5.17 ^*) still show and improve regional flow's groundwater increment effect preferably, compared with model group, tool significant difference (P < 0.01), and present concentration-dependent relation well.This shows present composition energy fast strikethrough blood brain barrier and by blood vessel dilating, can increase regional flow's groundwater increment and play good vascular protection effect.

By testing the known present composition above, there is good Acute cerebral ischemia protective effect.

Embodiment 2 present composition is to focal cerebral ischemia animal postischemic reperfusion protective effect research experiment

The preparation of middle cerebral artery thromboembolism (MCAO) postischemic reperfusion model

Adopt body weight at 250-270g male SD rat, with reference to (Longa EZ such as Zea Longa, Weinstein PR, Carlson S, Cummins R.Reversible middle cerebral artery occlusion withoutcraniectomy in rats.Stroke1989; In intraluminal middle cerebral artery occlusion in rats 20:84-91) set up, bolt line blocking-up method, suitably improves.

10% chloral hydrate solution 400mg/kg, intraperitoneal injection of anesthesia animal.Rat dorsal position is fixed, cervical region medisection skin, each layer tissue of blunt separation, exposes right carotid (CCA).To be separated to after internal carotid artery (internalcarotid artery, ICA), external carotid artery (external carotid artery, ECA) bifurcated one section, to be carefully separated and to avoid damage vagus nerve and trachea, put line for subsequent use.Be separated homonymy external carotid artery, send about 0.8cm place's ligation at ECA.A bulldog clamp is pressed from both sides in CCA proximal part, " V " shape otch that a diameter is about 2mm is done between ECA ligation place and crotch, nylon wire is inserted CCA gently from incision, unclamp bulldog clamp, in neck, external carotid artery furcation enters internal carotid artery, the direction that nylon wire enters intracranial to ICA is lentamente advanced, insertion depth about 18.5 ± 0.5mm is to micro-sense resistance, make nylon wire head end by MCA section start, arrive thinner anterior cerebral artery, now namely realize the blood flow obstruction of left side middle cerebral artery, ligation ICA is with fixing nylon wire and prevent hemorrhage, layer-by-layer suture, nylon wire stump stays 1cm to be longer than outside skin.

Sham operated rats only carries out preoperative anesthesia and blood vessel exclusion, not ligation and lead-in wire bolt.In operation process, room temperature remains on 24-25 DEG C, and biological functional system carries out animal breath and cardioelectric monitor.

Animal grouping and administration

SD rat is divided into following several groups at random: sham operated rats, model group, Folium Ginkgo extract sheet positive controls (Ginaton Folium Ginkgo extract sheet, Egb761, Dr Willmar Schwabe) (4mg/kg), present composition administration group (Radix Ginseng total saponins: berberine: jasminoidin is according to weight ratio 3: 2: 2).Positive controls medication: get Folium Ginkgo extract sheet 1 (40mg) and be dissolved in 40ml normal saline, be mixed with 1mg/mL, give SD rat 1mL in Post operation 3h gavage, dosage is 4mg/kg.Administration group medication: administration group becomes 0.25mg/mL, 1.25mg/mL and 6.25mg/mL with normal saline respectively according to proportion compatibility, in Post operation 3h gastric infusion 1mL, three dosage are respectively 1mg/kg, 5mg/kg and 25mg/kg from high to low.Except sham operated rats, all the other each group after ischemia 2h residual fishing line to be extracted to neck and external carotid artery crotch, cause reperfusion injury, after 12h and 24h, put to death animal carry out index of correlation detection.

The functional evaluation of neuroethology Bederson ' s method

Animal carries out neuroethology observation before execution, with reference to method (the list of references BedersonJB of Bederson etc., Pitts LH, Tsuji M, et al., Rat middle cerebral artery occlusion:evaluation of themodel and development ofa neurologic examination.Stroke1986; 17:472-476), carry Mus tail and leave ground about 1 chi, observe two forelimb situations; Rat is placed in level ground, promotes its both shoulders, observing both sides resistance has zero difference; Rat is placed in ground, observes its walking situation.Adopt Pyatyi point system (0-4 divides), mark is higher, illustrates that the damage of its neurobehavioral is more serious.

Appraisal result is learned in table 3 by function of nervous system, and between each group, data compare employing rank test, and P < 0.05 thinks to have statistical significance.

Table 3 present composition is on the impact of rat function assessment scoring after cerebral ischemia reperfusion

Data are represented with meansigma methods ± standard deviation and are analyzed by one-way ANOVA.Relative to sham operated rats: ^##p < 0.01, ^#p < 0.05; Relative to model group: ^*p < 0.05, ^*p < 0.01.

Conclusion: model group comparatively sham operated rats has significant difference, modeling success (P < 0.01) is described, positive control Folium Ginkgo extract group (4mg/kg, ig) and the present composition large, medium and small dosage group (1,5mg/kg, 25mg/kg) significantly can improve nerve injury symptom (the P < 0.05 that animal focal cerebral ischemia fills with rear 12h and 24h again, P < 0.01), and present certain dose-dependence.

To the improvement result evaluation of cerebral ischemia cerebral infarction volume

Rat sacrificed by decapitation after function score, is placed in-20 DEG C of refrigerators by the cerebral tissue of taking-up rapidly, the rearmounted room temperature environment of 10min, excision olfactory bulb, cerebellum and low brain stem after and be cut into 6 brain continuous coronal rough lumber sheets by interval 2mm shown in cerebral localization collection of illustrative plates is crown.Then the rapid 5ml that is placed in by brain sheet is containing 2%TTC (triphenyltetrazolium chloride) and 1molL ^-1k ₂hPO ₄in the solution of 0.1ml, 37 DEG C of constant temperature, lucifuges hatch 30min, and brain sheet stirs once every 5min by period.After TTC dyeing, normal structure is rose, and infarction tissue is not colored and is white.Often will organize brain sheet marshalling, preservation of taking pictures, the process of applies image analysis systems soft ware also takes statistics, and calculates the infarct size of often opening brain sheet, is multiplied by the thickness 2mm of every sheet brain sheet, and every animal all brain sheets infarct size is multiplied by thickness and is added, and is cerebral infarct volume.The present composition for the effect of cerebral ischemia reperfusion cerebral infarction volume in table 4.

Table 4 present composition is to the effect of cerebral ischemia reperfusion cerebral infarction volume

Data are represented with meansigma methods ± standard deviation and are analyzed by one-way ANOVA.For sham operated rats ^##p < 0.01, ^#p < 0.05; For model group ^*p < 0.05, ^*p < 0.01.

Conclusion: it is as shown in the table, after rat cerebral tissue's ischemical reperfusion injury, sham operated rats occurs without cerebral infarction, and obvious infarcted region appears in model group.Administration group (1,5mg/kg and 25mg/kg) significantly can reduce cerebral infarct volume.

Cerebral edema improvement result is evaluated

Cerebral edema is, with cerebral ischemia, pathological process important in early days occurs, and evaluating the improvement result of medicine to cerebral edema is the important indicator evaluating medical treatment effect, adopts brain water content and cerebral edema rate to evaluate.

Concrete Testing index:

Brain water content (%)

Ischemia side and sinistrocerebral weight in wet base is taken immediately after getting brain.Then take dry weight after left hemisphere being put the oven for baking 24h of 100 DEG C, calculate brain water content.Brain water content (%)=(cutaneous horn weight-brain stem weight)/cutaneous horn heavy × 100%.

The present composition on the impact of rat cerebral tissue's water content after cerebral ischemia reperfusion in table 5.

Table 5 present composition is on the impact of rat cerebral tissue's water content after cerebral ischemia reperfusion

Data are represented with meansigma methods ± standard deviation and are analyzed by one-way ANOVA.For sham operated rats ^##p < 0.01, ^#p < 0.05; For model group ^*p < 0.05, ^*p < 0.01.

Conclusion: cerebral ischemia reperfusion 12h and 24h, model group is compared with matched group, cerebral edema rate all has significant difference (P < 0.05), positive controls has significant difference (P < 0.05 with each dosage group of the present composition compared with model group, P < 0.01), illustrate that the present composition obviously can improve the cerebral edema of cerebral ischemia 12 and 24h rat.

In sum; the improvement of scoring and cerebral edema is learned from function of nervous system; the minimizing of cerebral infarction volume, shows the effect damaged after the large, medium and small dosage group of the present composition (1,5mg/kg and 25mg/kg) has obvious protection Cerebral Ischemia-reperfusion.

Embodiment 3 present composition is tested senile dementia (AD) animal protection Effect study

Amyloid-beta (A β _1-40) hatch

With physiological saline solution by A β _1-40be diluted to 2ug/ul, hatch 1 week for 37 DEG C, make it the A β becoming condensed state _1-40.

A β _1-40cause Model of Dementia in Rats to set up

Be fixed on rat stereotaxic instrument after getting the healthy male AD rat anesthesia of 250-270g, conventional preserved skin sterilization, cut skin, expose bregma, slowly condensed state A β is injected to rats with left hippocampus (be 0 point with Bregma point, AP-3.0mm, ML2.0mm, DV-2.9mm) with microsyringe _1-405 μ l, let the acupuncture needle remain at a certain point 5min, to make the abundant disperse of A β, then slowly remove pin, sew up the incision.Sham operated rats only carries out preoperative anesthesia, cuts skin, exposes bregma, does not carry out injection A β _1-40.

Experiment grouping and administration

Healthy for 250-270g male SD rat 30 is divided into sham operated rats, A β model group, Folium Ginkgo extract positive controls, present composition administration group 1,2,3 at random, often organizes 5.Positive controls medication: get Folium Ginkgo extract sheet 1 (40mg) and be dissolved in 4ml normal saline, be mixed with 1mg/mL, gavage gives SD rat 1mL, and dosage is 4mg/kg.Administration group medication: get Radix Ginseng total saponins respectively: berberine: jasminoidin is according to weight ratio 3: 2: 0.5,62.5mg/mL is become with normal saline, administration group 1,2,3 respectively gavage gives 2mL, 1mL and 0.5mL, and namely dosage is 500mg/kg, 250mg/kg and 125mg/kg.After model has been made, A β model group has poured into equal-volume normal saline, and other groups pour into corresponding medicine.

Morris water maze is tested

Postoperative 30d starts determined with Morris water.Water maze is diameter 90cm, the pond of high 40cm, depth of water 30cm, pool wall is indicated 4 place of entry, is divided into four quadrants, put a lucite circular platform in a wherein quadrant center, platform is lower than water surface 2cm, water temperature (24 ± 2) DEG C, by the uniform white plastic foam coverage of opaque thickness, peripheral pool object of reference remains unchanged.Test comprises:

1) orientation navigation test: for measuring the acquisition capability of rat to water fan official learning and memory.Last 5d, two time periods of morning and afternoon every day, each time period 4 times, respectively from four different gauge points, rat is put into water towards pool wall, in record 2min, find platform required time (escape latency).If fail to find platform in 2min after rat enters water, to be then placed on platform and to stop 10s, escape latency record 2min.Each training interval 60s.

2) space exploration test: after measuring rat association searching platform, to the hold facility of platform space position memory.After orientation navigation off-test, removed by platform in the 6th day, rat is put into water towards pool wall by an optional place of entry, crosses over the number of times of original platform position in record 2min.

Adopt one factor analysis of variance (One-Way ANOVA), P < 0.05 thinks to have statistical significance.Morris water maze test result is in table 6.

Average escape latency in the test of table 6 orientation navigation

Note: ^*p < 0.01, #P < 0.05, ##P < 0.01

Table 7, Morris water maze rat space exploration experimental result

Note: ^*p < 0.05, ^*p < 0.01

As shown in Table 6, rat changes not obvious in after giving the present composition first 2 days, and from the 3rd day, model group was compared with sham operated rats: ^*p < 0.01, illustrates modeling success; Administration group compares with A β model group: ^#p < 0.05, ^##p < 0.01, this shows, administration group 1,2,3 obviously can shorten the latter three days average escape latency started.Table 7 shows, administration group obviously can shorten rat space exploration incubation period, and increase and cross over original platform position number of times, therefore the present composition is to A β _1-40caused rat AD model has obvious protective effect.

Aluminium oxide causes mice Model of Dementia and medication

Laboratory animal selects the aged Kunming mouse at 22 monthly ages, and male and female half and half are divided into 6 groups at random: Normal group, aluminum chloride model group, aluminum chloride add positive control Folium Ginkgo extract sheet group, aluminum chloride adds present composition administration group 1,2,3.Positive controls administration compound method: get Folium Ginkgo extract sheet 1 (40mg) and be dissolved in 4ml normal saline, be mixed with 1mg/mL, gavage gives SD rat 1mL, and dosage is 4mg/kg.Administration group administration compound method: get Radix Ginseng total saponins respectively: berberine: jasminoidin is according to weight ratio 3: 2: 0.5,62.5mg/mL is become with normal saline, administration group 1,2,3 respectively gavage gives 2mL, 1mL and 0.5mL, namely dosage is 500mg/kg, 250mg/kg and 125mg/kg.To aluminum chloride distilled water solution (Shanghai Jinshan chemical plant produces, lot number 100093) the 200mg/kg body weight/day of aluminum chloride model group mice 10mg/ml, gastric infusion, continuous 90 days.While setting up Model of Dementia, Folium Ginkgo extract sheet and the present composition is given, gastric infusion, continuous 90 days to positive controls and administration group.Normal group gives same dosage normal saline.

Jumping test

TT-2 type mice diving tower procedure auto-control instrument (institute of Materia Medica,Chinese Academy of Medical Sciences development).Its diving tower reflective box is 90 × 18 × 60cm ³lucite case, is separated between 5 by lamella lucida, is copper grid at the bottom of case, in addition 36V voltage, a built-in high 3.5cm between every, and diameter is the valve rubber of 3.5cm, avoids the place of safety of electric shock as mice.Test front 3 angel mices and be familiar with laboratory environment, keep quite in indoor, temperature 25 DEG C.After mice being put into reflective box endoadaptation 5min during experiment, then pass to 0.25mA electric current, be placed in by mice on diving tower, mice has the habit of jumping off from eminence, if jumped off, is shocked by electricity and produces memory.During training, diving tower is greater than 180s person is discarded incubation period, the number of times (errors number) jumped off from platform in record mice 5min, based on errors number, mice is divided into 6 groups of (Normal group, model group, positive control Folium Ginkgo extraction sheet group, administration groups 1,2 by achievement, 3, positive control and administration 1,2,3 groups), often organize 10,6 groups of intelligent of ageing mice are on close level.Conscientiously measure mice in 5min during experiment in detail and jump off the number of times that contacts to earth (error number) from platform, learn the observation index of acquisition and memory consolidation as mice.

Adopt one factor analysis of variance (One-Way ANOVA) analysis, P < 0.05 thinks to have statistical significance.Jumping test the results are shown in Table 8.

The comparison of table 8 mice Jumping test ability of learning and memory change

Note: ^*p < 0.05, #P < 0.05, ##P < 0.01

As shown in Table 8, model group compares with administration group: ^*p < 0.05, illustrates modeling success; Administration group compares with model group: ^#p < 0.05, ^##p < 0.01, shows that the present composition obviously can reduce the errors number of mice Model of Dementia, is therefore significantly improved effect to ability of learning and memory in mice.

Embodiment 4 present composition is to parkinsonian animal models protective effect research experiment

Parkinsonian animal models is set up and medication

MPTP-induced Parkinson disease mice model is made by the method for lumbar injection pesticide N,N'-dimethyl-.gamma..gamma.'-dipyridylium (paraquat, PQ).Get Kunming mouse 120 in male 7 week age, after adapting to one week, be divided into 6 groups at random, often organize 20.Blank group: gavage distilled water 10mL/kg body weight once a day, weekly intraperitoneal injection of saline 10ml/kg body weight; All the other 5 groups weekly lumbar injection PQ10mg/kg, and gavage every day colchicine, by 4 times of adult's dosage, i.e. 0.2mg/kg, continues modeling administration in 7 weeks.After modeling success, positive controls: gavage every day madopar, according to adult 7.5 times of dosage, namely presses 125mg/kg dosed administration, to continue medication to modeling one week; Administration group 1,2,3: respectively mouse stomach present composition 250mg/kg, 125mg/kg, small dose group 62.5mg/kg (compound method is with embodiment 3), to continue medication to modeling one week.Raise under the same terms, the behavioristics observing mice changes, and then does the content detection dopamine (DA) of mice black substance brain district monoamine neurotransmitter, norepinephrine (NE), 5-hydroxy tryptamine (5-HT).

Turn-club test

Normal mouse has climbing ability, and this needs suitable grip and sports coordination, and we adopt turn-club test to detect the dyskinesia of mice.Mice is put on transfer rod tester (Ji'an, Beijing get Er Science and Technology Ltd. YLS-4C type), puts 6 at every turn, rotate by the speed of 20 turns per minute, the number of times fallen down in record mice 3min.

Autonomic activities measures

Mice is put into the active box of autonomic activities tester, each Simultaneously test 6 mices, 1 mice in each case, puts into relief mice adaptation 3min and starts test again, after computer enters mensuration program, automatically record number of times that is movable in mice 10min and that stand by computer.

Adopt one factor analysis of variance (One-Way ANOVA) analysis, P < 0.05 thinks to have statistical significance.Behavioristics's testing result is in table 9.

Table 9 behavioristics testing result

Note: ^*p < 0.01, ^#p < 0.05, ^##p < 0.01

As shown in Table 9, the number of times that after giving the present composition, mice falls down from transfer rod reduces, and movable number of times obviously increases.Administration group compares with model group: ^#p < 0.05, ^##p < 0.01, illustrates that the present composition significantly can improve the behavioral competence of parkinson mice.

The content detection of monoamine neurotransmitter the results are shown in Table 10.

The measurement result of table 10DA, NE, 5-HT content

Note: ^*p < 0.05, ^*p < 0.01, ^#p < 0.05, ^##p < 0.01

From table 9,10, the present composition significantly can reduce the dyskinesia that PQ causes parkinson mice, reduces the excessive release of monoamine neurotransmitter, improves parkinson symptom.

Embodiment 5 present composition is to creation protective effect research experiment

Animal Model

Adopt permanent ligation bilateral common carotid arteries rat models of vascular dementia.After SD rats by intraperitoneal injection 3.5% chloral hydrate 10ml/kg anaesthetizes, dorsal position is fixed, cervical region median incision, is separated bilateral common carotid arteries, avoids damage cervical sympathetic nerve and vagus nerve, and with Line 1 by its permanent ligation, postoperative sew up wound, raises in cage.Sham operated rats is for being separated bilateral common carotid arteries, but not ligation.

Animals administer method

Operation January after, grouping administration, rat is divided into six groups at random: sham operated rats, model group, Folium Ginkgo extract sheet positive controls (4mg/kg), present composition administration group 1,2,3 (250mg/kg, 125mg/kg, 62.5mg/kg) (compound method is with embodiment 3).

Morris water maze ability of learning and memory test experience

Water maze is diameter 90cm, the pond of high 40cm, depth of water 30cm, pool wall is indicated 4 place of entry, is divided into four quadrants, put a lucite circular platform in a wherein quadrant center, platform is lower than water surface 2cm, water temperature (24 ± 2) DEG C, by the uniform white plastic foam coverage of opaque thickness, peripheral pool object of reference remains unchanged.Test comprises:

1) course experiment is located: for measuring the acquisition capability of rat to water fan official learning and memory.Platform is placed in the centre of a certain quadrant in labyrinth, an optional place of entry in other three quadrants, every day is trained secondary in experiment, the place of entry that each employing is different, each training scheme of 1 minute.If animal not yet finds platform within 2min, then being taken by animal on platform and to make it stand above 15 seconds, if rat found platform within 2 minutes, be also allowed to condition on platform and stand 15 seconds, reef knot Shu Yici trains.Training like this five days, records escape latency and the swimming path of every animal respectively.

2) space search experiment: after measuring rat association searching platform, to the hold facility of platform space position memory.After orientation navigation off-test, in the 6th day, platform is removed, every rat only swims 2 minutes, respectively with the time of stopwatch record every rat first time through platform, i.e. incubation period, and record every rat number of times through platform in 2 minutes, if the incubation period of certain rat is shorter and more through original platform position number of times, then illustrate this rat space orientation ability and learning and memory function good.

Administration started Morris water maze laboratory after 14 days.Located course experiment before this, every rat trains 2 times every day, and training duration is 120s, continuously training 5 days, record parameters.Then carry out space search experiment, within the 6th day, remove platform, record rat incubation period and in 2 minutes by the number of times of position of platform.

Adopt one factor analysis of variance (One-WayANOVA) analysis, P < 0.05 thinks to have statistical significance.Water maze laboratory the results are shown in Table 11,12,13,14.

Table 11 vascular dementia rats location course experiment escape latency

Note: ^*p < 0.05, ^*p < 0.01, ^##p < 0.01

As shown in Table 11, model group compares with sham operated rats, ^#p < 0.05, ^##p < 0.01, modeling success is described, administration group compares with model group, ^*p < 0.05, ^*p < 0.01, illustrates that the present composition significantly can reduce the escape latency of vascular dementia rats.

The table 12 vascular dementia rats location course experiment distance of swimming

Note: ^*p < 0.05, ^*p < 0.01, ^##p < 0.01

Have table 12 known, model group compares with sham operated rats, ^#p < 0.05, ^##p < 0.01, modeling success is described, administration group compares with model group, ^*p < 0.05, ^*p < 0.01, illustrates that the present composition significantly can reduce the distance of swimming in water maze laboratory of vascular dementia rats.

Table 13 vascular dementia rats space exploration experimental result

Note: ^*p < 0.05, ^*p < 0.01

As shown in Table 13, administration group compares with model group, ^*p < 0.05, ^*p < 0.01, illustrate incubation period that rat gives present composition rear space explorative experiment comparatively model group obviously shorten, obviously increase through original platform number of times.

The table 14 different dosing natural law present composition finds the impact of platform experiment incubation period on intending VD rat

Note: ^*p < 0.05, ^*p < 0.01, ^##p < 0.01

As shown in Table 14, administration group compares with model group, ^##p < 0.01, illustrates the time that can shorten after rat gives the present composition and find platform.

From above experimental result, after giving the present composition, obviously can improve the ability of learning and memory of vascular dementia rats.

Step-through test

Rat is put into bright room, free movable 3min in case.Do not enter darkroom person in 3min and reject (during preliminary election no power).Formally carry out learning training after 1h, it is incubation period that record rat enters the darkroom required time that gets shocked, and records in 5min and suffer number of shocks (errors number) as school grade; Repeat said process after 24h, record incubation period and errors number are as Memory result.Voltage is 36V, incubation period index with second (S) for unit.

Adopt one factor analysis of variance (One-WayANOVA) analysis, P < 0.05 thinks to have statistical significance.Step-through test the results are shown in Table 15.

Table 15 vascular dementia rats step-through test result

Note: ^*p < 0.01, ^##p < 0.01

As shown in Table 15, administration group 1 compares with model group ^##p < 0.01, illustrate that the present composition obviously can shorten the incubation period of learning training and recall tests, and reduce errors number, the visible present composition can improve the ability of learning and memory of vascular dementia rats.

Conclusion: the present composition significantly can improve the ability of learning and memory that permanent ligation bilateral common carotid arteries makes vascular dementia rats, therefore has protective effect to vascular dementia rats.

Embodiment 6 present composition is tested stress depression animal protection Effect study

First letting animals feed 1 week in quiet, warm environment before experiment, all single cage raising except matched group, natural lighting, well-ventilated, ad lib water.

Wistar rat chronic is slight Unpredictability stress (CUMS) depression model production program: 2h behavior is limited, 4 DEG C of frozen water went swimming 5min, the 24h that cut off the water, food 24h, folder tail 1min, light and shades puts upside down 24h, electric shock vola 5s (30V voltage), rock 5min (160Hz), 40 DEG C of environment 5min etc. nine kinds stimulates random arrangement in 18 days, every day is a kind of, makes animal can not expect the generation stimulated.Be divided into by Wistar rat Normal group, depression model group, fluoxetine positive controls, administration group 1,2,3 (250mg/kg group, 125mg/kg group, 62.5mg/kg group) (compound method is with embodiment 3) to continue 21 days at random, each treated animal is sacrificed by decapitation after last administration 24h.

Experimental index measures

Behavioristics measures---and Morris water maze is tested

The training and testing of spatial memory carries out with reference to Morris water Maze method.Water temperature remains on about 23 DEG C, and water is dyed to white, with hidden platform.In experimentation, ambient environmental conditions is kept quite, is stablized.

Experiment is divided into training period and testing period, and 1-4d is training period.During training, rat is put into water back to platform, the escape latency of record rat, namely enters water to the time found required for hidden platform under water from rat, if do not find platform in rat 60s, guides rat to find platform.Escape latency is in 60s, and rat is found or is directed into after on platform and stops 20s, then dries and puts back in cage.Every rat enters water level and puts and convert successively in the direction of the clock, every twice training interval 30s, and the training of the second day comparatively first day quadrant that advances enters water, and the rest may be inferred.5th, 7d is the testing period, removes platform during test.The flat roof area time ratio of rat is all recorded in twice test, and the time of namely swimming across original platform place quadrant in rat 60s accounts for total time the ratio of (60s).

Adopt one factor analysis of variance (One-WayANOVA) analysis, P < 0.05 thinks to have statistical significance.Experimental result is in table 16.

Table 16 is group rat training period average escape latency respectively

Note: ^*p < 0.01, ^#p < 0.05

As shown in Table 16, administration group compares with model group: ^#p < 0.05, after illustrating that rat gives the present composition, its escape latency obviously shortens, and namely the present composition can improve the ability of learning and memory of stress depression rat.

The mensuration of Biochemical Indexes---monoamine oxidase, MAO (MAO), glutathione S-transferase (GST), γ paddy amine acyltransferase (γ-GT)

After Wistar rat animal carries out last process, by rat sacrificed by decapitation, on ice pan, be separated rapidly the cerebral cortex of rat, excision low brain stem.10% brain homogenate is made in normal saline 1: 9 homogenate in ice-water bath with 0.86% pre-cooling after weighing, and after each specimen subpackage several parts, cryogenic refrigerator is frozen for subsequent use.Take out the homogenate for preparing thaw after 3000r/min, centrifugal 15min obtains homogenate supernatant, according to method shown in test kit description, adopts Chromogenic assay to measure the enzymatic activity of MAO, GST, γ-GT respectively.It the results are shown in Table 17.

Table 17 each rat cerebral even slurry biochemical indicator testing result

Note: ^*p < 0.01, #P < 0.05, ##P < 0.01

As shown in Table 17, administration group compares with model group: ^#p < 0.05, ^##p < 0.01, illustrates that the present composition can significantly improve the activity of monoamine oxidase, MAO (MAO), glutathione S-transferase (GST), γ paddy amine acyltransferase (γ-GT).

Conclusion: the present composition has certain protective role to chronic stress depression rat.

Embodiment 7 present composition is tested bacterial meningitis animal protection Effect study

Bacterial meningitis model is set up

Laboratory animal

Because bacterial meningitis sickness rate in infant is higher, be the serious acute infectious diseases of neonatal period, so bacterial meningitis laboratory animal adopts 11 age in days SD neonate rats.

Pathogenic bacterium: GBSIII strain is purchased from Chinese pharmaceutical biological product test sensitivity institute

Dry strain is in serum broth after 37 DEG C of cultivation 24h, and streak inoculation, in blood plate, after 18-24h, selects single bacterium colony in serum broth, receives bacterium to mid log phase.Adopt OD value and 0.5 standard opacity tube (concentration 1 × 10 that turbidimetry adjustment bacterial concentration is corresponding ⁸cfu/ml) identical, show that bacterial concentration is 10 ⁸cfu/ml.With normal saline, antibacterial is diluted to desired concn 10 ⁴cfu/ml.

Model is set up

1) experimental group SD neonate rat 1.0% pentobarbital sodium (15-25mg/kg) intraperitoneal injection of anesthesia.Hold the vertical inserting needle of 10ul puncture needle 2-4mm recess (cerebellomedullary cistern) after neonate rat two have sharp ears line mid point, have after obviously breaking through sense, pumpback is shown in that limpid cerebrospinal fluid (CSF) flows out, and extracts 10ulCSF out and discards.Experimental group slowly injects GSBIII bacteria suspension 10ul.Whole process completes 1, in 2min, postoperative flat sleeping, puts room temperature 25 DEG C, after recovering, puts back to mouse cage and continued to feed by female Mus.After cultivating certain hour, experimental group CSF is extracted out, through process, be divided into the brain injection liquid being mixed with CSF stock solution, cefotaxime+CSF, present composition 500g/kg+CSF, the several difference group of present composition 250mg/kg+CSF, present composition 125mg/kg+CSF.

2) getting newborn SD rat random packet is: blank group, model group, positive controls, administration group 1,2,3.With 1.0% pentobarbital sodium (15-25mg/kg) intraperitoneal injection of anesthesia.Hold the vertical inserting needle of 10ul puncture needle 2-4mm recess (cerebellomedullary cistern) after neonate rat two have sharp ears line mid point, have after obviously breaking through sense, pumpback is shown in that limpid cerebrospinal fluid (CSF) flows out, extract 10ulCSF out to discard, inject mutually commensurability CSF stock solution, cefotaxime+CSF, present composition 500mg/kg+CSF (administration group 1), present composition 250mg/kg+CSF (administration group 2), present composition 125mg/kg+CSF (administration group 3) respectively to each group.Postoperative flat sleeping, put room temperature 25 DEG C, after recovering, put back to mouse cage continued to feed by female Mus.

Clinicing symptom observation

After inoculated bacteria, namely carry out clinical observation, comprising: with or without lip and limbs skin cyanosis, dyspnea, bradykinesia, hypokinesia and tic etc., and occur the time point of symptom the earliest.After inoculated bacteria, 24h adopts neuroethology 5 points of methods of marking processed of Loeffler to assess neonate rat.5 points: when catching back, energy proper motion, stands up in 5s; 4 points, autonomic movement reduces, and can stand up in 5s; 3 points: > 5s stands up; 2 points: can not stand up; 1 point: can not move.For animal of having a convulsion, carry out tic degree evaluation according to the epilepsy SD rat tic grade scale that Racine formulates.This standard is divided into 6 grades, 0 grade: without twitching; I level: facial clonic spasm; II level: on the basis of I level, has the rhythm and pace of moving things to nod; III grade: on the basis of II level, have forelimb clonic spasm; IV grade: on the basis of III grade, have hind leg station power; V level: IV grade basic on fall.

Experimental result is in table 18,19.

Table 18 is inoculated the clinical manifestation of different liquids neonate rat and is compared

As shown in Table 18, after rat gives the present composition, its each clinical symptoms obviously alleviates.

Table 19 is inoculated different liquids 24h neural behavior scoring and is compared

Note: ^*p < 0.01, ^#p < 0.05, ^##p < 0.01

As shown in Table 19, one factor analysis of variance (One-Way ANOVA) is analyzed, and model group compares with blank group: ^*p < 0.01 illustrates modeling success; Administration group compares with model group: ^#p < 0.05, ^##p < 0.01, illustrates that administration group significantly improves compared with the neurobehavioral of model group, and the visible present composition obviously can improve the neurobehavioral of meningitis rat.

The rat still survived after during inoculation 24h, get 10ul CSF through cisternal puncture again before execution and make WBC (leukocyte calculating) counting, count results is as following table 20:

Table 20WBC result of calculation

Note: ^*p < 0.01, ^#p < 0.05, ^##p < 0.01

As shown in Table 20, one factor analysis of variance (One-WayANOVA) is analyzed, and model group compares with blank group: ^*p < 0.01, illustrates modeling success; Administration group compares with model group: ^#p < 0.05, ^##p < 0.01, illustrates that the present composition can obviously reduce rat white blood cell count, reduces the inflammation of meningitis rat.

Conclusion: the antibacterial action that the present composition has had, has certain improvement result to bacterial meningitis animal symptom.

Claims (9)

1. an active ingredient of Chinese herbs compositions, it is made up of jasminoidin, berberine, Radix Ginseng total saponins, it is characterized in that the weight ratio of Radix Ginseng total saponins, berberine, jasminoidin is 3:2:2-0.1.

2. active ingredient of Chinese herbs compositions according to claim 1, is characterized in that the weight ratio of Radix Ginseng total saponins, berberine, jasminoidin is 3:2:2-0.5.

3. active ingredient of Chinese herbs compositions according to claim 2, is characterized in that the weight ratio of Radix Ginseng total saponins, berberine, jasminoidin is 3:2:2.

4. a pharmaceutical preparation, it comprises compositions according to claim 1 as active component and any one or multiple pharmaceutically acceptable carrier.

5. pharmaceutical preparation according to claim 4, it can be made into the pharmaceutically acceptable dosage form of any one.

6. the purposes of compositions according to claim 1 in the medicine of preparation treatment or prevention brain and nerve injury, nervus retrogression encephalopathy, bacterial meningitis or depression.

7. purposes according to claim 6, wherein said brain and nerve injury are cerebrovascular dementia or ischemic cerebrovascular.

8. purposes according to claim 7, wherein said ischemic cerebrovascular is the outbreak of transience local cerebral ischemia, cerebral infarction or apoplexy.

9. purposes according to claim 6, wherein said nervus retrogression encephalopathy is senile dementia or Parkinson's disease.