WO2013004177A1

WO2013004177A1 - Composition of active ingredient of traditional chinese medicine and use thereof

Info

Publication number: WO2013004177A1
Application number: PCT/CN2012/078139
Authority: WO
Inventors: 杨洪军; 李韶菁; 王永炎; 黄璐琦; 付梅红
Original assignee: 中国中医科学院中药研究所
Priority date: 2011-07-05
Filing date: 2012-07-03
Publication date: 2013-01-10
Also published as: CN102861117A; CN103108639B; CN103108639A

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

Traditional Chinese medicine active ingredient composition and use thereof

Technical field

The present invention relates to a traditional Chinese medicine active ingredient composition comprising geniposide or geniposide, berberine, ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition. The invention further relates to the use of the composition for the manufacture of a medicament for the treatment or prevention of brain and nerve damage, neurodegenerative encephalopathy, bacterial or viral encephalitis, or depression. Background technique

Traditional Chinese medicine theory attaches importance to the overall concept and syndrome differentiation. Traditional Chinese medicine theory believes that qi deficiency and blood stasis is an important pathogenesis of stroke. Qi deficiency is the basis of this disease. Blood stasis is the basic pathogenesis of this disease. It is the core of the disease's development and development. The basic method of treating this disease. Qi deficiency and blood stasis are only seen in the traditional sequelae of sequelae. Yiqihuoxue method is only used for sequelae patients. Qing Dynasty famous doctor Wang Qingren Bu Yang Huan Wu Tang is the representative prescription (Zhang Meikui, Yin Ling, Zhang Xuewen Bu Yang Huan Wu Tang) Research progress in the treatment of ischemic stroke, Chinese Journal of Basic Medicine in Traditional Chinese Medicine, 2002, 8 (9): 74-78). The use of Yiqi, detoxification and related Chinese medicine to treat brain and nerve damage diseases has not been reported.

Senile dementia (SD), also known as Alzheimer disease (Alzheimer disease,

AD) is a central nervous system degenerative disease characterized by progressive cognitive impairment and memory impairment, with varying degrees of abnormalities in sports, cognition, language, and personality. The clinical manifestation of AD is dementia syndrome, characterized by memory loss and personality degeneration. The main pathological changes in AD are cerebral cortical atrophy, loss of nerve cells and synapses, neurofibrillary tangles, extracellular senile plaques, and cerebral vascular amyloidosis. There is currently no fundamental treatment for Alzheimer's disease. The current molecular mechanisms and related botanicals for the treatment of senile dementia are: 1) acetylcholinesterase inhibitor scutellarin; 2) excitatory amino acid and peptide transmitter regulators, such as auxin; 3) interference with beta-starch Protein (Αβ) fiber formation and deposition, such as ginkgo flavonoids, salvianolic acid, emodin, timosaponin and so on. 4) Antioxidant and free radical scavenging drugs, Salvia miltiorrhiza water extract salvianolic acid, tea extract tea polyphenols, Ginkgo biloba extract, ginsenoside, ginkgolides, etc.; 5) Improve microcirculation, such as tanshinone and danshensu ; 6) Calcium antagonists, such as apigenin.

Vascular Dementia (VD) is a general term for dementia syndrome caused by brain damage caused by a series of cerebrovascular factors. It is mainly caused by memory and cognitive dysfunction on the basis of cerebrovascular disease, or A disease associated with acquired intelligence, continuous impairment of language, spatial skills, and emotional or personality disorders. Clinically, it is characterized by memory loss, calculation, judgment, orientation and other progressive mental and behavioral disorders. It is a common disease and frequently-occurring disease in the middle-aged and elderly population. The exact pathological mechanism of VD is not fully understood. It is generally believed that cerebrovascular disease causes local brain tissue inflammation, metabolic disorders and oxidative stress loss, damages brain tissue of sufficient capacity, and seriously impairs advanced neurological functions such as memory and cognition. Especially when the lesion involves the frontal lobe, temporal lobe and limbic system, the symptoms of dementia are more likely to occur. The traditional Chinese medicine compound for treating VD is: Huanglian Jiedu Decoction, Danggui Shaoyao San, Dihuang Yinzi, Tianzhi Granule, Congsheng Capsule, Shenmao Yizhi Capsule, Liuwei Dihuang Decoction, Jiawei Guipi Decoction, Buyang Huanwu Decoction, etc. The use of Yiqi, detoxification and related Chinese medicine has not been reported.

Parkinson disease (PD), also known as tremor paralysis, is a common central nervous system disease in middle-aged and elderly people. The clinical manifestations include static tremor, bradykinesia, myotonia and posture gait abnormalities. The pathogenesis is a dopaminergic (DA) neurological disorder in the central nigrostriatal pathway, leading to a loss of striatum DA, causing somatic motor dysfunction. When dopamine is reduced, acetylcholine is overexcited, and the two transmitters lose balance, causing nerves and muscle tissue to involuntarily produce symptoms such as rigidity and tremor. At present, there is no cure for Parkinson's disease. The western medicine levodopa (Medopa), bromocriptine, and ergocarpine are commonly used drugs in the clinic, but the effect is unstable, the side effects are large, and the long-term efficacy is not certain. Commonly used traditional Chinese medicine treatment methods and commonly used drugs are as follows: 1) Bugan deficiency, Zhengan Xifeng: Tianma, antler gum, asparagus, Ophiopogon japonicus, armor, turtle shell, raw white peony, licorice, etc.; 2) kidney deficiency Filling the qi and qi: 萸肉, Schisandra; 3) Complementing the blood, but also replenishing the brain: using Dayun, Angelica, longan meat, jujube, etc.; 4) Promoting blood circulation: Sanqi, pangolin, salvia, scorpion . Chen Jianzong et al. found that Chinese herbal medicines consisting of medlar, Cistanche, and Polygonum multiflorum can improve the content of DA, 3, 4-dihydroxyphenylacetic acid (DOPAC), NE, 5-HT in the striatum of PD rats. It has a certain protective effect on the nucleus of the substantia nigra, blue spot, and dorsal nucleus (Chen Jianzong, Guo Jianying, Sun Jing, etc., 40 cases of vascular Parkinson's syndrome treated by Peibu Ganshen Qufeng Tongluo Method, Journal of Traditional Chinese Medicine , 2002, 43(7): 528-529). He Jiancheng et al. found that Chinese herbal medicines with yin and tonifying kidney, collaterals and detoxification, which are composed of medlar, mulberry parasitic, tianma, uncaria, and silkworm, can significantly reduce the release of reactive oxygen species (ROS) and propylene from the brain of PD rats. The content of aldehyde (MDA) increases the content of free radical scavenging systems such as glutathione (GSH), glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD), which improves the body's antioxidant capacity. To alleviate the effects of free radicals on the body damage (He Jian, Yuan Canxing, Li Yaming, etc., the effect of nourishing liver, kidney, collaterals and detoxification Chinese medicine on oxidative stress in rat model of Parkinson's disease, Chinese Journal of New Drugs and Clinical Medicine, 2003, 22(3) ): 160-163). Xu Li and other observations of Xifeng Zhiyu Capsule (Danggui, Baiqi, Qi, Quanqi, etc., which are composed of Ziyin Yangxue and Xifeng Tongluo Traditional Chinese Medicine) for the treatment of experimental hypoxic experimental PD mice, which found that it can significantly increase PD The content of DA, DOPAC and high vanillic acid (HVA) in the brain of rats can reduce the intensity of limb tremor in mice and shorten the duration of tremor. Therefore, it is believed that Xifeng Zhiyu Capsule can prevent and treat PD (Xu Li, Wei Cuiyi, Liu Jianxun, etc. The preventive and therapeutic effects of Xifeng Zhizhi Capsule on experimental Parkinson's disease in mice, New Chinese Medicine and Clinical Pharmacology, 2005, 2: 87-90) The use of Yiqi, detoxification and related Chinese medicine has not been reported.

Depression usually refers to a kind of emotional disorder, mainly characterized by low mood, slow thinking, and reduced language action. It is mainly characterized by depression, decreased interest, pessimism, slow thinking, lack of initiative, and self-blame. Self-sin, diet, poor sleep, worry about having various diseases, feeling multiple discomforts in the body, serious suicidal thoughts and behaviors. Depression has the characteristics of high incidence, high prevalence, high recurrence rate, high suicide rate, low awareness rate and low treatment rate. The etiology of depression is complex. The pathological mechanisms hypothesis that has been studied are as follows: 1) Metabolic abnormalities of monoamine neurotransmitters, which are caused by insufficient monoamine transmitters in certain parts of the brain; 2) Changes in body function, antidepressant inhibition of presynaptic monoamine transmitter intake is an immediate effect, while post-synaptic receptor sensitivity is a chronic adaptive change; 3) nerve internal division Disorders, studies have found that depression occurs and hypothalamic-pituitary-adrenal axis (HPA), hypothalamic-pituitary-thyroid axis (HPT), hypothalamic-pituitary-growth hormone axis (HPGH) and other dysfunction and serum cholesterol levels Lowering, serum estrogen levels are associated with decreased levels. At present, antidepressants mainly have three characteristics to varying degrees, namely, they have an stimulating effect on mental exercise, enhance internal drive, active emotions and anti-anxiety effects. It is mainly divided into five major categories: monoamine oxidase inhibitors, tricyclic antidepressants (TCAs, such as Datlan), tetracyclic antidepressants, new antidepressants such as selective 5-tryptamine reuptake inhibitors ( SSRIs, such as fluoxetine), selective norepinephrine reuptake inhibitors, serotonin and norepinephrine reuptake dual inhibitors (SNRIs such as venlafaxine), norepinephrine and specific 5- A tryptamine reuptake inhibitor (NaSSAs, such as mirtazapine), norepinephrine and dopamine reuptake inhibitors, botanical extracts, and the like. However, most of them have the characteristics of narrow anti-depression spectrum, large side effects, high drug price and easy recurrence.

Meningitis can be divided into two categories: bacterial and sterilized. Bacterial meningitis is inflammation caused by bacterial infection of the pia mater. It is a serious intracranial infectious disease, often accompanied by encephalitis and brain abscess. From the perspective of pathogens, there are two major categories, one is meningitis of various purulent bacterial infections (abbreviation: brain), such as meningitis caused by meningococcal, pneumococcal, influenza, Haemophilus, etc. The other type is meningitis infected with Mycobacterium tuberculosis. Clinical medication is mainly antibiotics combined with hormone therapy. Aseptic meningitis refers to cerebrospinal fluid smears and bacterial cultures that are recessive meningitis. Generally, aseptic meningitis is caused by viral infection. Viral encephalitis refers to inflammation of the brain parenchyma caused by viral infections, often manifested as fever, headache, convulsions, disturbance of consciousness, and meningeal irritation, which can cause focal damage to the central nervous system. Viral encephalitis has a poor prognosis, high mortality, and often has serious sequelae. Commonly used drugs are 1) hormones, such as the adrenal cortex hormone dexamethasone; 2) antiviral steroids, such as acyclovir or adenosine; 3) antiviral cytokines, interferon; 4) nutrition Such as citicoline, vitamin B6, vitamin E, brain rehabilitation, pantothenic acid, etc. to improve brain metabolism, patients with seizures should also be combined with anti-epileptic drugs, patients with mental exercise excitement can use mentally stable drugs, etc. .

There are still defects in the treatment of brain or nerve damage diseases, such as side effects, poor efficacy, and inability to cure. Because brain disease is a complex pathophysiological process involving multiple links, multiple factors, and multiple pathways, traditional Chinese medicine compound preparations can have multiple neuroprotective effects, with multi-target, multi-directional regulation, and clinical significance of prevention and treatment. It should be expected to be a more effective brain and neuroprotective drug.

Various ginseng is a good medicine for tonic and is often used in Chinese medicine. "Shen Nong's Herbal Classic" records ginseng. "The main remedy for the five internal organs, the spirit of the spirit, the soul of the soul, the horror, the evil spirits, ..., "Happy Puzzle" is considered to be a kind of Chinese herbal medicine with various pharmacological effects. Its main active ingredients for brain and nerve damage have been reported in many cases, such as prevention and treatment of Alzheimer's disease, Parkinson's disease, depression, and cerebral infarction. Other diseases caused by cerebral ischemia and cerebral ischemia (Jiangshan, Jiang Zhenglin, Zeng Yinming, Research on the effects of nine kinds of ginsenosides against cerebral ischemic injury, Chinese Pharmacology and Clinical Medicine 2007; 23(2): 19-21; Xu Wei, Liu Chunqing, Ma Tao et al. Protective effects of ginsenoside Re on dopaminergic neurons in MPTP-induced Parkinson's disease model mice, Journal of Shenyang Pharmaceutical University, 2005, 22

(1): 36-44; ginsenoside composition and preparation method, invention patent, application number 200610046083.3; compound 20S-ginsenoside R 2 in preparation of antidepressant medicine, invention patent, application number 200810043537.0). It has the effects of diarrhea, diarrhea, dampness, detoxification, and insecticidal effects. Modern pharmacological studies have shown that in addition to its strong antibacterial activity, it also has anti-inflammatory, hypolipidemic, hypoglycemic, anti-tumor, anti-ulcer, A variety of pharmacological effects such as neuroprotection. It has been reported that Coptis has protective effects in cerebral ischemia and reperfusion injury through anti-inflammatory effects (Ki-Yeon Yool, In Koo Hwang2, Jong Dai Kim3, etc., Antiinflammatory Effect of the Ethanol Extract of Berberis koreana in a Gerbil Model of Cerebral Ischemia /Reperfusion Phytother. Res. 2008, 22, 1527-1532 ).

The scorpion is the fruit of the Gardenia j匪 inoides Ellis. It is bitter and cold and non-toxic. Into the heart, liver, lungs, stomach. It has the effects of purging fire, removing heat, diuresis, cooling blood and detoxification. Indications of fever, irritability, jaundice, gonorrhea, diabetes, red eyes, sore throat, vomiting blood, blood stasis, blood stasis, hematuria, heat sores, sprains and swelling. Clinically used for acute jaundice hepatitis, hemostasis, contusion and other diseases. Modern pharmacological studies have shown that in addition to traditional pharmacological effects such as antipyretic, antibacterial, sedative, analgesic, choleretic, anti-inflammatory, hemostasis, liver protection, and soft tissue damage repair, medlar has anti-oxidation, anti-asthma, and hypolipidemic effects. , antihypertensive, anti-allergic, anti-atherosclerosis, anti-viral, anti-depressive anxiety, nervous system protection, etc. (Study on the chemical constituents and pharmacological effects of gardenia, North Pharmaceutical, 2008, 5 (3): 44-45, 51; Huang Shisun, Wu Yuyue, Overview of Modern Pharmacological Research and Clinical Application of Xunzi, Internal Medicine, 2010, 5 (5): 534-536; Liu Guomin, Guo Suhua, Cheng Weiming, New Progress in the Pharmacological Action and Mechanism of Gardenia, Strait Pharmaceuticals, 2008, 20 (1 1 ): 8-10; Use of Gardenia Extract in the Preparation of Drugs for Prevention and/or Treatment of Depression Worries, Chinese Invention Patent, Application No. 200810016679.8).

The classic compound Huanglian Jiedu Decoction shared by Huanglian and Scorpion is originally contained in the "Outside Taiwan Secrets". It is composed of berberine, Phellodendron, Astragalus and Scorpion. It is regarded as the representative of detoxification and detoxification. It has the effect of purging fire and detoxification. Practical heat and poison, Sanjiao hot Sheng card. It has been reported that Huanglian Jiedu Decoction can reduce brain damage and inflammatory infiltration in cerebral ischemia and reperfusion (Young Sun Hwang a, Chan Young Shin b, Youngbuhm Huh et al., Hwangryun-Hae-Dok-tang (Huanglian). -Jie-Du-Tang) extract and its constituents reduce ischemia-reperfusion brain injury and neutrophil in filtration in rats. Life Sciences 2002, 71 : 2105-21 17).

The inventors have noted that there is still a lack of pharmaceutical preparations that can be used for the treatment of brain and nerve damage diseases. Summary of the invention

The composition of the invention is derived from an effective traditional Chinese medicine compound Yiqi Jiedu Decoction for clinical treatment of stroke. Yiqi Jiedu Recipe is a key pathogenesis for "poisoning and cerebral palsy". It is proposed by using syndrome differentiation and treatment to correct Chinese medicine encephalopathy. Yiqi Jiedu Recipe is a new traditional Chinese medicine compound preparation that has not been reported before. Its active ingredients have not been reported for the prevention and treatment of brain and nerve damage diseases in Chinese medicine. Yiqi Jiedu Recipe consists of ginseng, berberine and medlar. Among them, ginseng is a medicinal herb, it is a great qi, and it can replenish the five viscera. It is very suitable for the treatment of qi deficiency; berberine is a drug, it is a good product of diarrhea and detoxification, compatible with ginseng, one supplement and one diarrhea, The scorpion is the adjuvant medicine, the medicine is good for clearing the heart, removing blood, detoxifying, assisting the diarrhea and detoxification of berberine, When the sexual flexion goes down, the evil spirits are driven out of the urine, which makes the evil of heat and poison have a way out. This three-flavor compatibility can effectively detoxify evil on the basis of righting up, and plays a very good synergistic therapeutic effect.

Excessive release of excitatory amino acids and neurotransmitter abnormalities, oxidative stress, free radicals, inflammatory cascades, mitochondrial dysfunction, etc. are common pathological mechanisms of brain injury, ginseng grows in qi, neuroprotection, berberine and scorpion are longer than detoxification , the three compatibility can be rationally compatible and mutually complementary.

Accordingly, the technical problem to be solved by the present invention is to provide a composition of a traditional Chinese medicine active ingredient capable of preventing or treating a brain and nerve damage disease.

The present invention relates to a traditional Chinese medicine active ingredient composition comprising geniposide or geniposide, berberine, ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition.

The composition according to the present invention as described above, wherein the active ingredient of the traditional Chinese medicine is calculated in accordance with the total weight of 100 parts including the following parts by weight: ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition, 1- 99 parts, berberine, 1-99 parts, geniposide or geniposide, 1-99 parts; preferably ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition, 2-75 parts, berberine, 2-75 parts, geniposide or geniposide, 2-75 parts; more preferably ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition, 3-50 parts, berberine, 3-50 parts, Geniposide or geniposide, 3-50 parts.

The composition according to the present invention as described above, wherein the weight ratio of ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition, berberine, geniposide or geniposide is 3: 2: 2-0.1 Preferably 3:2: 2-1, more preferably 3:2:2.

The composition according to the present invention as described above comprises a traditional Chinese medicine active ingredient and any one or more pharmaceutically acceptable carriers such as an antioxidant, a complexing agent, a filler, a matrix material and the like.

The composition according to the present invention as described above may be formulated into any pharmaceutically acceptable dosage form, such as a tablet, a granule, a capsule, an injection such as a powder, according to a specific need and a pharmaceutically acceptable carrier. Freeze dry powder, infusion, etc.

The invention further relates to a composition as described above for the treatment or prevention of brain and nerve damage, such as cerebral vascular dementia or ischemic cerebrovascular disease (eg transient local ischemic attack, cerebral infarction or stroke), neurodegenerative Use in diseases of the brain, such as Alzheimer's disease or Parkinson's disease, bacterial or viral encephalitis, or depression. DRAWINGS

Figure 1 : TTC staining results of cerebral ischemic area of cerebral ischemia rats after 24 hours of MCAO surgery, A: sham operation group; B: model group; C: positive control group (ginkgo biloba extract Egb761 tablets) Group) (4 mg/kg); DF: administration group 3, 2, l (62.5 mg/kg).

Figure 2: Effect of the composition of the invention in Example 1 on the volume of cerebral infarction in rats 12 h and 24 h after MCAO surgery (n = 10). detailed description

The invention will be further illustrated by the following examples in which the advantages and features of the invention will become more apparent. However, these examples are merely exemplary and do not constitute any limitation on the scope of the invention. It will be understood by those skilled in the art that the details and forms of the present invention may be modified or substituted without departing from the spirit and scope of the invention, and such modifications and substitutions fall within the scope of the invention.

Preparation example of active ingredients of traditional Chinese medicine

Berberine extraction process:

Take the Rhizoma Coptidis, smash it, place it in the Soxhlet extractor, add 95% ethanol and heat to reflux until the extract is colorless. Concentrate 95% ethanol extract to a paste, add 1% acetic acid, dissolve by heating, remove by suction. Insoluble matter, concentrated hydrochloric acid was added dropwise to the solution until the solution became turbid, and left to cool, that is, yellow acicular berberine hydrochloride was precipitated, suction filtered, the crystal was washed twice with ice water, and then washed once with acetone to accelerate Dry to give a crude product. The crude product is dissolved in water, adjusted to a pH of 8.5 to 9.8 with lime milk, cooled, and the impurities are filtered off, and cooling is continued until the room temperature is below, and crystals are precipitated, suction filtered, and purified.

Extraction process of total saponins from ginseng stems and leaves:

Take ginseng stems and leaves, cut into l~2cm sections, add water to cook 2 times, the first 2 hours, the second 1.5 hours, combine the filtrate, pass D101 type macroporous resin, water elute to colorless, then use 60 After elution with % ethanol, a 60% ethanol eluate was collected, and the eluate was concentrated to a clear paste having a relative density of 1.06 to 1.08 (80 ° C), and dried and pulverized.

Extraction process of geniposide or geniposide:

Take the scorpion medicinal material, smash it, add water to cook for 3 times, 1.5 hours each time, filter the decoction, and combine the filtrate. Pass the AB-8 macroporous resin, elute with water, 5% ethanol to colorless, then use 10 The ethanol solution was eluted, and the 10% ethanol eluate was collected. The eluate was concentrated to a clear paste having a relative density of 1.06 to 1.08 (80 ° C), and dried and pulverized.

The ginsenoside monomer of the present invention may be ginsenoside Rgl, Rbl, Re, Rfl, Rd, R (R l and R 2).

The ginsenoside monomers of this patent include, but are not limited to, obtained by the following methods:

(I) Preparation of ginsenoside monomer Rfl is prepared according to the patent method (medical use of ginsenoside F1, application number 200610008477.X);

(II) Preparation of ginsenoside monomer Re is prepared according to the patent method (new method and preparation method of ginsenoside Re medicine, application number 2004110021133.6);

(III) Preparation of ginsenoside monomer Rgl is prepared according to the patent method (the use of ginsenoside Rgl, application number 200810227643.4);

(IV) Preparation of ginsenoside monomer Rbl is prepared according to the patent method (a preparation method of ginsenoside Rbl, application No. 200710178212.9); (V) Preparation of ginsenoside monomer Rh (R l and Rh2) is prepared according to the patent method (ginsenoside R extract and preparation method, application number 200810043537.0);

(VI) Preparation of ginsenoside monomer Rd by reference literature method (Liu Deyu, Cao Yu. Method for rapid separation and extraction of ginsenoside Rd from Sanqi medicinal materials, Chinese medicinal materials, 2006, 29 (3 ): 247-249 ).

The above ginseng total saponins and ginseng monomers are known as pharmaceutically active substances, and can be used as a raw material for pharmaceutical preparations by mixing ginseng total saponins or the above various ginsenoside monomers or two or more kinds of ginsenoside monomers. After being mixed with berberine and geniposide in a certain proportion as follows, according to the conventional preparation method, it is prepared into any clinically applicable agent, such as an oral solid preparation such as a tablet, a capsule, a granule, a dropping pill, orally. Liquid preparation, injection, and the like. In the preparation of the above preparation, the mixture prepared above may be used in combination with any suitable excipient for the preparation of a clinical preparation to prepare a pharmaceutical preparation.

Pharmaceutical composition ratio method

(1) Calculated according to the ratio of parts by weight, 30 parts of total ginsenoside prepared according to the above method, 20 parts of berberine and 5-20 parts of geniposide are mixed as raw materials for pharmaceutical preparations;

(2) Calculated according to the ratio of parts by weight, 30 parts of ginsenoside monomer, 20 parts of berberine and 5-20 parts of geniposide prepared according to the above method are used as raw materials for pharmaceutical preparations;

(3) Calculated according to the weight ratio, 30 parts of two or more ginsenoside monomer mixtures prepared according to the above method, 20 parts of berberine and 5-20 parts of geniposide are mixed as a pharmaceutical preparation. Raw material medicine.

The pharmaceutical composition of the present invention comprises a composition in combination with berberine and geniposide and a pharmaceutically acceptable excipient or carrier according to the above (1) - (3) ratio method.

The compound of any one of the above (I) - (VI) of the present invention is added to various excipients or pharmaceutically acceptable excipients or carriers required for preparation of different dosage forms, and the conventional pharmaceutical preparation method is used. It can be prepared into any suitable clinical preparation, and can be, for example, an injection (powder needle, lyophilized powder, water needle, infusion, etc.), a tablet, an oral solution, a granule, a capsule, a soft capsule, a dropping pill, and the like. Wherein, the auxiliary material may be an antioxidant complexing agent, a filler, a matrix material, etc.; the pharmaceutically acceptable carrier is xylitol, mannitol, lactose, fructose, dextran, glucose, polyethylene One or more of pyrrolidone, low molecular weight dextran, sodium chloride, calcium gluconate or calcium phosphate, preferably mannitol or lactose. Example 1 Experimental study on the protective effect of the composition of the present invention on acute focal cerebral ischemia

Establishment of an animal model of acute focal cerebral ischemia (middle cerebral artery occlusion (MCAO) surgery) Male SD rats weighing 250-270 g, refer to Zea Longa et al (Longa EZ, Weinstein PR, Carlson S, Cummins R. Reversible Middle cerebral artery occlusion without craniectomy in rats. Stroke 1989; 20: 84-91) established a method for blocking the occlusion of the middle cerebral artery in rats. 10% chloral hydrate solution 400 mg/kg, anesthetized animals were injected intraperitoneally. The rat was fixed in the supine position, the skin was cut in the middle of the neck, and the layers of tissue were bluntly separated to expose the right common carotid artery (CCA). Separate to the internal carotid artery (ICA), external carotid artery (ECA) after a bifurcation, carefully separate to avoid damage to the vagus nerve and trachea, set the line for use. Isolation of the ipsilateral external carotid artery, about 0.8cm in ECA Ligation. An arterial clip was placed at the proximal end of the CCA. A "V"-shaped incision with a diameter of about 2 mm was made between the ECA ligation and the bifurcation. The nylon thread was gently inserted into the CCA from the incision, and the artery clip was released. The inner and outer carotid bifurcations enter the internal carotid artery, and the nylon thread is slowly pushed into the cranial direction of the ICA. The insertion depth is about 18.5±0.5 mm to the micro-inductance, so that the nylon thread end passes through the MCA start point. The thinner anterior cerebral artery, at this time, achieves blood flow obstruction of the left middle cerebral artery, ligation of ICA to fix the nylon thread and prevent bleeding, suture layer by layer, and the nylon thread stump remains l cm longer than the skin.

The sham operation group only performed preoperative anesthesia and vascular separation, without ligation and introduction of a wire plug. The room temperature was maintained at 24-25 °C during the operation, and the biological function test system was used for animal breathing and ECG monitoring.

Animal grouping and administration

SD rats were randomly divided into six groups: sham operation group, model group, Ginkgo biloba extract tablets positive control group (Ginado Ginkgo biloba extract tablets, Egb761, Dr. Weimar Shupei, Germany), the combination of the present invention Drug administration group 1, 2, 3. Positive control group administration method: Take 1 piece of Ginkgo biloba extract tablets (40mg) dissolved in 40ml normal saline, prepare lmg/mL, and give SD rats 1mL by gavage, the dosage is 4 mg/kg. Administration method of the drug-administered group: Take ginseng total saponin: berberine: geniposide according to the weight ratio of 3:2:2 (administration group 1), 3:2:0.1 (administration group 2), 3:2: 0.5 (administration group 3), 31.25 mg/mL was prepared with physiological saline, and 0.5 mL was administered by intragastric administration at a dose of 62.5 mg/kg. The drug-administered group and the positive control group were intragastrically administered 10 minutes before the anesthesia, and the animals were sacrificed at 4, 12, and 24 hours after surgery, and the following indicators were tested.

Neurobehavioral Bederson's functional evaluation

Neurobehavioral observations were performed prior to sacrifice, according to the method of Bederson et al. (Bederson JB, Pitts LH, Tsuji M, et al., Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. Stroke 1986 17:472-476), take the rat tail off the ground about 1 foot, observe the condition of the two forelimbs; place the rats on the level ground, push the shoulders, observe the difference of the resistance on both sides; the rats are placed on the ground, observe Its walking situation. Using a five-point scale (0-4 points), the higher the score, the more serious the neurobehavioral damage. The statistical results of Bederson's functional scores are shown in Table 1. The data of each group were compared using the rank sum test, and Ρ<0.05 was considered statistically significant.

Table IBederson's functional score statistics (±sd, n=10) sham operation model positive pair

Item Level Drug Delivery Group 1 Drug Delivery Group 2 Drug Delivery Group 3 Groups

0 6

1 1 4

2 1 3 2

12h-l 3 5 2

4

Rank

P value # #

0 7 4 4 5

1 2 0 4 2 1

2 1 2 2

12h-2 3 4 1 1 1

4

Rank

P value ## # ## ##

0 10 0 5 5 1 0

1 0 4 4 3 6 7

2 0 0 0 1 3 2

24h-l 3 0 1 0 0 0 0

4 0 3 0 0 0 0

Rank

P value ## ## ## #

0 8 0 3 5 5 3

1 2 4 3 2 2

2 4 0 0 0 1

24h-2 3 0 0 0 0 0

4 2 0 0 0 0

Rank

P value # ## ## ##心

Product P value ## ## ## 分

Note: Non-parametric rank sum test, test, compared with control group: : "P<0.01; compared with model group: #P

<0.05, ##P<0.01

Conclusion: It can be seen from Table 1 that the model group is statistically different from the sham operation group, indicating that the modeling is successful (Ρ<0.01). The positive control group (3 mg/kg) and the drug-administered group 1, 2, 3 significantly improved the neurological symptoms at 12h and 24h after focal cerebral ischemia in animals (P<0.05, P<0.01).

Evaluation of the improvement of cerebral infarction volume after cerebral ischemia

After functional scoring, the rats were decapitated and the brain tissue was quickly placed in a refrigerator at -20 °C. After lOmin, the room temperature was set. The olfactory bulb, cerebellum and low brain stem were removed and cut into 2 mm crowns according to the brain localization map. Six brains were continuously coronal sections. Then quickly place the brain slices in 5ml containing 2% TTC (triphenyltetrazolium chloride) and

In a 0.1 ml solution, incubate at 37 ° C for 30 min in the dark, and flip the brain slices once every 5 min. After TTC staining, the normal tissue was rose red, and the infarcted tissue was unstained and white. Each group of brain slices was arranged neatly, photographed and saved, and processed by image analysis system software and counted. The infarct area of each brain slice was calculated, multiplied by the thickness of each brain slice by 2 mm, and the infarct area of each brain slice of each animal was multiplied by The thickness is added, which is the volume of cerebral infarction.

Conclusion: As shown in Figures 1 and 2, after cerebral ischemic injury in rats, no cerebral infarction occurred in the sham operation group, and the infarct area appeared in the model group and the TTC stained grayish white. The administration group 1, 2, 3 can significantly reduce the gray area of the cerebral infarction, reduce the volume of cerebral infarction, and significantly improve the cerebral infarction. Evaluation of the improvement of regional cerebral blood flow after cerebral ischemia in rats with middle cerebral artery occlusion (MCAO) The MCAO procedure was as described above, and the drug was given 15 min before anesthesia with chloral hydrate.

Animal grouping and administration method

SD rats were randomly divided into the following groups: sham operation group, model group, Ginkgo biloba extract tablets positive control group (Ginado Ginkgo biloba extract tablets, Egb761, Dr. Weimar Shupei, Germany), the present invention Composition administration group (ginseng total saponin: berberine: geniposide in a weight ratio of 3:2:2). Positive control group administration method: Take apricot leaf extract tablets (40mg) dissolved in 40ml normal saline, formulated into lmg/mL, and intragastrically administered to SD rats lmL, the dose is 4 mg / kg. Administration method of the drug-administered group: The drug-administered group was prepared into 0.25 mg/mL, 1.25 mg/mL, and B 6.25 mg/mL, respectively, according to the compatibility ratio, and the intragastric administration was 1 mL. The three doses were from high to low. l mg/kg, 5 mg/kg and 25 mg/kg. The animal was placed supine on the operating table, and before the anesthesia was performed without MCAO surgery, the blood flow value was recorded within 15 min using the laser speckle blood flow monitoring video system (PariCam PSI), and the average value was used as cerebral blood. The baseline value of the stream. After completing the MCAO operation, the line was inserted into the cranium. When the blood flow value suddenly dropped to 40%-60% of the baseline value, the blood flow of the middle cerebral artery was blocked, and each group was administered with the model group. The blood flow value before surgery was the basic value of the group (100%), and the blood flow value after surgery was expressed as a percentage of the base value. The LDF value was measured at the same position 30 min after the operation, and the SD rats were monitored for blood flow changes before and after MCAO operation. This system can not only grasp the spatial distribution of blood perfusion, but also clearly see the real-time dynamic changes of blood flow. The results of the composition of the present invention on cerebral blood flow after SDAO operation in SD rats are shown in Table 2.

The mean blood perfusion (PU) is calculated as:

Mean blood flow volume (PU) = blood cell movement velocity (V) X blood cell concentration (C)

The regional cerebral blood flow rate of change was calculated using the following formula: Percent change = ^TQItteated _ ^TQI∞ntto1 x 100%

TOIcontrol

Among them, TOI _∞ntol is the average blood flow volume of the auricle before administration (before surgery), and TOIt^w is the post-administration ear composition. The composition of the present invention exerts cerebral blood flow after SDAO operation in SD rats (n=5) )

Group agent L (mg-kg ^-1 ) mean blood flow reduction rate (%) sham operation group - 0

Model group - 51.95±4.71

Positive control group 4 22.09±7.8**

Drug administration group 1 32.96±5.17

5 21.71±2.16

25 14.93±3.13

Note: Non-parametric rank sum test, test, compared with control group: "P <0.01; compared with model group: ^## P < 0.01

Conclusion: By monitoring the change of regional cerebral blood perfusion (rCBF) within half an hour after ischemia, the positive drug group (4 mg/kg) and the high, medium and low doses of the drug-administered group were administered within half an hour after MCAO surgery. L mg/kg, 5 mg/kg, 25 mg/kg) can significantly improve the reduced regional cerebral blood flow after focal cerebral ischemia, and the low-dose group of the composition of the present invention (l mg/kg, 32.96±5.17) ") still showed better improvement in local blood perfusion, compared with the model group, statistically significant (P <0.01), and showed a good concentration dependence. This shows that the composition of the present invention can quickly pass through The blood-brain barrier may exert better vascular protection by dilating blood vessels and increasing local blood perfusion.

From the above experiments, it is known that the composition of the present invention has a good protective effect against acute cerebral ischemic injury. Example 2 Preparation of the composition of the present invention for ischemia-reperfusion injury in animals with focal cerebral ischemia Preparation of middle cerebral artery occlusion (MCAO) model of ischemia-reperfusion

Male Sprague-Dawley rats weighing 250-270 g were used with reference to Zea Longa et al. (Longa EZ, Weinstein PR, Carlson S, Cummins R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 1989; 20: 84-91). The method of blocking the wire in the middle cerebral artery of the rat is appropriately improved.

10% chloral hydrate solution 400 mg/kg, anesthetized animals were injected intraperitoneally. The rat was fixed in the supine position, the skin was cut in the middle of the neck, and the layers of tissue were bluntly separated to expose the right common carotid artery (CCA;). Separate to the internal carotid artery (ICA), the external carotid artery (ECA) after the bifurcation, carefully separate to avoid damage to the vagus nerve and trachea, set the line for later use. The ipsilateral external carotid artery was isolated and ligated at approximately 0.8 cm from the ECA. An arterial clip was placed at the proximal end of the CCA. A "V"-shaped incision with a diameter of about 2 mm was made between the ECA ligation and the bifurcation. The nylon thread was gently inserted into the CCA from the incision, and the artery clip was released. The inner and outer carotid bifurcations enter the internal carotid artery, and the nylon thread is slowly pushed into the cranial direction of the ICA. The insertion depth is about 18.5±0.5 mm to the micro-inductance, so that the nylon thread end passes through the MCA start point. a thinner anterior cerebral artery, At this point, the blood flow obstruction of the left middle cerebral artery is achieved, the ICA is ligated to fix the nylon thread and the bleeding is prevented, and the suture is layer by layer, and the nylon thread stump is longer than the skin.

Animal grouping and administration

SD rats were randomly divided into the following groups: sham operation group, model group, Ginkgo biloba extract tablets positive control group (Ginado Ginkgo biloba extract tablets, Egb761, Dr. Weimar Shupei, Germany) (4mg/ Kg), the composition of the composition of the present invention (ginseng total saponin: berberine: geniposide in a weight ratio of 3:2:2). Positive control group administration method: Take 1 piece of Ginkgo biloba extract tablets (40mg) dissolved in 40ml normal saline, prepare 1mg/mL, and give SD rats 1mL intragastrically 3h after operation, the dosage is 4mg/ Kg. Dosing method of the drug-administered group: The drug-administered group was prepared into 0.25 mg/mL, 1.25 mg/mL, and 6.25 mg/mL according to the compatibility ratio, and administered intragastrically for 1 hour after the operation, the three doses were as high as high. The low values are 1 mg/kg, 5 mg/kg and 25 mg/kg, respectively. Except for the sham operation group, the remaining groups were pulled out to the bifurcation of the internal and external carotid arteries 2 h after ischemia, causing reperfusion injury. The animals were sacrificed at 12 h and 24 h for relevant indicators.

Neurobehavioral Bederson's functional evaluation

Neurobehavioral observations were performed prior to sacrifice, as described by Bederson et al. (Bederson JB, Pitts LH, Tsuji M, et al., Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. Stroke 1986 17:472-476), take the rat tail off the ground about 1 foot, observe the condition of the two forelimbs; place the rats on the level ground, push the shoulders, observe the difference of the resistance on both sides; the rats are placed on the ground, observe Its walking situation. Using a five-point scale (0-4 points), the higher the score, the more serious the neurobehavioral damage.

The neurological function scores are shown in Table 3. The data of each group were compared using the rank sum test, and Ρ<0.05 was considered statistically significant.

Table 3 Effect of the composition of the present invention on neurological function scores of rats after cerebral ischemia-reperfusion. Grouping U quantity (mg-kg ^-1 ) neurological function score

12h 24h

Sham group - 0 0

Model group - 2.62 ± 0.53 2.83 ± 0.65 ^## positive control group 1.30 ± 0.52 1.57 ± 0.46

Administration group 1 1.86±0.67 2.07±0.75*

5 1.52±0.37 1.58±0.59

25 1.24±0.49 1.44±0.65

Data are expressed as mean ± standard deviation and analyzed by one-way ANOVA. Relative to the sham operation group: P<0.01, #P<0.05; relative to the model group: *P<0.05, **P<0.01. Conclusion: The model group was statistically different from the sham operation group, indicating successful modeling (Ρ<0.01), positive control ginkgo biloba extract group (4 mg/kg, ig) and the large, medium and small dose groups of the composition of the present invention. (1, 5mg/kg, 25mg/kg) can significantly improve the neurological symptoms (Ρ<0.05, P<0.01) at 12h and 24h after focal cerebral ischemia and reperfusion in animals, and showed a dose-dependent relationship. .

After functional scoring, the rats were decapitated and the brain tissue was quickly placed in a refrigerator at -20 °C. After lOmin, the room temperature was set. The olfactory bulb, cerebellum and low brain stem were removed and cut into 2 mm crowns according to the brain localization map. Six brains were continuously coronal sections. Then quickly place the brain slices in 5ml containing 2% TTC (triphenyltetrazolium chloride;) and

In a 0.1 ml solution, incubate at 37 ° C for 30 min in the dark, and flip the brain slices once every 5 min. After TTC staining, the normal tissue was rose red, and the infarcted tissue was unstained and white. Each group of brain slices was arranged neatly, photographed and saved, and processed by image analysis system software and counted. The infarct area of each brain slice was calculated, multiplied by the thickness of each brain slice by 2 mm, and the infarct area of each brain slice of each animal was multiplied by The thickness is added, which is the volume of cerebral infarction. The effect of the composition of the present invention on the volume of cerebral infarction after cerebral ischemia and reperfusion is shown in Table 4.

Table 4 The effect of the composition of the present invention on the volume of cerebral infarction after cerebral ischemia and reperfusion. Grouping amount (mg-kg ^-1 ) cerebral infarction volume (mm ³ )

12h 24h

Sham group - 0 0

Model group - 229.35 ± 43.2 248.76 ± 50.23

Positive control group 114.36±54.39** 126.47±38.12**

Administration group 1 168.22±37.20** 179.43±46.73**

5 117.25±42.19** 135.46±42.19**

25 102.38±52.08** 109.53±48.55**

Data are expressed as mean ± standard deviation and analyzed by one-way ANOVA. For sham operation group ^## P <0.01, #P<0.05; for model group *P<0.05, **P<0.01.

Conclusion: As shown in the table, after cerebral ischemia-reperfusion injury in rats, no cerebral infarction occurred in the sham operation group, and the infarct area appeared in the model group. The administration group (1, 5 mg/kg and 25 mg/kg) significantly reduced the volume of cerebral infarction.

Evaluation of brain edema improvement

Cerebral edema is an important pathological process in the early stage of cerebral ischemia. Evaluation of the effect of drugs on cerebral edema is an important indicator for evaluating the role of drug prevention and treatment. The brain tissue water content and brain edema rate are used for evaluation.

Specific test indicators:

Brain tissue water content (%)

Immediately after taking the brain, weigh the wet weight of the ischemic side, the left cerebral hemisphere. Then, the left cerebral hemisphere was baked in an oven at 100 ° C for 24 hours, and then the dry weight was weighed to calculate the water content of the brain tissue. Brain tissue water content (%) = (brain wet Heavy-brain dry weight) Wet weight x 100%.

The effect of the composition of the present invention on the water content of brain tissue of rats after cerebral ischemia-reperfusion is shown in Table 5.

Table 5 Effect of the composition of the present invention on brain water content of rats after cerebral ischemia-reperfusion (±, _n = 7) Grouped U amount (mg-kg ^-1 ) Brain tissue water sprinkling L (100%)

12h 24h

Sham operation group - 78.34±1.15 78.54±0.86

Model group - 81.12±0.98 ^# 81.50±0.74

Positive control group 79.02±0.77** 79.44±0.78** Drug administration group 1 80.18±0.83* 80.73±0.93

5 79.47±0.73** 79.82±0.73**

25 78.86±0.9 wide 78.86±0.9 wide

Conclusion: Cerebral edema rate was statistically different between the model group and the control group at 12h and 24h after cerebral ischemia reperfusion (P < 0.05). The positive control group and the composition of the present invention were compared with the model group. Statistical differences (P < 0.05, P < 0.01) indicate that the composition of the present invention can significantly improve brain edema in rats 12 and 24 hours after cerebral ischemia.

In summary, from the improvement of neurological function score and cerebral edema, the decrease of cerebral infarction volume indicates that the large, medium and small dose groups (1, 5 mg/kg and 25 mg/kg) of the composition of the present invention have obvious protection. The role of injury after cerebral ischemia and reperfusion in rats. Example 3 Experimental study on the protective effect of the composition of the present invention on senile dementia (AD) animals

Incubation of β-amyloid (Αβ _1-40 )

Dilute Αβ^ο to 2 ug/ul with sterile saline and incubate for 1 week at 37 °C to make it condensed Αβι_ ₄₀ °

Establishment of rat dementia model induced by Αβ^ο

250-270 g of healthy male AD rats were anesthetized and fixed on a rat stereotaxic instrument. The skin was routinely sterilized, the skin was cut, the anterior sputum was exposed, and a small syringe was used to the left hippocampus of the rat (with a Bregma point of 0). , AP-3.0mm, ML2.0mm, DV-2.9mm) Slowly inject the condensed state Αβ^ 5μ1, leave the needle for 5min to make the Αβ fully diffuse, then slowly withdraw the needle and suture the incision. The sham operation group only performed preoperative anesthesia, cut the skin, and sputum before exposure, without injection Αβ^^

Experimental grouping and administration

Thirty-two healthy male SD rats of 250-270 g were randomly divided into a sham operation group, a Αβ model group, a Ginkgo biloba leaf extract positive control group, a composition administration group of the present invention 1, 2, 3, and 5 rats in each group. Positive control group administration method: Take 1 tablet (40 mg) of Ginkgo biloba extract tablets, dissolve it in 4 ml of normal saline, prepare 1 mg/mL, and give 1 mL of SD rats by gavage, and the dosage is 4 mg/kg. Administration method of the administration group: Take ginseng total saponins separately: Berberine: Geniposide is formulated into 62.5 mg/mL with physiological saline at a weight ratio of 3:2:0.5, and administered with 2 mL, 1 mL, and 0.5 mL, respectively, in the administration group 1, 2, 3, ie, the dose is 500. Mg/kg, 250 mg/kg and 125 mg/kg. After the model was completed, the Αβ model group was filled with an equal volume of normal saline, and the other groups were filled with the corresponding drugs.

Morris Water Maze Test

Water maze test was started 30 days after surgery. The water maze is a pool with a diameter of 90cm and a height of 40cm. The water depth is 30cm. Four water inlet points are marked on the wall of the pool. The water is divided into four quadrants. A plexiglass circular platform is placed in the center of one of the quadrants. The platform is 2cm below the water surface. The water temperature (24 ± 2) ° C, covered with opaque thick and even white plastic foam, the reference material around the pool remains unchanged. The tests include:

1) Positioning navigation test: It is used to measure the ability of rats to acquire and learn from water fans. For 5 days, two time periods in the afternoon and the afternoon, four times in each time period, the rats were placed into the water from four different marking points, and the time required to find the platform within 2 minutes (evacuation latency) was recorded. If the platform fails to find the platform within 2 minutes after entering the water, place it on the platform for 10 s and escape the incubation period for 2 min. Each training interval is 60s.

2) Space exploration test: It is used to measure the ability of the rats to maintain the platform space position memory after learning to find the platform. After the end of the navigation test, the platform was removed on the sixth day, and a water inlet point was selected to place the rat face into the water, recording the number of times across the original platform within 2 minutes.

One-Way ANOVA was used, and P < 0.05 was considered statistically significant. The results of the Morris water maze test are shown in Table 6.

Table 6 Average escape latency in positioning navigation tests (s, ±sd, n=5)

Day 1 Day 2 Day 3 Day 4 Day Sham Operation Group 74.89±21.30 38.10±14.80 22.60±9.20 21.02±8.73 17.80±7.71 Αβ Model Group 77.80±22.40 59.88±15.50 47.2±11.80" 48.11±11.52' 46.15±15.00

## ## Positive control group 75.30±22.31 40.10±15.20 23.01±9.30, # ^ff # ^ff 21.04±8.75 17.90±7.80

## ## ## Administration group 1 75.10±22.10 39.40±15.0 22.8±9.32 21.30±8.80 17.90±7.80

## ## ## Drug administration group 2 75.20±23.00 39.43±15.30 23.10±9.35 21.50±8.90 17.95±7.85

## ## ## Administration group 3 76.30±23.50 39.90±15.50 23.70±9.45 22.10±9.03 18.30±8.05 Note: P < 0.01, # P < 0.05, ## P < 0.01

Table 7. Results of space exploration experiments in Morris water maze rats (^± ^, n=5) Groups LI quantity (mg/kg) Latency (S) Number of positions through the original platform Sham operation group 10.02±5.38 3.68±1.58

Model group 28.36±11.54 1.69±1.05

Positive control group 4. 15.76±9.79 2.87±1.12 ^#

Drug administration group 1 500 13.34±9.48 3.01±1.36*

Administration group 2 250 15.98±10.02** 2.96±1.09*

Administration group 3 125 17.78±9.96* 2.78±1.12

Note: P < 0.05, P < 0.01 As can be seen from Table 6, the rats did not change significantly in the first 2 days after administration of the composition of the present invention. From the third day, the model group was compared with the sham operation group: "Ρ < 0.01, indicating successful modeling; the administration group and Αβ Comparison of the model groups: ^# Ρ < 0.05, Ρ < 0.01 , which indicates that the administration groups 1, 2, and 3 can significantly shorten the average escape latency from the last three days. Table 7 shows that the drug-administered group can significantly shorten the space exploration of rats. The incubation period increases the number of positions across the original platform, so the composition of the present invention has a significant protective effect on the rat AD model induced by 0 ₁ _ ₄₍₎ . Alumina-induced mouse dementia model and administration method

The experimental animals were selected from 22-month-old Kunming mice, male and female, and randomly divided into 6 groups: normal control group, aluminum trichloride model group, aluminum chloride plus positive control ginkgo biloba extract tablets, aluminum chloride. The compositions of the present invention were administered to groups 1, 2, and 3. The positive control group was administered by a method: 1 tablet (40 mg) of Ginkgo biloba extract was dissolved in 4 ml of physiological saline, and was formulated into 1 mg/mL, and SD rats were intragastrically administered with lmL, and the dose was 4 mg/kg. Preparation method of administration group: Take ginseng total saponin: berberine: geniposide is prepared according to the weight ratio of 3:2:0.5, 62.5 mg/mL with physiological saline, and the administration group 1, 2, 3 respectively 2 mL, 1 mL, and 0.5 mL were administered, ie, doses of 500 mg/kg, 250 mg/kg, and 125 mg/kg. The mice in the aluminum trichloride model group were intragastrically administered with lOmg/ml aqueous solution of aluminum chloride (produced by Shanghai Jinshan Chemical Plant, batch number 100093) at 200 mg/kg body weight/day for 90 days. The ginkgo biloba extract tablets and the composition of the present invention were administered to the positive control group and the administration group while the dementia model was established, and administered by gavage for 90 consecutive days. The normal control group was given the same dose of physiological saline.

Platform test

TT-2 mouse platform automatic control instrument (developed by the Institute of Materia Medica, Chinese Academy of Medical Sciences). The jumping table reflection box is a 90x 18x60cm ³ plexiglass box, which is separated into five by a transparent plate. The bottom of the box is a copper grid, and the voltage is 36V. Each built-in rubber pad with a height of 3.5cm and a diameter of 3.5cm is used as a mouse to avoid electric shock. Safe area. Before the experiment, 3 angels were familiar with the laboratory environment, and the room was kept quiet at a temperature of 25 °C. During the experiment, the mice were placed in a reflective box for 5 min, and then the current was passed at 0.25 mA. The mice were placed on a platform. The mice had the habit of jumping from a height. If they jumped, they were shocked and generated memories. . When the platform has a latency of more than 180 s during training, the number of times the mouse jumped off the platform within 5 minutes (the number of errors) was recorded, and the mice were divided into 6 groups based on the number of errors (normal control group, model group, positive control). Ginkgo biloba extract tablets group, administration group 1, 2, 3, positive control and administration groups 1, 2, 3), 10 mice in each group, the intelligence level of the 6 groups of mice was close. During the experiment, the number of times the mice jumped from the stage (the number of errors) within 5 minutes was carefully determined in detail, which was used as an observation index for learning and memory consolidation in mice.

One-Way ANOVA analysis was considered statistically significant. The results of the platform test are shown in Table 8. Table 8 Comparison of learning and memory ability changes in mouse platform test (±s, n=10)

Group memory acquisition error number of memory memory consolidation error number of normal control group 1.26 ± 0.73 1.80 ± 0.40 model group 6.78 ± 2.40 * 5.90 ± 1.08 ^# positive control group 3.60 ± 2.00 4.78 ± 0.63 ^# drug group 1 2.87 ± 1.90 ^## 3.50±0.88 administration group 2 3.48±2.05 4.10±0.95 administration group 3 4.00±2.10 4.88±1.02 ^#

P<0.05,, #P<0.05, ##P<0.01

As apparent from Table 8, the model group and the treatment group: ^# Ρ <0.05, described successful modeling; Comparative administration group and the model ^{group: # P <0.05, P <} 0.01, show that compositions of the invention can be significantly reduced in mice The number of errors in the dementia model has a significant improvement in the learning and memory ability of mice. Example 4 Experimental study on the protective effect of the composition of the present invention on Parkinson's animal model

Parkinson animal model establishment and administration method

A mouse model of Parkinson's disease was established by intraperitoneal injection of pesticide paraquat (PQ). A total of 120 male Kunming mice, 7 weeks old, were randomly divided into 6 groups, 20 in each group. Blank control group: once a day, distilled distilled water lOmL / kg body weight, once a week intraperitoneal injection of normal saline 10ml / kg body weight; the other 5 groups once a week intraperitoneal injection of PQ10mg / kg, and once a day administration of colchicine, according to adults Four times the dose, ie 0.2 mg/kg, was administered for 7 weeks. After successful modeling, the positive control group: Metoprolol was administered once a day, according to the dose of 7.5 times of the adult dose, that is, the dose was 125 mg/kg, and the administration was continued until one week after the modeling; 2, 3: The mice were intragastrically administered with 250 mg/kg, 125 mg/kg of the composition of the present invention and 62.5 mg/kg of the low-dose group (the preparation method was the same as in Example 3), and the administration was continued until one week after the modeling. Serve under the same conditions, observe the behavioral changes of mice, and then test the content of monoamine neurotransmitters in the substantia nigra of mice. Dopamine (DA;), norepinephrine (NE), 5-hydroxytryptamine (5) -HT).

Rotating rod experiment

Normal mice have the ability to climb, which requires proper grip and motor coordination. We used a rotating rod test to detect motor dysfunction in mice. The mice were placed on a rotating rod tester (YLS-CJ, Beijing Ji'an Deer Technology Co., Ltd.), and each time, 6 rats were rotated at a rate of 20 revolutions per minute, and the number of times the mice fell within 3 minutes was recorded.

Self-determination

The mice were placed in the activity box of the autonomous activity tester, and 6 mice were simultaneously measured at a time, 1 mouse in each box, and the mice were allowed to acclimate for 3 min before starting the test, when the computer entered the measurement procedure. After that, the number of activities and standing of the mouse within 10 minutes was automatically recorded by the computer.

One-Way ANOVA analysis was considered statistically significant. The behavioral test results are shown in Table 9. Table 9 Behavioral examination W results (x±s, n=20)

Rotating rod within 3min, number of horizontal activities, number of standing activities, total number of activities (times)

Number of times (times) (times) (times) blank control group 2.4±0.6 8.4±0.8 76.3±14.5 88.6±15.8

PQ model group 3.5±0.6** 26.8±12.1** 29.6±12.3** Positive control group 3.4±0.7 7.5±0.8 ^## 56.4±10.2 66.4±13.6 ^## administration group 1 3.5±0.4 ^## 6.8±0.7 ^{# #} 58.7 ± 19.6 64.8 ± 17.2 ^## administration group 2 4.8 ± 0.8 # 5.5 ± 0.9 # 63.6 ± 14.3 54.2 ± 11.8 administration group ^{3 5.5 ± 0.7 # 4.0 ± 0.8} 65.5 ± 10.3 # 40.3 ± 15.4 # Note: P < 0.01, #Ρ< 0.05, Ρ< 0.01

As can be seen from Table 9, the number of times the mouse fell off the rotating rod after administration of the composition of the present invention was decreased, and the number of activities was significantly increased. The drug-administered group was compared with the model group: ^# P < 0.05, P < 0.01, indicating that the composition of the present invention can significantly improve the behavioral ability of Parkinson's mice.

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

Table 10 Measurement results of DA, NE, 5 - HT content (; i±s, n=10) Group 5-HT DA NE

(g/g protein) (g/g protein) (g/g protein) blank control group 19.74±10.6 27.96±10.12 23.79±8.6

PQ model group 53.4±7.8** 64.38±9.64* 44.86±10.5** Positive control group 36.53±8.4 ^## 42.28±10.4 ^## 33.24±7.8# Drug administration group 1 35.89±9.5 40.76±8.8 ^## 30.67±8.4 ^# Administration group 2 42.56±6.9 ^# 49.25±12.3 33.12±9.3# Administration group 3 46.88±7.2 50.62±11.2 ^# 39.88±5.4

0.05, P< 0.01, ^ff P<0.05, ^ffff P<0.01

As can be seen from Tables 9, 10, the composition of the present invention can significantly reduce dyskinesia in PQ-induced Parkinson's mice, reduce excessive release of monoamine neurotransmitters, and improve Parkinson's symptoms. Example 5 Experimental study on the protective effect of the composition of the present invention on an animal model of vascular dementia

Animal model establishment

A rat model of bilateral vascular avascular dementia was permanently ligated. After SD rats were intraperitoneally injected with 3.5% chloral hydrate 10ml/kg, the supine position was fixed, the midline incision was made in the neck, and the bilateral common carotid arteries were separated to avoid damage to the cervical sympathetic nerve and vagus nerve. They were permanently ligated with line 1. The wound was sutured after surgery and served in the cage. The sham operation group was to separate the bilateral common carotid arteries, but not to be ligated.

Animal administration method

One month after the operation, the rats were divided into six groups: sham operation group, model group, Ginkgo biloba extract tablets positive control group C4 mg/kg;), the composition administration group 1, 2, 3 (250 mg/kg, 125 mg/kg, 62.5 mg/kg) (Formulation method is the same as in Example 3).

Morris water maze learning memory test

The water maze is a pool with a diameter of 90cm and a height of 40cm. The water depth is 30cm. 4 water inlets are marked on the pool wall. Point, divide it into four quadrants, place a plexiglass circular platform in the center of one quadrant, the platform is 2cm below the water surface, the water temperature is (24±2) °C, covered with opaque thick and uniform white plastic foam, reference around the pool The object remains unchanged. The tests include:

1) Positioning heading experiment: It is used to measure the ability of rats to acquire water memory and learn from memory. Place the platform in the middle of a certain quadrant of the labyrinth, select one of the other three quadrants, and experiment twice a day, each time using a different water inlet point, each training for 1 minute. If the animal has not found the platform within 2 minutes, take the animal to the platform and let it stand for 15 seconds. If the rat finds the platform within 2 minutes, let it stand on the platform for 15 seconds. training. After five days of training, the escape latency and swimming path of each animal were recorded separately.

2) Space search experiment: It is used to measure the ability to maintain the memory of the platform space position after the rats learn to find the platform. After the end of the navigation test, the platform was removed on the sixth day. Each rat was only swam for 2 minutes. The time of the first passage of each rat, ie the incubation period, was recorded with a stopwatch, and each rat was recorded. The number of passes through the platform within 2 minutes, if the incubation period of a rat is shorter and the number of positions passing through the original platform is more, the spatial positioning ability and learning and memory function of the rat are good.

Morris water maze experiments were started 14 days after administration. First, the heading experiment was carried out. Each rat was trained twice a day for a training period of 120 s and continuous training for 5 days. The parameters were recorded. A spatial search experiment was then performed and the platform was removed on the sixth day, recording the latency of the rat and the number of passes through the platform within 2 minutes.

One-way analysis of variance (One-Way ANOVA) analysis, Ρ < 0.05 was considered statistically significant. The results of the water maze experiment are shown in Tables 11, 12, 13, and 14.

Table 11 vascular dementia rats positioning heading experiment escape latency (± s) group dose animal escape latency (S)

(mg/kg) number (only) third day fourth day fifth day sham operation group 12 13.38±5.88 12.04±10.26 9.23±5.32 model group 11 60.24±20.12 ^## 42.32±19.56 ^## 34.42±17.86 ^## positive Control

4 11 20.32±14.24** 17.63±10.25** 14.68±11.54"" group

Administration group 1 250 12 18.76±12.45** 15.56±8.24 13.96±8.71 Administration group 2 125 11 24.22±15.68" 21.25±13.78* 21.21±11.37 ^#药组3 62.5 11 35.79±23.12 ^# 26.38±12.39 24.42± 9.76* Note: P < 0.05, P < 0.01, P < 0.01

As can be seen from Table 11, the model group compared with the sham operation group, ^# P < 0.05, P < 0.01, indicating that the modeling was successful, and the drug-administered group was compared with the model group, *P < 0.05, **P < 0.01, indicating the combination of the present invention. The substance can significantly reduce the escape latency of vascular dementia rats. Table 12 vascular dementia rats positioning heading experimental run (±

Dosage number of animals run (cm)

Group

(mg/kg) (only) third day fourth day fifth day false surgery

12 385.64±214.32 298.79±203.56 212.54±132.74 Group

Model group 11 1647.76±688.43 ^## 1018.04±463.74 ^## 921.64±366.54 ^##正对

4 11 543.22±216.87 356.28±144.84* 346.43±235.21* Photo group

Administration group 1 250 12 606.42±341.64** 394.29±243.66* 332.96±164.82** Administration group 2 125 11 787.21±463.50* 406.78±203.54* Administration group 3 62.5 11 899.54±532.04* 754.74±448.32 592.16±243.12 * Note: P<0.05, P<0.01, P<0.01

There can be seen in Table 12, the model group and the sham ^{group, # P <0.05, P <} 0.01, described successful modeling, comparing administration group and model group, * P <0.05, ** P <0.01, combination of the present invention described The material can significantly reduce the run of the vascular dementia rats in the water maze experiment.

Table 13 Results of space exploration experiments in rats with vascular dementia (±

Number of animals

Latency (s) number of times past the original platform

Sham operation group 12 11.37±5.29 3.68±1.65 Model group 12 28.06±13.57 1.35±1.23 Positive control group 4 12 15.76±8.69 2.88±1.36* Administration group 1 250 12 14.98±9.θΓ ^# 3.00±1.24 ^#药物组2 125 12 17.04 ± 7.32 2.76 ± 1.38 # administered group 3 62.5 12 17.58 ± 10.62 * 2.65 ± 1.44 NOTE: Ρ <0.05, Ρ <0.01

As can be seen from Table 13, the administration group was compared with the model group, *Ρ<0.05, **Ρ<0.01, indicating that the latency of the space exploration experiment after the administration of the composition of the present invention was significantly shorter than that of the model group, and the number of the original platform was significantly shorter. increase.

Table 14 Effect of Compositions of the Invention on the Latent Period Experiment of Looking for Platform in VD Rats

( ±s, n=8)

Looking for platform latency (s)

Day 1 Day 2 Day 3 Day 4 Day 5 Normal group 35.90±24.36 25.90±25.76 23.54±20.12 15.9±9.86 10.8±6.87 Sham operation group 38.60±22.78 28.60±31.42 26.38±20.56 17.6±11.35 15.3±10.34 Model group 67.03±32.74* 62.6±39.88** 56.5±27.58** 42.6±23.08** 38.6±24.64** Ding, ## Administration group 2 37.36±28.32 29.46±21.26 ^ffff 27.14±23.29 ^ffff 20.7±16.72 ^ffff 18.2 ±10.72 Note: *P<0.05, **P<0.01, P<0.01

As can be seen from Table 14, the administration group was compared with the model group, ^## P<0.01, indicating that the rats were administered the composition of the present invention. It can shorten the time to find the platform.

From the above experimental results, it can be seen that the administration of the composition of the present invention can significantly improve the learning and memory ability of the vascular dementia rats.

Dark test

The rats were placed in a bright room and allowed to move freely for 3 min in the box. Those who do not enter the dark room within 3 minutes are excluded (not energized when preselected). After lh, formal training was conducted to record the time required for the rats to enter the dark room to receive electric shock as the incubation period, and the number of electric shocks (the number of errors) within 5 minutes was recorded as the academic achievement; after 24 hours, the above process was repeated, and the latency and the number of errors were recorded as memories. Results. The voltage is 36 V and the latency indicator is in seconds (S).

One-way analysis of variance (One-Way ANOVA) analysis, Ρ < 0.05 was considered statistically significant. The results of the darkness test are shown in Table 15.

Table 15 Results of darkness test in rats with vascular dementia (±s, n=10) group learning training memory test

Incubation period (S) error number latency (S) error number sham operation group 28.69±9.74 1.90±0.86 212.32±19.56 1.52±0.46 model group 49.38±7.32** 7.62±1.78** 87.96±18.64** 7.20±2.18** Medicine group 1 30.46±5.28 3.98±1.24 ^## 146.94±22.40 2.70±1.87 ^## Note: P < 0.01, P < 0.01

As can be seen from Table 15, the administration group 1 was compared with the model group ^## P < 0.01, indicating that the composition of the present invention can significantly shorten the incubation period of the learning training and the memory test, and reduce the number of errors, and it can be seen that the composition of the present invention can improve vascular dementia. Rat's ability to learn and remember.

Conclusion: The composition of the present invention can significantly improve the learning and memory ability of the rats with bilateral common carotid vascular dementia by permanent ligation, and thus has a protective effect on rats with vascular dementia. Example 6 Experimental study on the protective effect of the composition of the present invention on stress and depression animals

Animals were incubated in a quiet, warm environment for 1 week before the experiment. All the animals were kept in a single cage except for the control group. The natural light, well ventilated, and free access to water.

Chronic mild unpredictable stress (CUMS) in Wistar rats. Depression model: 2h behavior limit, swimming in 4°C ice water for 5min, water stop for 24h, food 24h, tail lmin, light and dark upside down 24h, electric shock foot Nine kinds of stimuli, such as 5s (30V voltage), shaking 5min (160Hz), and 40°C environment for 5min, were randomly arranged within 18 days, one per day, so that the animals could not anticipate the occurrence of irritation. Wistar rats were randomly divided into normal control group, depression model group, fluoxetine positive control group, drug administration group 1, 2, 3 (250 mg/kg group, 125 mg/kg group, 62.5 mg/kg group). Same as Example 3) for 21 days, each group of animals was decapitated after 24 hours of the last administration.

Experimental index determination

Behavioral Determination - Morris Water Maze Test

The training and testing of spatial memory was carried out in accordance with the Morris water Maze method. The water temperature is kept at around 23 °C. The water is dyed white to conceal the platform. The surrounding environmental conditions remained quiet and stable during the experiment.

The experiment is divided into training period and testing period, and l-4d is the training period. During the training, the rats were placed in the water against the platform, and the escape latency of the rats was recorded, that is, the time required from the time the rats entered the water to find the underwater hidden platform. If the platform was not found within 60s, the rats were guided to find the platform. . The escape latency is measured in 60s. The rats are found or guided to the platform for 20s, then dried and returned to the cage. Each rat's water entry position is changed clockwise, with each training interval being 30 s. The next day's training is one quadrant into the water than the first day, and so on. The 5th and 7th are the test period, and the platform is removed during the test. Both tests recorded the platform-to-time ratio of the rats, which is the ratio of the time in which the rats in the quadrant of the original platform traveled within 60 seconds of the total time (60s).

One-way analysis of variance (One-Way ANOVA) analysis, Ρ < 0.05 was considered statistically significant. The experimental results are shown in Table 16.

Table 16 Average escape latency of each group of rats during training (±sd, n=12)

Average escape latency (s)

Normal control group 22.1+±4.3

Depression model group 34.9±4.8

Fluoxetine positive control group 26.5±3.7 ^#

Drug administration group 1 26.2±4.5 ^#

Administration group 2 26.7±6.1

Administration group 3 27.5±3.2 ^#

Note: P < 0.01, ^# P < 0.05

As can be seen from Table 16, the drug-administered group was compared with the model group: ^# P < 0.05, indicating that the rat's escape latency was significantly shortened after administration of the composition of the present invention, that is, the composition of the present invention can improve the learning and memory ability of the stress-induced depression rats. .

Determination of biochemical indicators - determination of monoamine oxidase (MAO), glutathione S-transferase (GST), gamma glutamyl transferase (γ-GT)

After the last treatment of Wistar rat animals, the rats were decapitated and the cerebral cortex of the rats was rapidly separated on an ice tray, and the lower brain stem was removed. After weighing, 0.86% pre-cooled physiological saline 1:9 was homogenized in an ice water bath to prepare a 10% brain homogenate. After each sample was dispensed in several portions, the refrigerator was stored in a low temperature freezer. The prepared homogenate was centrifuged at 3000 r/min, and centrifuged for 15 minutes to obtain a homogenate supernatant, and the enzyme activities of MAO, GST, and γ-GT were respectively determined by a chromogenic substrate method according to the method described in the kit. The results are shown in Table 17.

Table 17 Results of biochemical indicators of rat brain homogenate (;^ ±sd, n=12)

MAO activity GST activity γ-GT activity group

μ/g protein μ/g protein μ/g protein normal control group 20.35 ±9.23 18.79±4.78 87.66±20.45 depression model group 35.24±7.45 33.76±6.33** 59.64±17.21 "fluoxetine positive control group 21. 87±9.64 ^M 20.04±8.34 Ding 76.13±22.75 Pu-administered group 1 22.19±7.73 Pu 22.15±5.82 Pu 79.65±18.39 ^## administration group 2 24.38±9.67 Pu 25.83±5.46 Pu 74.37±20.15 ^#药物组3 26.02±7.58 ^# 27.01±3.98 ^# 72.97±23.64 ^#

0.01, # P < 0.05, ## P < 0.01 As can be seen from Table 17, the drug-administered group was compared with the model group: ^# P < 0.05, P < 0.01, indicating that the composition of the present invention can significantly increase monoamine oxidase (MAO), glutathione S-transferase (GST), γ-glutamyl Transferase (γ-GT) activity.

Conclusion: The composition of the invention has a certain protective effect on chronic stress-induced depression rats. Example 7 Experimental study on the protective effect of the composition of the present invention on bacterial meningitis

Establishment of a bacterial meningitis model

Experimental animal

Because bacterial meningitis has a high incidence in infants and young children and is a serious acute infectious disease in the neonatal period, the experimental animals of bacterial meningitis use 11-day-old SD newborn rats.

Pathogenic bacteria: GBSIII strains were purchased from China National Institute for the Control of Pharmaceutical and Biological Products.

The dried strains were cultured in serum broth for 24 hours at 37 ° C, and then streaked into blood plates. After 18-24 hours, a single colony was selected into serum broth medium to collect in the middle of logarithmic growth. The OD value corresponding to the adjusted turbid liquid concentration by the turbidimetric method was the same as that of the 0.5 standard turbidity tube (concentration lx l0 ⁸ cfu/ml), and the bacterial liquid concentration was 10 ⁸ cfu/ml. Dilute the bacteria to the desired concentration of 10 ⁴ cfu/ml with physiological saline.

1) Experimental group SD neonatal rats were anesthetized by intraperitoneal injection of 1.0% sodium pentobarbital (15-25 mg/kg). Hold the lOul puncture needle at the midpoint of the midpoint of the two ear tips of the newborn rats. The 2-4mm depression (cerebellar medullary pool) is inserted vertically. After a clear breakthrough, the clear cerebrospinal fluid (CSF) flows out and the l OulCSF is discarded. . The experimental group was slowly injected with 10 ul of GSBIII suspension. The whole process was completed in 1 and 2 minutes. After surgery, the patient was placed at a room temperature of 25 °C. After resuscitation, the rats were returned to the cage and fed by the mother. After culturing for a certain period of time, the experimental group CSF is extracted and processed into a CSF stock solution, cefotaxime + CSF, the composition of the present invention 500 g / kg + CSF, the composition of the present invention 250 mg / kg + CSF, the present invention Compositions 125 mg/kg + CSF Several different groups of brain infusions.

2) Fresh SD rats were randomly divided into: blank control group, model group, positive control group, and drug group 1, 2, 3. Anesthesia was administered intraperitoneally with 1.0% sodium pentobarbital (15-25 mg/kg). Hold the lOul puncture needle at the midpoint of the midpoint of the two ear tips of the newborn rats. The 2-4mm depression (cerebellar medullary pool) is inserted vertically. After a clear breakthrough, the clear cerebrospinal fluid (CSF) flows out and the lOulCSF is taken out. The same amount of CSF stock solution, cefotaxime + CSF, 500 mg/kg + CSF of the present invention (administered group 1), and 250 mg/kg + CSF of the present invention were administered to each group (administration group 2) The composition of the invention is 125 mg/kg + CSF (administration group 3). Postoperative supine, set at room temperature 25 °C, put back into the cage after resuscitation, continue feeding by the mother.

Clinical symptom observation

After inoculation of the bacteria, clinical observations include: presence or absence of cranial and limb skin blemishes, difficulty breathing, unresponsiveness, decreased exercise, and convulsions, as well as the time at which symptoms first appear. Neonatal rats were evaluated 24 hours after inoculation with Loeffler's neurobehavioral 5-point scale. 5 points: When grabbing the back, you can move normally, turn over within 5s; 4 points, reduce spontaneous movement, turn over within 5s; 3 points: 〉5s turn over; 2 points: Can't turn over; 1 point: Can't move. For the occurrence of convulsions, the degree of convulsion was evaluated according to Racine's sputum grading criteria for SD rats. The standard is divided into 6 levels, level 0: no convulsions; level I: facial palsy; level II: on the basis of level I, nodular nod; level III: on the basis of level II with forelimb clonic; IV Level: There is hind leg standing force on the basis of Level III; Class V: There is a fall on the basis of Grade IV.

The experimental results are shown in Tables 18 and 19.

Table 18 Comparison of clinical manifestations of neonatal rats vaccinated with different fluids

Bleeding, difficulty breathing, movement disorder

Blank control group 10 0 0 0 0 model group 10 9 9 7 6 positive control group 10 2 1 2 1 administration group 1 10 2 2 2 2 administration group 2 10 3 4 2 3 administration group 3 10 3 5 3 3 As can be seen from Table 18, the clinical symptoms of the rats of the present invention were significantly alleviated after administration of the composition of the present invention.

Table 19 Inoculation of different fluids 24h neurobehavioral score comparison group n score (±s)

Blank control group 10 5±0

Model group 10 2.87±1.09

Positive control group 10 3.99±0.87 ^##

Administration group 1 10 4.27±1.12 ^##

Drug administration group 2 10 3.79±1.04 ^##

Administration group 3 10 3.43±1.36 ^#

Note: P < 0.01, # P < 0.05, P < 0.01

As can be seen from Table 19, One-Way ANOVA analysis, the model group was compared with the blank control group: "Ρ < 0.01 indicates successful modeling; the drug-administered group compared with the model group: #Ρ<0.05, ^## Ρ <0.01, indicating that the neurological behavior of the drug-administered group was significantly improved compared with the model group, and it can be seen that the composition of the present invention can significantly improve the neurobehavior of meningitis rats.

Rats that survived after 24 h of inoculation were sacrificed by cerebellar medullary puncture for lOul CSF for WBC (white blood cell calculation) before enumeration. The results are shown in Table 20 below:

Table 20 WBC calculation results

Group n mean (±s) blank control group 5 13.8±1.23

Model group 5 14780.0±5978** positive control group 5 9920±1745 ^#

Administration group 1 5 9054±2346

Drug administration group 2 5 9847±3890

Administration group 3 5 10976±4577 ^#

Note: P < 0.01, ^# P < 0.05, P < 0.01 As can be seen from Table 20, One-Way ANOVA analysis, the model group was compared with the blank control group: "P < 0.01, indicating successful modeling; the drug-administered group compared with the model group: #P<0.05, ^## P < 0.01, indicating that the composition of the present invention can significantly reduce the number of white blood cells in rats and reduce the inflammation in rats with meningitis.

Conclusion: The composition of the present invention has a good antibacterial effect and has a certain improvement effect on the symptoms of bacterial meningitis animals.

Claims

Claims:

A traditional Chinese medicine active ingredient composition comprising geniposide or geniposide, berberine, ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition.

The traditional Chinese medicine active ingredient composition according to claim 1, characterized in that the following parts by weight are included in a total weight of 100 parts: ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition, 1 -99 parts, berberine, 1-99 parts, geniposide or geniposide, 1-99 parts; preferably ginseng total saponins or ginsenoside monomers or ginsenoside monomer compositions, 2-75 parts, berberine , 2-75 parts, geniposide or geniposide, 2-75 parts; more preferably ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition, 3-50 parts, berberine, 3-50 parts , geniposide or geniposide, 3-50 parts.

The traditional Chinese medicine active ingredient composition according to claim 2, wherein the weight ratio of ginseng total saponin or ginsenoside monomer or ginsenoside monomer composition, berberine, geniposide or geniposide is 3 : 2: 2-0.1, preferably 3:2: 2-1, more preferably 3:2:2.

4. A pharmaceutical preparation comprising the composition according to claim 1 as an active ingredient together with any one or more pharmaceutically acceptable carriers.

5. A pharmaceutical preparation according to claim 4 which can be formulated into any of the pharmaceutically acceptable dosage forms.

6. Use of a composition according to claim 1 for the manufacture of a medicament for the treatment or prevention of brain and nerve damage, neurodegenerative encephalopathy, bacterial or viral encephalitis, or depression.

7. The use according to claim 6, wherein the brain and nerve damage is cerebrovascular dementia or ischemic cerebrovascular disease.

8. The use according to claim 7, wherein the ischemic cerebrovascular disease is a transient partial cerebral infarction, cerebral infarction or stroke.

9. The use according to claim 6, wherein the neurodegenerative encephalopathy is Alzheimer's disease or Parkinson's disease.